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Pisetsky, David Stephen

Overview:

Studies in my laboratory concern the immunological properties of DNA as related to two main topics: 1) the induction of anti-DNA responses in systemic lupus erythematosus; and 2) the stimulation of innate immunity by bacterial DNA. These topics are closely linked since we have established novel disease models in which lupus-like illness can be induced in normal mice by bacterial DNA under conditions in which mammalian DNA is inactive. This model has been useful in elucidating mechanisms of DNA antigen drive in autoimmunity.

To pursue these studies, our laboratory conducts investigations in the following areas: 1) specificity of anti-DNA for epitopes on mammalian and bacterial DNA; 2) molecular analysis of murine mononclonal anti-DNA antibodies; 3) histopathological analyses of DNA-immunized mice; 4) in vitro analysis of proliferation, antibody production and cytokine expression in human and murine immune cells; and 5) analysis of DNA binding to cell surface molecules on B cells, T cells and macrophages. Results of these studies have allowed identification of at least two structural motifs that are immunoactive. We are also exploring the impact of chemical modifications of the DNA backbone.

In addition to work on the immunology of DNA, I am also involved in clinical trials related to new immunomodulatory agents in the treatment of rheumatoid arthritis as well as serological markers of disease activity.

The areas of research for which I am known nationally are anti-DNA antibodies, systemic lupus erythematosus and immunological properties of DNA. I have written textbook chapters and reviews on all these subjects.

Positions:

Professor of Medicine

Medicine, Rheumatology and Immunology
School of Medicine

Professor of Immunology

Immunology
School of Medicine

Member of the Duke Cancer Institute

Duke Cancer Institute
School of Medicine

Member of the Duke Human Vaccine Institute

Duke Human Vaccine Institute
School of Medicine

Education:

Ph.D. 1972

Ph.D. — Albert Einstein College of Medicine of Yeshiva University

M.D. 1973

M.D. — Albert Einstein College of Medicine of Yeshiva University

News:

Grants:

Interdisciplinary Training Program in Lung Disease

Administered By
Medicine, Pulmonary, Allergy, and Critical Care Medicine
AwardedBy
National Institutes of Health
Role
Preceptor
Start Date
July 01, 2009
End Date
March 31, 2020

Basic Immunology Training Program

Administered By
Immunology
AwardedBy
National Institutes of Health
Role
Mentor
Start Date
July 01, 2002
End Date
June 30, 2019

Metabolomic Analysis of Paired Serum and Skeletal Muscle in Juvenile Inflammatory Myositis Compared to Controls

Administered By
Pediatrics, Rheumatology
AwardedBy
Cure JM Foundation
Role
Co Investigator
Start Date
February 01, 2017
End Date
January 31, 2019

Training Program in Inflammatory and Immunological Diseases

Administered By
Medicine, Rheumatology and Immunology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
September 30, 1980
End Date
August 31, 2017

Determinants of ANA Expression in Patients with SLE

Administered By
Medicine, Rheumatology and Immunology
AwardedBy
Lupus Research Institute
Role
Principal Investigator
Start Date
July 15, 2015
End Date
April 01, 2017

Obesity, Biomechanics, and Inflammation in Osteoarthritis

Administered By
Orthopaedics
AwardedBy
National Institutes of Health
Role
Collaborator
Start Date
September 30, 2013
End Date
January 31, 2016

IPA - Diane Spencer

Administered By
Medicine, Rheumatology and Immunology
AwardedBy
Durham Veterans Affairs Medical Center
Role
Principal Investigator
Start Date
September 17, 2012
End Date
September 16, 2014

IPA - Nancy Stearns

Administered By
Medicine, Rheumatology and Immunology
AwardedBy
Durham Veterans Affairs Medical Center
Role
Principal Investigator
Start Date
September 17, 2012
End Date
September 16, 2014

Live-Animal Micro-CT System

Administered By
Orthopaedics
AwardedBy
National Institutes of Health
Role
Major User
Start Date
May 15, 2012
End Date
May 14, 2014

Nucleic Acid Binding Polymers as Anti-Inflammatory Agents

Administered By
Surgery, Surgical Sciences
AwardedBy
National Institutes of Health
Role
Co Investigator
Start Date
September 01, 2011
End Date
August 31, 2013

The Role of Sex in Self Antigen Generation in SLE

Administered By
Medicine, Rheumatology and Immunology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
July 17, 2009
End Date
June 30, 2012

Microparticles as a Source of Nuclear Antigens in Systemic Lupus Erythematosus

Administered By
Medicine, Rheumatology and Immunology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
May 22, 2009
End Date
March 31, 2012

IPA - Julie Lynn Gauley

Administered By
Medicine, Rheumatology and Immunology
AwardedBy
Veterans Administration Medical Center
Role
Principal Investigator
Start Date
July 01, 2011
End Date
December 31, 2011

IPA - Julie Gauley

Administered By
Medicine, Rheumatology and Immunology
AwardedBy
Veterans Administration Medical Center
Role
Principal Investigator
Start Date
December 01, 2009
End Date
November 30, 2010

Nitric Oxide, Oxygen and Mechanical Stress in Cartilage

Administered By
Orthopaedics
AwardedBy
National Institutes of Health
Role
Co Investigator
Start Date
August 01, 2004
End Date
February 01, 2010

IPA Charles Reich

Administered By
Medicine, Rheumatology and Immunology
AwardedBy
Veterans Administration Medical Center
Role
Principal Investigator
Start Date
February 01, 2008
End Date
January 31, 2010

IPA Julie Lynn Gauley

Administered By
Medicine, Rheumatology and Immunology
AwardedBy
Veterans Administration Medical Center
Role
Principal Investigator
Start Date
December 01, 2008
End Date
November 30, 2009

Training in Biomolecular and Tissue Engineering

Administered By
Orthopaedics
AwardedBy
National Institutes of Health
Role
Mentor
Start Date
September 20, 2003
End Date
June 30, 2009

IPA - Charles Reich

Administered By
Medicine, Rheumatology and Immunology
AwardedBy
VAMC
Role
Principal Investigator
Start Date
February 01, 2007
End Date
January 31, 2008

The Immunomodulatory Effects of DNA in SLE

Administered By
Medicine, Rheumatology and Immunology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
April 15, 2001
End Date
March 31, 2007

Planning an Early RA Prevention Trial

Administered By
Medicine, Rheumatology and Immunology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
August 01, 2005
End Date
July 31, 2006

Strategies for Evaluating and Treating Early Synovitis

Administered By
Medicine, Rheumatology and Immunology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
September 25, 2003
End Date
June 30, 2006

Mechanical Stress and Prostaglandins in Cartilage

Administered By
Orthopaedics
AwardedBy
National Institutes of Health
Role
Collaborator
Start Date
August 01, 2001
End Date
May 31, 2004

Mechanisms of Anti-DNA Production in Autoimmune Mice

Administered By
Medicine, Rheumatology and Immunology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
September 30, 2000
End Date
September 29, 2001

Specialized center of research (SCOR) in Rheumatoid Arthritis

Administered By
Medicine, Rheumatology and Immunology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
September 30, 1987
End Date
August 31, 2001

Specialized Center Of Research In Rheumatoid Arthritis

Administered By
Medicine, Rheumatology and Immunology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
September 01, 1987
End Date
August 31, 1999

Training Program - Inflammatory And Immunological Diseases

Administered By
Medicine, Rheumatology and Immunology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
September 30, 1980
End Date
June 30, 1999

Intravenous Doxycycline For Rheumatoid Arthritis

Administered By
Medicine, Rheumatology and Immunology
AwardedBy
National Institutes of Health
Role
Co-Principal Investigator
Start Date
September 30, 1994
End Date
August 31, 1998

Specialized Center Of Research In Rheumatoid Arthritis

Administered By
Medicine, Rheumatology and Immunology
AwardedBy
National Institutes of Health
Role
Co-Principal Investigator
Start Date
September 30, 1987
End Date
August 31, 1997

Specialized Center Of Research In Rheumatoid Arthritis

Administered By
Medicine, Rheumatology and Immunology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
September 30, 1987
End Date
August 31, 1997

Identification Of Autoimmune Genes In 1pr Mice

Administered By
Medicine, Rheumatology and Immunology
AwardedBy
National Institutes of Health
Role
Co-Principal Investigator
Start Date
September 30, 1992
End Date
August 31, 1996

Identification Of 'Autoimmune Genes' In 1pr/1pr Mice

Administered By
Medicine, Rheumatology and Immunology
AwardedBy
National Institutes of Health
Role
Co-Principal Investigator
Start Date
September 30, 1992
End Date
July 31, 1996

Molecular And Cellular Mechanisms Of Human Inflammatory

Administered By
Medicine, Rheumatology and Immunology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
September 01, 1990
End Date
August 31, 1992

Molecular And Cellular Mechanisms Of Human Inflammatory D.

Administered By
Medicine, Rheumatology and Immunology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
September 01, 1989
End Date
August 01, 1991

Molecular Mechanisms Of Human Inflammatory Diseases

Administered By
Medicine, Rheumatology and Immunology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
September 01, 1988
End Date
August 01, 1991

Molecular Mechanisms Of Human Inflammatory Disease

Administered By
Medicine, Rheumatology and Immunology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
September 01, 1987
End Date
August 01, 1991

Molecular And Cellular Mechanisms Of Human Inflammatory Di

Administered By
Medicine, Rheumatology and Immunology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
September 01, 1986
End Date
August 01, 1991
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Publications:

Advances in the Treatment of Rheumatoid Arthritis: Costs and Challenges.

Rheumatoid arthritis is the most common form of inflammatory arthritis. Because of advances in therapy, clinical outcomes have improved dramatically and remission is possible for many patients. These advances have come with many challenges, prompting consideration of strategies to improve diagnosis and treatment and implement more cost-effective care.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Advances in the Treatment of Rheumatoid Arthritis: Costs and Challenges." North Carolina medical journal 78.5 (September 2017): 337-340.
PMID
28963273
Source
epmc
Published In
North Carolina Medical Journal
Volume
78
Issue
5
Publish Date
2017
Start Page
337
End Page
340
DOI
10.18043/ncm.78.5.337

EULAR recommendations for disease management: guidance not guidelines.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "EULAR recommendations for disease management: guidance not guidelines." Annals of the rheumatic diseases 76.6 (June 2017): 935-938.
PMID
28465346
Source
epmc
Published In
Annals of the rheumatic diseases
Volume
76
Issue
6
Publish Date
2017
Start Page
935
End Page
938
DOI
10.1136/annrheumdis-2016-211005

Pain management in rheumatology research, training, and practice

© Clinical and Experimental Rheumatology 2017. The Pain Management Task Force of the American College of Rheumatology published a report in 2010 highlighting pain management as a fundamental aspect of clinical practice, training and research. In the interim, the consideration of pain as a focus of attention of rheumatologists and rheumatology health professionals has become even more challenging than in 2010 because of the epidemic of opiate addiction and overdose death. The characterisation of categories of pain by mechanism (e.g., inflammation, joint degeneration, abnormalities of central pain processing) can help guide treatment. However, such categorisation can overlook the overlap of these processes and their interaction to create mixed pain states. Further complicating the assessment of pain, outcome measures in rheumatic disease often assess the degree of pain indirectly while concentrating on the quantification of inflammation. Non-inflammatory pain often persists despite treatment, highlighting the need for alternative analgesic therapies. Recommended therapies include acetaminophen, nonsteroidal anti-inflammatory drugs, and stimulators of the pain inhibitory pathway. Each of these non-opioid therapies has incomplete efficacy and potential toxicities that can limit their utility. Non-pharmacologic therapies can show efficacy that rivals or surpasses pharmacologic therapies in the control of pain and improving function in a variety of rheumatic disorders including chronic low back pain and fibromyalgia. A limitation of the use of these therapies is inadequate training and appreciation of their benefits. Furthermore, the supply of trained practitioners to provide non-pharmacological care and support patient efforts for self-management is often limited. Together, these considerations suggest the importance of a renewed effort to implement task force recommendations.

Authors
Borenstein, DG; Hassett, AL; Pisetsky, DS
MLA Citation
Borenstein, DG, Hassett, AL, and Pisetsky, DS. "Pain management in rheumatology research, training, and practice." Clinical and Experimental Rheumatology 35.5 (January 1, 2017): S2-S7.
Source
scopus
Published In
Clinical and experimental rheumatology
Volume
35
Issue
5
Publish Date
2017
Start Page
S2
End Page
S7

Mechanisms of Chromatin Remodeling and Repurposing During Extracellular Translocation.

Chromatin is a highly conserved molecular structure that provides genetic information to regulate cell function. Comprised of DNA, histones and interacting proteins, chromatin is inherently dynamic and subject to remodeling. While usually conceptualized as an intranuclear event, remodeling can also involve extracellular movement. Indeed, chromatin can translocate entirely from the inside to the outside of the cell during cell death processes that include apoptosis, necrosis, and NETosis. During these processes, DNA and proteins can undergo other changes impacting on their activity. Thus, during apoptosis, DNA can be cleaved, histones can be posttranslationally modified and a nuclear protein called HMGB1 (high mobility group box 1) can undergo redox changes. Outside the cell, chromatin components can display powerful immunological activities. These activities result from the ability of DNA and RNA, once taken up by immune cells, to activate internal nucleic acid sensors; the likely function of these sensors is to recognize nucleic acids from intracellular infection. Depending on redox state, the prototype alarmin HMGB1 can interact with a variety of immune receptors including Toll-like receptors. As such, extracellular chromatin can stimulate inflammation and drive the pathogenesis of immune-mediated diseases; in experimental models in animals, agents that bind chromatin components can block disease. Thus, extracellular chromatin can have far-reaching biological effects involving a form of molecular repurposing.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Mechanisms of Chromatin Remodeling and Repurposing During Extracellular Translocation." Advances in protein chemistry and structural biology 106 (January 2017): 113-137. (Review)
PMID
28057209
Source
epmc
Published In
Advances in Protein Chemistry and Structural Biology
Volume
106
Publish Date
2017
Start Page
113
End Page
137
DOI
10.1016/bs.apcsb.2016.08.003

Tapering biologic and conventional DMARD therapy in rheumatoid arthritis: current evidence and future directions.

Improvements in the control of inflammation in rheumatoid arthritis (RA) by conventional synthetic and biologic disease-modifying antirheumatic drugs (DMARDs) have led to a substantial change in the clinical outcomes of patients during the last 30 years. Current treatment can lead to sustained remission in some patients raising questions about the optimal management strategies in this subgroup of patients. Today, tapering of DMARDs and even their discontinuation appears as an interesting concept for achieving a more tailored and dynamic treatment approach of RA, especially in patients, who achieved full disease control by DMARD treatment. In this review article, current developments of DMARD tapering are discussed. The article provides an overview of existing studies on this topic and addresses new strategies to reach drug-free remission. Furthermore, concepts for defining patients eligible for DMARD tapering are described and potential future strategies in using biomarkers in predicting the risk for disease relapse after initiation of DMARD tapering are addressed. These findings are finally considered in light of the vision to achieve cure as an ultimate goal in patients with RA achieving full control of inflammation.

Authors
Schett, G; Emery, P; Tanaka, Y; Burmester, G; Pisetsky, DS; Naredo, E; Fautrel, B; van Vollenhoven, R
MLA Citation
Schett, G, Emery, P, Tanaka, Y, Burmester, G, Pisetsky, DS, Naredo, E, Fautrel, B, and van Vollenhoven, R. "Tapering biologic and conventional DMARD therapy in rheumatoid arthritis: current evidence and future directions." Annals of the rheumatic diseases 75.8 (August 2016): 1428-1437. (Review)
PMID
27261493
Source
epmc
Published In
Annals of the rheumatic diseases
Volume
75
Issue
8
Publish Date
2016
Start Page
1428
End Page
1437
DOI
10.1136/annrheumdis-2016-209201

The expression of microvesicles in the blood of patients with Graves' disease and its relationship to treatment.

Graves' disease (GD) is an autoimmune disease characterized by the presence of circulating autoantibodies against thyroid-stimulating hormone (TSH) receptor. Despite extensive research, the pathogenic mechanisms remain unclear. Immune responses associated with the disease may lead to cell activation/apoptosis and the release of microvesicles (MVs) into the circulation. MVs can display biological activities which may aggravate GD further. We studied immune mechanisms in GD by investigating the numbers and phenotype of circulating MVs in patients before and after antithyroid therapy with thiamazole.Samples were obtained from 15 patients with GD in the acute phase of hyperthyroidism and following 17-26 months treatment and 14 healthy controls. MVs from platelets, endothelial cells and monocytes exposing inflammation/activation markers (P-selectin, CD40 ligand, E-selectin and HMGB1) and MVs containing nuclear molecules were measured with flow cytometry.Patients had elevated baseline values of MVs (P < 0·001 for all types of MVs), while the levels decreased during thiamazole treatment (P < 0·05 for all types of MVs). The majority of MV populations remained, however, significantly higher in patients after treatment compared to levels in controls.GD patients have elevated levels of MVs that carry molecules with potential biological activities. MVs are significantly reduced after antithyroid treatment with thiamazole but still higher compared to levels in healthy controls. Assessment of MV levels and pattern may therefore provide additional information on underlying immune disturbances not obtained by measurements of hormone levels alone.

Authors
Mobarrez, F; Abraham-Nordling, M; Aguilera-Gatica, K; Friberg, I; Antovic, A; Pisetsky, DS; Jörneskog, G; Wallen, H
MLA Citation
Mobarrez, F, Abraham-Nordling, M, Aguilera-Gatica, K, Friberg, I, Antovic, A, Pisetsky, DS, Jörneskog, G, and Wallen, H. "The expression of microvesicles in the blood of patients with Graves' disease and its relationship to treatment." Clinical endocrinology 84.5 (May 2016): 729-735.
PMID
26252432
Source
epmc
Published In
Clinical Endocrinology
Volume
84
Issue
5
Publish Date
2016
Start Page
729
End Page
735
DOI
10.1111/cen.12872

The Alarmin Properties of DNA and DNA-associated Nuclear Proteins.

The communication of cell injury and death is a critical element in host defense. Although immune cells can serve this function by elaborating cytokines and chemokines, somatic cells can repurpose nuclear macromolecules to function as damage-associated molecular patterns (DAMPs) or alarmins to exert similar activity. Among these molecules, DNA, high-mobility group box-1, and histone proteins can all act as DAMPs once they are in an extracellular location. This review describes current information on the role of the nuclear DAMPs, their translocation to the outside of cells, and pathways of activation after uptake into the inside of immune cells.MEDLINE and PubMed databases were searched for citations (1990-2016) in English related to the following terms: DAMPs, high-mobility group box-1, DNA, histones, cell death, danger, and immune activation. Selected articles with the most relevant studies were included for a more detailed consideration.Although nuclear molecules have important structural and genetic regulatory roles inside the cell nucleus, when released into the extracellular space during cell death, these molecules can acquire immune activity and serve as alarmins or DAMPs. Although apoptosis is generally considered the source of extracellular nuclear material, other cell death pathways such as necroptosis, NETosis, and pyroptosis can contribute to the release of nuclear molecules. Importantly, the release of nuclear DAMPs occurs with both soluble and particulate forms of these molecules. The activity of nuclear molecules may depend on posttranslational modifications, redox changes, and the binding of other molecules. Once in an extracellular location, nuclear DAMPs can engage the same pattern recognition receptors as do pathogen-associated molecular patterns. These interactions can activate immune cells and lead to cytokine and chemokine production. Among these receptors, internal receptors for DNA are key to the response to this molecule; the likely function of these internal sensors is the recognition of DNA from intracellular infection by bacteria or viruses. Activation of these receptors requires translocation of extracellular DNA into specialized compartments. In addition to nuclear DNA, mitochondrial DNA can also serve as a DAMP.The communication of cell injury and death is a critical element in host defense and involves the repurposing of nuclear molecules as immune triggers. As such, the presence of extracellular nuclear material can serve as novel biomarkers for conditions involving cell injury and death. Targeting of these molecules may also represent an important new approach to therapy.

Authors
Magna, M; Pisetsky, DS
MLA Citation
Magna, M, and Pisetsky, DS. "The Alarmin Properties of DNA and DNA-associated Nuclear Proteins." Clinical therapeutics 38.5 (May 2016): 1029-1041. (Review)
PMID
27021604
Source
epmc
Published In
Clinical Therapeutics
Volume
38
Issue
5
Publish Date
2016
Start Page
1029
End Page
1041
DOI
10.1016/j.clinthera.2016.02.029

The role of monogamous bivalency and Fc interactions in the binding of anti-DNA antibodies to DNA antigen.

Antibodies to DNA (anti-DNA) are the serological hallmark of systemic lupus erythematosus. These antibodies can bind DNA avidly by monogamous bivalency, a mechanism which requires the interaction of both Fab combining regions with antigenic determinants on the same polynucleotide. To explore further this mechanism, we tested Fab and F(ab')2 fragments prepared from IgG from patient plasmas in an ELISA with native DNA antigen, detecting antibody with a peroxidase conjugated anti-Fab reagent. These studies showed that Fab fragments, which can only bind monovalently, had negligible activity. Although bivalent F(ab')2 fragments would be predicted to bind DNA, these fragments also showed poor anti-DNA activity. Control studies showed that the fragments retained antibody activity to tetanus toxoid and an EBV antigen preparation. Together, these findings suggest that anti-DNA avidity depends on monogamous bivalency, with the antibody Fc portion also influencing DNA binding, in a mechanism which can be termed Fc-dependent monogamous bivalency.

Authors
Stearns, NA; Pisetsky, DS
MLA Citation
Stearns, NA, and Pisetsky, DS. "The role of monogamous bivalency and Fc interactions in the binding of anti-DNA antibodies to DNA antigen." Clinical immunology (Orlando, Fla.) 166-167 (May 2016): 38-47.
PMID
27083935
Source
epmc
Published In
Clinical Immunology
Volume
166-167
Publish Date
2016
Start Page
38
End Page
47
DOI
10.1016/j.clim.2016.04.004

Anti-DNA antibodies--quintessential biomarkers of SLE.

Antibodies that recognize and bind to DNA (anti-DNA antibodies) are serological hallmarks of systemic lupus erythematosus (SLE) and key markers for diagnosis and disease activity. In addition to common use in the clinic, anti-DNA antibody testing now also determines eligibility for clinical trials, raising important questions about the nature of the antibody-antigen interaction. At present, no 'gold standard' for serological assessment exists, and anti-DNA antibody binding can be measured with a variety of assay formats, which differ in the nature of the DNA substrates and in the conditions for binding and detection of antibodies. A mechanism called monogamous bivalency--in which high avidity results from simultaneous interaction of IgG Fab sites with a single polynucleotide chain--determines anti-DNA antibody binding; this mechanism might affect antibody detection in different assay formats. Although anti-DNA antibodies can promote pathogenesis by depositing in the kidney or driving cytokine production, they are not all alike, pathologically, and anti-DNA antibody expression does not necessarily correlate with active disease. Levels of anti-DNA antibodies in patients with SLE can vary over time, distinguishing anti-DNA antibodies from other pathogenic antinuclear antibodies. Elucidation of the binding specificities and the pathogenic roles of anti-DNA antibodies in SLE should enable improvements in the design of informative assays for both clinical and research purposes.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Anti-DNA antibodies--quintessential biomarkers of SLE." Nature reviews. Rheumatology 12.2 (February 2016): 102-110. (Review)
PMID
26581343
Source
epmc
Published In
Nature Reviews Rheumatology
Volume
12
Issue
2
Publish Date
2016
Start Page
102
End Page
110
DOI
10.1038/nrrheum.2015.151

The role of mitochondria in immune-mediated disease: the dangers of a split personality.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "The role of mitochondria in immune-mediated disease: the dangers of a split personality." Arthritis research & therapy 18 (January 2016): 169-.
PMID
27424174
Source
epmc
Published In
Arthritis Research and Therapy
Volume
18
Publish Date
2016
Start Page
169
DOI
10.1186/s13075-016-1063-5

The Use of Poly-L-Lysine as a Capture Agent to Enhance the Detection of Antinuclear Antibodies by ELISA.

Antibodies to nuclear antigens (antinuclear antibodies or ANAs) are the serological hallmark of systemic lupus erythematosus (SLE). These antibodies bind diverse nuclear antigens that include DNA, histones and non-histone proteins as well as complexes of proteins with DNA and RNA. Because of the frequency of ANA expression in SLE, testing is an important component of clinical evaluation as well as determination of eligibility for clinical trials or utilization of certain therapies. Immunofluorescence assays have been commonly used for this purpose although this approach can be limited by issues of throughput, variability and difficulty in determining positivity. ELISA and multiplex assays are also useful approaches although these assays may give an incomplete picture of antibodies present. To develop a sensitive and quantitative ANA assay, we have explored an ELISA platform in which plates are pre-coated with a positively charged nucleic acid binding polymer (NABP) to increase adherence of antigens containing DNA or RNA. As a source of antigens, we have used supernatants of Jurkat cells undergoing apoptosis in vitro. As results presented show, a poly-L-lysine (PLL) pre-coat significantly enhances detection of antibodies to DNA as well as antigens such as histones, SSA, SSB and RNP. Comparison of the ELISA assay with the PLL pre-coat with a multiplex assay using the BioPlex® 2200 system indicated good agreement in results for a panel of lupus sera. Together, these studies indicate that a pre-coat with a positively charged polymer can increase the sensitivity of an ANA ELISA using as antigens molecules released from dead and dying cells. This assay platform may facilitate ANA testing by providing an ensemble of antigens more similar in composition and structure with antigens present in vivo, with a NABP promoting adherence via charge-charge interactions.

Authors
Stearns, NA; Zhou, S; Petri, M; Binder, SR; Pisetsky, DS
MLA Citation
Stearns, NA, Zhou, S, Petri, M, Binder, SR, and Pisetsky, DS. "The Use of Poly-L-Lysine as a Capture Agent to Enhance the Detection of Antinuclear Antibodies by ELISA." PloS one 11.9 (January 2016): e0161818-.
PMID
27611194
Source
epmc
Published In
PloS one
Volume
11
Issue
9
Publish Date
2016
Start Page
e0161818
DOI
10.1371/journal.pone.0161818

Oligonucleotide-Based Drug Development

Authors
Wang, J; Lon, H-K; Lee, S-L; Burckart, GJ; Pisetsky, DS
MLA Citation
Wang, J, Lon, H-K, Lee, S-L, Burckart, GJ, and Pisetsky, DS. "Oligonucleotide-Based Drug Development." Therapeutic Innovation & Regulatory Science 49.6 (November 2015): 861-868.
Source
crossref
Published In
Therapeutic Innovation and Regulatory Science
Volume
49
Issue
6
Publish Date
2015
Start Page
861
End Page
868
DOI
10.1177/2168479015592195

The Role of Cell Death in the Pathogenesis of SLE: Is Pyroptosis the Missing Link?

Systemic lupus erythematosus (SLE) is a prototypic autoimmune disease characterized by the production of antinuclear antibodies (ANAs) in association with systemic inflammation and organ damage. In addition to genetic factors, a contribution of infection to disease induction has been proposed. In the pathogenesis of lupus, immune complexes of ANAs with nuclear antigens can form and both deposit in the tissue and stimulate cytokine production to intensify inflammation. As such, the extracellular release of nuclear antigens to form pathogenic immune complexes is an important step in the pathway to disease. This release has been considered the consequence of cell death, with apoptotic cells the relevant source of nuclear material. While apoptosis could serve this role, other death forms may act similarly. Among these death forms, pyroptosis, which can be induced by inflammasome activation during infection, has features suggesting involvement in lupus. Thus, unlike apoptosis, pyroptosis is a pro-inflammatory process. Furthermore, pyroptosis leads to the release of intracellular contents including HMGB1 and ATP, both of which can act as DAMPs (death associated molecular patterns) to stimulate further inflammation. Importantly, pyroptosis can lead to the release of intact nuclei, suggesting a relationship to events in the formation of LE cells. While the role of pyroptosis in SLE is hypothetical at this time, further analysis of this death form should provide new insights into lupus pathogenesis and provide the missing link between infection and the initiation of lupus by products of dead and dying cells.

Authors
Magna, M; Pisetsky, DS
MLA Citation
Magna, M, and Pisetsky, DS. "The Role of Cell Death in the Pathogenesis of SLE: Is Pyroptosis the Missing Link?." Scandinavian journal of immunology 82.3 (September 2015): 218-224. (Review)
PMID
26118732
Source
epmc
Published In
Scandinavian Journal of Immunology
Volume
82
Issue
3
Publish Date
2015
Start Page
218
End Page
224
DOI
10.1111/sji.12335

CD40L expression in plasma of volunteers following LPS administration: A comparison between assay of CD40L on platelet microvesicles and soluble CD40L.

CD40 ligand (CD40L) is a transmembrane protein that is mainly expressed on activated T cells and platelets. This protein, however, may also be shed from cells and circulate in the blood in a soluble form. "Soluble CD40L" has attracted interest as a biomarker as it can interact with CD40 and elicit cellular responses involved in the pathophysiology of various thrombotic and inflammatory conditions. As platelets can release microvesicles following activation, we investigated the expression of CD40L on circulating microvesicles as well as CD40L in plasma, in an experimental model of inflammation in healthy volunteers (i.e., intravenous lipopolysaccharide administration). We studied CD40L quantified as CD40L-positive platelet microvesicles by flow cytometry, and as CD40L in plasma ("soluble CD40L") by an ELISA. Results of these studies showed that levels of CD40L exposed on platelet microvesicles were significantly increased after lipopolysaccharide administration. ELISA measurements of CD40L in plasma ("soluble CD40L") did not show any significant increase in plasma levels over time. Separation of soluble and vesicle-bound CD40L by high-speed centrifugation indicated that the ELISA can also detect CD40L on microvesicles, as a trend toward increased concentrations were observed in the pellet of high-speed centrifuged samples (i.e., in samples in which microvesicles are enriched). Together, these findings suggest that platelet microvesicles are a source of CD40L in the circulation and that CD40L exposure on platelet microvesicles increases following experimentally induced inflammation. Our data also suggest that determining levels of CD40L on microvesicles in plasma samples may provide a more sensitive detection of changes in CD40L expression than measurement of "soluble CD40L" in plasma with an ELISA. In addition, information regarding the cellular source of CD40L can be obtained with a flow cytometry-based microvesicle assay in a way not possible with an ordinary ELISA.

Authors
Mobarrez, F; Sjövik, C; Soop, A; Hållström, L; Frostell, C; Pisetsky, DS; Wallén, H
MLA Citation
Mobarrez, F, Sjövik, C, Soop, A, Hållström, L, Frostell, C, Pisetsky, DS, and Wallén, H. "CD40L expression in plasma of volunteers following LPS administration: A comparison between assay of CD40L on platelet microvesicles and soluble CD40L." Platelets 26.5 (January 2015): 486-490.
PMID
24964251
Source
epmc
Published In
Platelets (Informa)
Volume
26
Issue
5
Publish Date
2015
Start Page
486
End Page
490
DOI
10.3109/09537104.2014.932339

Rheumatoid vasculitis: going, going, but not yet gone.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Rheumatoid vasculitis: going, going, but not yet gone." Arthritis research & therapy 17 (January 2015): 116-.
PMID
25952359
Source
epmc
Published In
Arthritis Research and Therapy
Volume
17
Publish Date
2015
Start Page
116
DOI
10.1186/s13075-015-0635-0

Relevant concepts of functioning for patients with systemic lupus erythematosus identified in a Delphi exercise of experts and a literature review.

OBJECTIVE: To identify in a Delphi exercise of international systemic lupus erythematosus (SLE) experts and a systematic literature review the most relevant concepts that impact on the functioning of SLE patients. METHODS: Sixty SLE experts participated in all rounds of a 3-round e-mail-based Delphi exercise; for the literature review, 573 manuscripts out of 4 decades were analyzed. Concepts in the first Delphi round and from the literature were linked to categories of the International Classification of Functioning, Disability and Health (ICF). Categories were voted on individually in a feedback-driven Delphi process, and ranked by frequency in the literature, respectively. RESULTS: In the Delphi exercise, at least 80% of the participants found 30 categories of the domain body functions and structures, and 3 categories in the domain activities and participation and environmental factors important. In general, the categories identified in the literature review overlapped with those in the Delphi exercise with regard to body functions and structures, while showing some differences in other domains. The highest agreement concerned the ICF categories "joints," "skin and related structures," "fatiguability," "immunological system functions," and "handling stress and other psychological demands." Agreement with an earlier patient Delphi exercise was considerable. CONCLUSION: A 3-step Delphi exercise of 60 SLE experts and a literature review identified a wide spectrum of relevant ICF categories with impact on functioning of SLE patients. The categories derived from these approaches overlap with each other and with those of a patient Delphi exercise.

Authors
Leuchten, N; Bauernfeind, B; Kuttner, J; Stamm, T; Smolen, JS; Pisetsky, DS; Aringer, M
MLA Citation
Leuchten, N, Bauernfeind, B, Kuttner, J, Stamm, T, Smolen, JS, Pisetsky, DS, and Aringer, M. "Relevant concepts of functioning for patients with systemic lupus erythematosus identified in a Delphi exercise of experts and a literature review." Arthritis care & research 66.12 (December 2014): 1895-1904. (Review)
PMID
24839085
Source
epmc
Published In
Arthritis Care and Research
Volume
66
Issue
12
Publish Date
2014
Start Page
1895
End Page
1904
DOI
10.1002/acr.22372

The complex role of DNA, histones and HMGB1 in the pathogenesis of SLE.

Systemic lupus erythematosus (SLE) is a prototypic autoimmune disease characterized by the production of antinuclear antibodies (ANA) in association with protean clinic manifestations. ANA can bind to nuclear molecules, most prominently DNA and histones in nucleosomes, to form complexes to promote pathogenesis. Because of the intrinsic immunological activity of the nuclear components, these complexes can amplify responses by interacting with diverse pattern recognition receptors and internal sensing systems. Among molecules associated with nucleosomal components, HMGB1, a non-histone protein, can emanate from activated and dying cells; HMGB1's immune activity is determined by post-translational modifications, redox state, and binding to other immune mediators. Although ANAs form complexes that deposit in the kidney or induce type 1 interferon, ANAs may also block immune activity. Together, these studies highlight the importance of complexes in the pathogenesis of lupus and their role as antigens, immunogens, and adjuvants.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "The complex role of DNA, histones and HMGB1 in the pathogenesis of SLE." Autoimmunity 47.8 (December 2014): 487-493. (Review)
PMID
24916843
Source
epmc
Published In
Autoimmunity (Informa)
Volume
47
Issue
8
Publish Date
2014
Start Page
487
End Page
493
DOI
10.3109/08916934.2014.921811

The role of antigen specificity in the binding of murine monoclonal anti-DNA antibodies to microparticles from apoptotic cells.

Antibodies to DNA (anti-DNA) are the serological hallmark of systemic lupus erythematosus and markers of underlying immune system disturbances. These antibodies bind to both single-stranded and double-stranded DNA, mediating pathogenesis by forming immune complexes. As shown recently, DNA in blood exists in both free and particulate forms, with DNA representing an important component of microparticles. Microparticles are membrane-bound vesicles containing nuclear molecules, released by membrane blebbing during cell death and activation. A panel of monoclonal NZB/NZW F1 anti-DNA antibodies was tested for binding to microparticles generated from apoptotic THP-1 and Jurkat cells. These studies showed that only certain anti-DNA antibodies in the panel, specific for double-stranded DNA, bound to microparticles. Binding to particles was reduced by soluble DNA or DNase treatment. Together, these results indicate that particle binding is a feature of only certain anti-DNA antibodies, reflecting immunochemical properties of the antibodies and the nature of the exposed DNA antigens.

Authors
Ullal, AJ; Marion, TN; Pisetsky, DS
MLA Citation
Ullal, AJ, Marion, TN, and Pisetsky, DS. "The role of antigen specificity in the binding of murine monoclonal anti-DNA antibodies to microparticles from apoptotic cells." Clinical immunology (Orlando, Fla.) 154.2 (October 2014): 178-187.
PMID
24873886
Source
epmc
Published In
Clinical Immunology
Volume
154
Issue
2
Publish Date
2014
Start Page
178
End Page
187
DOI
10.1016/j.clim.2014.05.007

The expression of cytokines and chemokines in the blood of patients with severe weight loss from anorexia nervosa: an exploratory study.

Anorexia nervosa (AN) is a serious, potentially life-threatening disorder characterized by severe weight loss, dysregulated eating, and often excessive exercise. While psychiatric illnesses such as depression are associated with increased levels of pro-inflammatory mediators, evidence for such disturbances in patients with AN has been less clear. In an exploratory study of possible disturbances in immune responses in AN, we assayed a panel of cytokines and chemokines in the blood of patients undergoing inpatient treatment, testing the hypothesis that metabolic disturbances in this disease would lead to a pattern of immune disturbances distinct from that of other psychiatric diseases. For this purpose, we evaluated patients by the Beck Depression Inventory-II (BDI-II) and the Eating Disorders Examination-Questionnaire and assessed cytokines and chemokines by enzyme-linked immunosorbent assays. Patients reported a moderate level of depression (mean BDI-II = 22.6) but exhibited few immunologic abnormalities of the kind associated with major depressive disorder [e.g., increased interleukin (IL)-6]; RANTES showed the most frequent elevations and was increased in 4 of the patients studied. Together, these findings suggest that features of AN such as loss of adipose tissue and excessive exercise may attenuate cytokine production and thus modulate the experience of illness that impacts on core features of disease.

Authors
Pisetsky, DS; Trace, SE; Brownley, KA; Hamer, RM; Zucker, NL; Roux-Lombard, P; Dayer, J-M; Bulik, CM
MLA Citation
Pisetsky, DS, Trace, SE, Brownley, KA, Hamer, RM, Zucker, NL, Roux-Lombard, P, Dayer, J-M, and Bulik, CM. "The expression of cytokines and chemokines in the blood of patients with severe weight loss from anorexia nervosa: an exploratory study." Cytokine 69.1 (September 2014): 110-115.
PMID
25022969
Source
epmc
Published In
Cytokine
Volume
69
Issue
1
Publish Date
2014
Start Page
110
End Page
115
DOI
10.1016/j.cyto.2014.05.018

The expression of HMGB1 on microparticles from Jurkat and HL-60 cells undergoing apoptosis in vitro.

HMGB1 is a highly conserved nuclear protein that displays important biological activities inside as well as outside the cell and serves as a prototypic alarmin to activate innate immunity. The translocation of HMGB1 from inside to outside the cell occurs with cell activation as well as cell death, including apoptosis. Apoptosis is also a setting for the release of cellular microparticles (MPs), which are small membrane-bound vesicles that represent an important source of extracellular nuclear molecules. To investigate whether HMGB1 released from cells during apoptosis is also present on MPs, we determined the presence of HMGB1 on particles released from Jurkat and HL-60 cells induced to undergo apoptosis in vitro by treatment with either etoposide or staurosporine; MPs released from cells undergoing necrosis by freeze-thaw were also characterized. As shown by both Western blot analysis and flow cytometry, MPs from apoptotic cells contain HMGB1, with binding by antibodies indicating an accessible location in the particle structure. These results indicate that HMGB1, like other nuclear molecules, can translocate into MPs during apoptosis and demonstrate another biochemical form of this molecule that may be immunologically active.

Authors
Spencer, DM; Mobarrez, F; Wallén, H; Pisetsky, DS
MLA Citation
Spencer, DM, Mobarrez, F, Wallén, H, and Pisetsky, DS. "The expression of HMGB1 on microparticles from Jurkat and HL-60 cells undergoing apoptosis in vitro." Scandinavian journal of immunology 80.2 (August 2014): 101-110.
PMID
24846056
Source
epmc
Published In
Scandinavian Journal of Immunology
Volume
80
Issue
2
Publish Date
2014
Start Page
101
End Page
110
DOI
10.1111/sji.12191

The effect of polyamines on the binding of anti-DNA antibodies from patients with SLE and normal human subjects.

Antibodies to DNA (anti-DNA) are the serological hallmark of systemic lupus erythematosus (SLE). To elucidate specificity further, the effect of polyamines on the binding of anti-DNA antibodies from patients with lupus was tested by ELISA to calf thymus (CT) DNA; we also assessed the binding of plasmas of patients and normal human subjects (NHS) to Micrococcus luteus (MC) DNA. As these studies showed, spermine can dose-dependently inhibit SLE anti-DNA binding to CT DNA and can promote dissociation of preformed immune complexes. With MC DNA as antigen, spermine failed to inhibit the NHS anti-DNA binding. Studies using plasmas adsorbed to a CT DNA cellulose affinity indicated that SLE plasmas are mixtures of anti-DNA that differ in inhibition by spermine and binding to conserved and non-conserved determinants. Together, these studies demonstrate that spermine can influence the binding of anti-DNA autoantibodies and may contribute to the antigenicity of DNA.

Authors
Wang, X; Stearns, NA; Li, X; Pisetsky, DS
MLA Citation
Wang, X, Stearns, NA, Li, X, and Pisetsky, DS. "The effect of polyamines on the binding of anti-DNA antibodies from patients with SLE and normal human subjects." Clinical immunology (Orlando, Fla.) 153.1 (July 2014): 94-103.
PMID
24732074
Source
epmc
Published In
Clinical Immunology
Volume
153
Issue
1
Publish Date
2014
Start Page
94
End Page
103
DOI
10.1016/j.clim.2014.04.003

The expression of HMGB1 on microparticles released during cell activation and cell death in vitro and in vivo.

High mobility group box protein 1 (HMGB1) is a nonhistone nuclear protein that is a prototypic alarmin that can stimulate innate immunity and drive the pathogenesis of a wide range of inflammatory diseases. While HMGB1 can be released from both activated and dying cells, its biochemical and immunological properties differ depending on the release mechanism, resulting from redox changes and posttranslational modifications including acetylation. In addition to release of HMGB1, cell death is associated with the release of microparticles. Microparticles are small membrane-bound vesicles that contain cytoplasmic, nuclear and membrane components. Like HMGB1, microparticles display immunological activity and levels are elevated in diseases characterized by inflammation and vasculopathy. While studies have addressed the immunological effects of HMGB1 and microparticles independently, HMGB1, like other nuclear molecules, is a component of microparticles. Evidence for the physical association of HMGB1 comes from Western blot analysis of microparticles derived from RAW 264.7 macrophage cells stimulated by lipopolysaccharide (LPS) or induced to undergo apoptosis by treatment with etoposide or staurosporine in vitro. Analysis of microparticles in the blood of healthy volunteers receiving LPS shows the presence of HMGB1 as assessed by flow cytometry. Together, these findings indicate that HMGB1 can be a component of microparticles and may contribute to their activities. Furthermore, particle HMGB1 may represent a useful biomarker for in vivo events that may not be reflected by measurement of the total amount of HMGB1 in the blood.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "The expression of HMGB1 on microparticles released during cell activation and cell death in vitro and in vivo." Molecular Medicine 20 (April 2014): 158-163. (Review)
PMID
24618884
Source
epmc
Published In
Molecular medicine (Cambridge, Mass.)
Volume
20
Publish Date
2014
Start Page
158
End Page
163
DOI
10.2119/molmed.2014.00014

The properties of microparticles from RAW 264.7 macrophage cells undergoing in vitro activation or apoptosis.

Microparticles (MPs) are small, membrane-bound vesicles that arise from dead and dying cells, and display pro-inflammatory and pro-thrombotic activity. As shown previously, the RAW 264.7 murine macrophage cell line can release MPs following stimulation with LPS or polyinosinic:polycytidylic acid [poly (I:C)], ligands of TLR4 and TLR3 respectively. To determine the relationship of these MPs to those released during apoptosis, the nucleic acid content of MPs from cultures stimulated with LPS or poly (I:C) was compared with the nucleic acid content of MPs from untreated cells or cells induced to undergo apoptosis by treatment with etoposide or staurosporine (STS). As results of these studies showed, MPs from activated, apoptotic and untreated cells had features in common, as demonstrated by binding of the nucleic acid dyes SYTO 13 and propidium iodide; molecular mass of DNA; and binding of monoclonal anti-DNA and anti-nucleosome Abs. While MPs from the different culture conditions all contained ribosomal RNA, ribosomal RNA from MPs from STS-treated cells showed cleavage and degradation. Taken together, these studies indicate that the nucleic acid content of MPs from activated and apoptotic cells have important similarities, suggesting that events during TLR activation may lead to apoptosis and subsequent MP release.

Authors
Spencer, DM; Gauley, J; Pisetsky, DS
MLA Citation
Spencer, DM, Gauley, J, and Pisetsky, DS. "The properties of microparticles from RAW 264.7 macrophage cells undergoing in vitro activation or apoptosis." Innate Immun 20.3 (April 2014): 239-248.
PMID
23839527
Source
pubmed
Published In
Innate Immunity
Volume
20
Issue
3
Publish Date
2014
Start Page
239
End Page
248
DOI
10.1177/1753425913492552

The role of HMGB1 in the pathogenesis of inflammatory and autoimmune diseases.

High-mobility group box 1 (HMGB1) protein is a highly abundant protein that can promote the pathogenesis of inflammatory and autoimmune diseases once it is in an extracellular location. This translocation can occur with immune cell activation as well as cell death, with the conditions for release associated with the expression of different isoforms. These isoforms result from post-translational modifications, with the redox states of three cysteines at positions 23, 45 and 106 critical for activity. Depending on the redox states of these residues, HMGB1 can induce cytokine production via toll-like receptor 4 (TLR4) or promote chemotaxis by binding the chemokine CXCL12 for stimulation via CXCR4. Fully oxidized HMGB1 is inactive. During the course of inflammatory disease, HMGB1 can therefore play a dynamic role depending on its redox state. As a mechanism to generate alarmins, cell death is an important source of HMGB1, although each major cell death form (necrosis, apoptosis, pyroptosis and NETosis) can lead to different isoforms of HMGB1 and variable levels of association of HMGB1 with nucleosomes. The association of HMGB1 with nucleosomes may contribute to the pathogenesis of systemic lupus erythematosus by producing nuclear material whose immunological properties are enhanced by the presence of an alarmin. Since HMGB1 levels in blood or tissue are elevated in many inflammatory and autoimmune diseases, this molecule can serve as a unique biomarker as well as represent a target of novel therapies to block its various activities.

Authors
Magna, M; Pisetsky, DS
MLA Citation
Magna, M, and Pisetsky, DS. "The role of HMGB1 in the pathogenesis of inflammatory and autoimmune diseases." Molecular Medicine 20 (March 24, 2014): 138-146. (Review)
PMID
24531836
Source
epmc
Published In
Molecular medicine (Cambridge, Mass.)
Volume
20
Publish Date
2014
Start Page
138
End Page
146
DOI
10.2119/molmed.2013.00164

The translocation of nuclear molecules during inflammation and cell death.

SIGNIFICANCE: Inflammation is a complex biological process that represents the body's response to infection and/or injury. Endogenous molecules that induce inflammation are called death- or damage-associated molecular patterns (DAMPs). Among cellular constituents with DAMP activity, nuclear molecules can stimulate pattern recognition receptors, including toll-like receptors (TLRs). Current research is elucidating the translocation of nuclear molecules during cell death and identifying novel anti-inflammatory approaches to block their DAMP activity. RECENT ADVANCES: High mobility group box protein 1 (HMGB1), a non-histone nuclear protein, can translocate from cells during immune cell activation and cell death. Depending on redox state, HMGB1 can interact with TLR4 although it can bind to molecules such as cytokines to trigger other receptors. DNA and histones, which are bound together in the nucleus, also have important immunological activity. For DNA, DAMP activity may vary depending upon the binding to molecules that affect cell entry and intracellular location. The role of nuclear molecules in disease has been established in animal models using antibodies as inhibitors. CRITICAL ISSUES: Key issues about the DAMP activity of nuclear molecules relate to (i) the impact on function of biochemical modifications such as redox state and post-translational modification, and (ii) the composition and properties of complexes that nuclear molecules may form with other blood components to affect immunological activity. FUTURE DIRECTIONS: With the recognition of the immunological activity of the products of dead cells, future studies will define the diversity and properties of nuclear molecules in the extracellular space and develop strategies to block their activity during inflammation.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "The translocation of nuclear molecules during inflammation and cell death." Antioxid Redox Signal 20.7 (March 1, 2014): 1117-1125. (Review)
PMID
23373769
Source
pubmed
Published In
Antioxidants & Redox Signaling
Volume
20
Issue
7
Publish Date
2014
Start Page
1117
End Page
1125
DOI
10.1089/ars.2012.5143

The effects of smoking on levels of endothelial progenitor cells and microparticles in the blood of healthy volunteers.

BACKGROUND: Cigarette smoking, both active and passive, is one of the leading causes of morbidity and mortality in cardiovascular disease. To assess the impact of brief smoking on the vasculature, we determined levels of circulating endothelial progenitor cells (EPCs) and circulating microparticles (MPs) following the smoking of one cigarette by young, healthy intermittent smokers. MATERIALS AND METHODS: 12 healthy volunteers were randomized to either smoking or not smoking in a crossover fashion. Blood sampling was performed at baseline, 1, 4 and 24 hours following smoking/not smoking. The numbers of EPCs and MPs were determined by flow cytometry. MPs were measured from platelets, leukocytes and endothelial cells. Moreover, MPs were also labelled with anti-HMGB1 and SYTO 13 to assess the content of nuclear molecules. RESULTS: Active smoking of one cigarette caused an immediate and significant increase in the numbers of circulating EPCs and MPs of platelet-, endothelial- and leukocyte origin. Levels of MPs containing nuclear molecules were increased, of which the majority were positive for CD41 and CD45 (platelet- and leukocyte origin). CD144 (VE-cadherin) or HMGB1 release did not significantly change during active smoking. CONCLUSION: Brief active smoking of one cigarette generated an acute release of EPC and MPs, of which the latter contained nuclear matter. Together, these results demonstrate acute effects of cigarette smoke on endothelial, platelet and leukocyte function as well as injury to the vascular wall.

Authors
Mobarrez, F; Antoniewicz, L; Bosson, JA; Kuhl, J; Pisetsky, DS; Lundbäck, M
MLA Citation
Mobarrez, F, Antoniewicz, L, Bosson, JA, Kuhl, J, Pisetsky, DS, and Lundbäck, M. "The effects of smoking on levels of endothelial progenitor cells and microparticles in the blood of healthy volunteers." PLoS ONE 9.2 (January 2014): e90314-.
PMID
24587320
Source
epmc
Published In
PloS one
Volume
9
Issue
2
Publish Date
2014
Start Page
e90314
DOI
10.1371/journal.pone.0090314

Tumor necrosis factor: is it time to change the name?

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Tumor necrosis factor: is it time to change the name?." Arthritis research & therapy 16.2 (January 2014): 108-.
PMID
25166863
Source
epmc
Published In
Arthritis Research and Therapy
Volume
16
Issue
2
Publish Date
2014
Start Page
108
DOI
10.1186/ar4541

Gout, tophi and the wonders of NETs.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Gout, tophi and the wonders of NETs." Arthritis research & therapy 16.5 (January 2014): 431-.
PMID
25606589
Source
epmc
Published In
Arthritis Research and Therapy
Volume
16
Issue
5
Publish Date
2014
Start Page
431
DOI
10.1186/s13075-014-0431-2

Immune activation by histones: plusses and minuses in inflammation.

Histones are highly cationic proteins that are essential components of the cell nucleus, interacting with DNA to form the nucleosome and regulating transcription. Histones, however, can transit from the cell nucleus during cell death and, once in an extracellular location, can serve as danger signals and activate immune cells. An article in this issue of the European Journal of Immunology [Eur. J. Immunol. 2013. 43: 3336-3342] reports that histones can activate monocyte-derived DCs via the NRLP3 inflammasome to induce the production of IL-1β. As such, histones, which can also stimulate TLRs, may drive events in the immunopathogenesis of a wide range of acute and chronic diseases marked by sterile inflammation. While the mechanism of this stimulation is not known, the positive charge of histones may provide a structural element to promote interaction with cells and activation of downstream signaling systems.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Immune activation by histones: plusses and minuses in inflammation." Eur J Immunol 43.12 (December 2013): 3163-3166.
PMID
24165954
Source
pubmed
Published In
European Journal of Immunology
Volume
43
Issue
12
Publish Date
2013
Start Page
3163
End Page
3166
DOI
10.1002/eji.201344175

Assessment of gene expression in peripheral blood using RNAseq before and after weight restoration in anorexia nervosa.

We examined gene expression in the blood of six females with anorexia nervosa (AN) before and after weight restoration using RNAseq. AN cases (aged 19-39) completed clinical assessments and had blood drawn for RNA at hospital admission (T1,<~75% ideal body weight, IBW) and again at discharge (T2,≥ ~ 85% IBW). To examine the relationship between weight restoration and differential gene expression, normalized gene expression levels were analyzed using a paired design. We found 564 genes whose expression was nominally significantly different following weight restoration (p<0.01, 231 increased and 333 decreased). With a more stringent significance threshold (false discovery rate q<0.05), 67 genes met criteria for differential expression. Of the top 20 genes, CYP11A1, C16orf11, LINC00235, and CPA3 were down-regulated more than two-fold after weight restoration while multiple olfactory receptor genes (OR52J3, OR51L1, OR51A4, and OR51A2) were up-regulated more than two-fold after weight restoration. Pathway analysis revealed up-regulation of two broad pathways with largely overlapping genes, one related to protein secretion and signaling and the other associated with defense response to bacterial regulation. Although results are preliminary secondary to a small sample size, these data provide initial evidence of transcriptional alterations during weight restoration in AN.

Authors
Kim, Y; Trace, SE; Crowley, JJ; Brownley, KA; Hamer, RM; Pisetsky, DS; Sullivan, PF; Bulik, CM
MLA Citation
Kim, Y, Trace, SE, Crowley, JJ, Brownley, KA, Hamer, RM, Pisetsky, DS, Sullivan, PF, and Bulik, CM. "Assessment of gene expression in peripheral blood using RNAseq before and after weight restoration in anorexia nervosa." Psychiatry Res 210.1 (November 30, 2013): 287-293.
PMID
23778302
Source
pubmed
Published In
Psychiatry Research
Volume
210
Issue
1
Publish Date
2013
Start Page
287
End Page
293
DOI
10.1016/j.psychres.2013.05.026

The immunopathogenesis and immunopathology of systemic lupus erythematosus

© 2012 Springer Science+Business Media New York. All rights are reserved. Systemic lupus erythematosus (SLE) is a prototypic autoimmune disease characterized by the production of antinuclear antibodies (ANAs). These antibodies can bind protein and nucleic acid components of the cell nucleus to form immune complexes to drive pathogenesis. These complexes can deposit in the kidney to incite nephritis or stimulate plasmacytoid dendritic cells (pDCs) to produce cytokines, most prominently type 1 interferon. The activity of the complexes reflects the intrinsic immunostimulatory properties of nucleic acids, both DNA and RNA, which can activate the innate immune system by interaction with toll-like receptors (TLRs) as well as non-TLR sensors. The operation of this system requires the presence of extracellular nucleic acids which can be derived from apoptotic and necrotic cells. With impaired clearance of dead and dying cells, levels of extracellular nucleic acids can rise and intensify immune disturbances key to autoimmunity.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "The immunopathogenesis and immunopathology of systemic lupus erythematosus." Lupus Erythematosus: Clinical Evaluation and Treatment. September 1, 2013. 13-26.
Source
scopus
Publish Date
2013
Start Page
13
End Page
26
DOI
10.1007/978-1-4614-1189-5_2

Predictors of preterm birth in patients with mild systemic lupus erythematosus.

OBJECTIVE: While increased disease activity is the best predictor of preterm birth in women with systemic lupus erythematosus (SLE), even women with low disease activity are at increased risk of this complication. Biomarkers that would identify at-risk pregnancies could allow interventions to prevent preterm birth. METHOD: Measures of SLE activity, inflammation, placental health and renal function between 20 and 28 weeks gestation (mid-gestation) were correlated to preterm birth and gestational age at delivery in a prospective cohort of pregnant women with SLE. RESULT: Of the 40 pregnancies in 39 women, all with mild-moderate SLE disease, 9 (23.7%) of the 38 live births were delivered preterm. Low C4 was the only marker of SLE activity associated with younger gestational age at delivery. Elevated ferritin and lower oestradiol correlated with younger gestational age at delivery. Renal function remained normal during all pregnancies at mid-gestation and did not correlate with preterm birth. Higher serum uric acid, however, correlated with younger gestational age at delivery. CONCLUSIONS: In women with SLE with mild-moderate disease activity, ferritin, oestradiol and uric acid levels at mid-gestation may predict preterm birth. These markers may prove to be clinically useful in identifying pregnancies at particularly high risk for adverse outcomes.

Authors
Clowse, MEB; Wallace, DJ; Weisman, M; James, A; Criscione-Schreiber, LG; Pisetsky, DS
MLA Citation
Clowse, MEB, Wallace, DJ, Weisman, M, James, A, Criscione-Schreiber, LG, and Pisetsky, DS. "Predictors of preterm birth in patients with mild systemic lupus erythematosus." Ann Rheum Dis 72.9 (September 1, 2013): 1536-1539.
PMID
23361085
Source
pubmed
Published In
Annals of the rheumatic diseases
Volume
72
Issue
9
Publish Date
2013
Start Page
1536
End Page
1539
DOI
10.1136/annrheumdis-2012-202449

The danger of sex and death in Scarf1(-/-) autoimmune mice.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "The danger of sex and death in Scarf1(-/-) autoimmune mice." Nat Immunol 14.9 (September 2013): 888-889.
PMID
23959178
Source
pubmed
Published In
Nature Immunology
Volume
14
Issue
9
Publish Date
2013
Start Page
888
End Page
889
DOI
10.1038/ni.2684

The Choosing Wisely initiative: does it have your back?

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "The Choosing Wisely initiative: does it have your back? (Published online)." Arthritis Res Ther 15.4 (August 22, 2013): 117-.
PMID
23998494
Source
pubmed
Published In
Arthritis Research and Therapy
Volume
15
Issue
4
Publish Date
2013
Start Page
117
DOI
10.1186/ar4257

Modeling nuclear molecule release during in vitro cell death.

The extracellular release of nuclear molecules occurs prominently during cell death and can determine the immune properties of dead and dying cells. Depending on inciting events and environmental conditions, cells can die by apoptosis or necrosis, although these processes differ in their immunological consequences. Whereas apoptosis is immunologically silent, necrosis leads to inflammation, possibly reflecting the array of "danger" molecules released. To investigate these processes, the extracellular release of HMGB1 during necrosis was characterized in vitro using Jurkat T cell leukemia cells treated to induce necrosis by freeze-thaw, heat, hydrogen peroxide or ethanol. HMGB1 is a non-histone nuclear protein that represents a prototype alarmin; the pro-inflammatory activity of HMGB1, however, depends on past-translational modifications and association with other molecules including cytokines. As results of these studies showed, the extent and kinetics of HMGB1 release from necrotic cells varies markedly depending on its induction. Among treatments tested, freeze-thaw produced the highest levels of extracellular HMGB1; levels with ethanol treatment were very low and in the range of untreated controls. Similar results were obtained with DNA, with freeze-thaw leading to significant amounts of extracellular DNA although this DNA was subject to rapid nuclease digestion. Together, these findings suggest that the rapid release of nuclear molecules is not an invariable feature of necrosis and that the characterization of the immune properties of dead and dying cells should use systems in which the content and stability of extracellular components are well defined.

Authors
Beyer, C; Pisetsky, DS
MLA Citation
Beyer, C, and Pisetsky, DS. "Modeling nuclear molecule release during in vitro cell death." Autoimmunity 46.5 (August 2013): 298-301. (Review)
PMID
23244202
Source
pubmed
Published In
Autoimmunity (Informa)
Volume
46
Issue
5
Publish Date
2013
Start Page
298
End Page
301
DOI
10.3109/08916934.2012.750297

Effects of cartilage impact with and without fracture on chondrocyte viability and the release of inflammatory markers.

Post-traumatic arthritis (PTA) frequently develops after intra-articular fracture of weight bearing joints. Loss of cartilage viability and post-injury inflammation have both been implicated as possible contributing factors to PTA progression. To further investigate chondrocyte response to impact and fracture, we developed a blunt impact model applying 70%, 80%, or 90% surface-to-surface compressive strain with or without induction of an articular fracture in a cartilage explant model. Following mechanical loading, chondrocyte viability, and apoptosis were assessed. Culture media were evaluated for the release of double-stranded DNA (dsDNA) and immunostimulatory activity via nuclear factor kappa B (NF-κB) activity in Toll-like receptor (TLR) -expressing Ramos-Blue reporter cells. High compressive strains, with or without articular fracture, resulted in significantly reduced chondrocyte viability. Blunt impact at 70% strain induced a loss in viability over time through a combination of apoptosis and necrosis, whereas blunt impact above 80% strain caused predominantly necrosis. In the fracture model, a high level of primarily necrotic chondrocyte death occurred along the fracture edges. At sites away from the fracture, viability was not significantly different than controls. Interestingly, both dsDNA release and NF-κB activity in Ramos-Blue cells increased with blunt impact, but was only significantly increased in the media from fractured cores. This study indicates that the mechanism of trauma determines the type of chondrocyte death and the potential for post-injury inflammation.

Authors
Stolberg-Stolberg, JA; Furman, BD; Garrigues, NW; Lee, J; Pisetsky, DS; Stearns, NA; DeFrate, LE; Guilak, F; Olson, SA
MLA Citation
Stolberg-Stolberg, JA, Furman, BD, Garrigues, NW, Lee, J, Pisetsky, DS, Stearns, NA, DeFrate, LE, Guilak, F, and Olson, SA. "Effects of cartilage impact with and without fracture on chondrocyte viability and the release of inflammatory markers." J Orthop Res 31.8 (August 2013): 1283-1292.
PMID
23620164
Source
pubmed
Published In
Journal of Orthopaedic Research
Volume
31
Issue
8
Publish Date
2013
Start Page
1283
End Page
1292
DOI
10.1002/jor.22348

Effect of lipopolysaccharide administration on the number, phenotype and content of nuclear molecules in blood microparticles of normal human subjects.

Microparticles (MPs) are small membrane-bound vesicles that arise from activated and dying cells and promote inflammation and thrombosis. To characterize the in vivo release of MPs, we used flow cytometry to measure MPs in the blood of 15 healthy volunteers administered bacterial endotoxin (lipopolysaccharide or LPS) in the presence of a low dose of hydrocortisone with or without inhaled nitric oxide. MPs, defined as particles less than 1.0 μm in size, were assessed following labelling for CD42a, CD14 and CD62E or CD144 antibodies to identify MPs from platelets (PMPs), monocytes (MMPs) and endothelial cells (EMPs). In addition, PMPs and MMPs were labelled with anti-HMGB1 and stained with SYTO13 to assess nuclear acid content. Administration of LPS led to an increase in the numbers of PMPs, MMPs and EMPs as defined by CD62E, as well as the number of MMPs and PMPs staining with anti-HMGB1 and SYTO13. Inhalation of NO did not influence these findings. Together, these studies show that LPS can increase levels of blood MPs and influence phenotype, including nuclear content. As such, particles may be a source of HMGB1 and other nuclear molecules in the blood during inflammation.

Authors
Soop, A; Hållström, L; Frostell, C; Wallén, H; Mobarrez, F; Pisetsky, DS
MLA Citation
Soop, A, Hållström, L, Frostell, C, Wallén, H, Mobarrez, F, and Pisetsky, DS. "Effect of lipopolysaccharide administration on the number, phenotype and content of nuclear molecules in blood microparticles of normal human subjects." Scand J Immunol 78.2 (August 2013): 205-213.
PMID
23679665
Source
pubmed
Published In
Scandinavian Journal of Immunology
Volume
78
Issue
2
Publish Date
2013
Start Page
205
End Page
213
DOI
10.1111/sji.12076

The role of microparticles in the pathogenesis of rheumatoid arthritis and systemic lupus erythematosus.

Microparticles (MPs) are small membrane-bound vesicles with potent biological activities that can promote the pathogenesis of rheumatoid arthritis and systemic lupus erythematosus (SLE). These particles contain diverse cellular components and are shed from cells during apoptosis or activation. MPs can drive inflammation and autoimmunity by multiple mechanisms reflecting their content of bioactive molecules and ability to engage multiple receptor systems simultaneously. In the rheumatoid joint, particles can stimulate synovitis via their display of cytokines, chemokines, complement and angiogenesis factors. In SLE, particles can serve as an important source of extracellular nuclear molecules to signal 'danger' and form pathogenic immune complexes. Future studies will define the pathways by which particles promote pathogenesis, strategies to block their activity and their utility as biomarkers to assess disease activity and the response to therapy.

Authors
Dye, JR; Ullal, AJ; Pisetsky, DS
MLA Citation
Dye, JR, Ullal, AJ, and Pisetsky, DS. "The role of microparticles in the pathogenesis of rheumatoid arthritis and systemic lupus erythematosus." Scand J Immunol 78.2 (August 2013): 140-148. (Review)
PMID
23672591
Source
pubmed
Published In
Scandinavian Journal of Immunology
Volume
78
Issue
2
Publish Date
2013
Start Page
140
End Page
148
DOI
10.1111/sji.12068

Standardization of anti-DNA antibody assays.

Antibodies to DNA (anti-DNA) are the serological hallmark of systemic lupus erythematosus and represent important biomarkers for clinical and research purposes. These antibodies are part of a family of antibodies to nucleosomes and bind to conserved sites widely present on DNA. While the value of anti-DNA as a biomarker is well established, the assay for these antibodies has involved a variety of DNA sources and systems to detect DNA-anti-DNA interactions. The influence of these variations on antibody detection has complicated assay standardization. As an antigen, DNA has unique features since it is a highly charged polymer that has structural heterogeneity. This heterogeneity can affect antigenicity which can vary on the basis of DNA origin, size, conformation and mobility. In addition, as a polymer, DNA can promote patterns of antibody binding based on monogamous or bivalent interaction which require an extended polynucleotide structure. Understanding the nature of DNA as an antigen can facilitate interpretation of serological tests and underpin efforts at better standardization.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Standardization of anti-DNA antibody assays." Immunol Res 56.2-3 (July 2013): 420-424. (Review)
PMID
23579774
Source
pubmed
Published In
Immunologic Research
Volume
56
Issue
2-3
Publish Date
2013
Start Page
420
End Page
424
DOI
10.1007/s12026-013-8415-x

Translation, treatises, and tweets.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Translation, treatises, and tweets." Arthritis Care Res (Hoboken) 65.6 (June 2013): 839-842.
PMID
23401416
Source
pubmed
Published In
Arthritis Care and Research
Volume
65
Issue
6
Publish Date
2013
Start Page
839
End Page
842
DOI
10.1002/acr.21974

The butterfly rash of lupus: an example of aposematism?

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "The butterfly rash of lupus: an example of aposematism? (Published online)." Arthritis Res Ther 15.1 (February 19, 2013): 106-.
PMID
23425372
Source
pubmed
Published In
Arthritis Research and Therapy
Volume
15
Issue
1
Publish Date
2013
Start Page
106
DOI
10.1186/ar4155

The role of microparticles in the generation of immune complexes in murine lupus.

Systemic lupus erythematosus is a systemic inflammatory disease characterized by antibodies to nuclear molecules in association with immune complex deposition. As shown previously, microparticles (MPs), which are small membrane-bound vesicles released from dying and activated cells, contain nucleic acids and can form immune complexes found in patient blood. To assess the role of MPs in murine lupus, we used flow cytometry to measure the presence of MPs with bound IgG in the blood of MRL-lpr/lpr and NZB/W mice. These studies showed much higher numbers of MPs with bound IgG in the blood of MRL lpr/lpr compared to NZB/W mice. Furthermore, these studies showed that antibodies from MRL-lpr/lpr mice bound better to MPs from apoptotic cells than those from NZB/W mice. Together, these studies indicate important differences in the serological features of the two strains as reflected by the capacity of antibodies to bind to MPs.

Authors
Ullal, AJ; Pisetsky, DS
MLA Citation
Ullal, AJ, and Pisetsky, DS. "The role of microparticles in the generation of immune complexes in murine lupus." Clin Immunol 146.1 (January 2013): 1-9.
PMID
23159786
Source
pubmed
Published In
Clinical Immunology
Volume
146
Issue
1
Publish Date
2013
Start Page
1
End Page
9
DOI
10.1016/j.clim.2012.10.004

Assessment of gene expression in peripheral blood using RNAseq before and after weight restoration in anorexia nervosa

We examined gene expression in the blood of six females with anorexia nervosa (AN) before and after weight restoration using RNAseq. AN cases (aged 19-39) completed clinical assessments and had blood drawn for RNA at hospital admission (T1,<~75% ideal body weight, IBW) and again at discharge (T2,≥ ~ 85% IBW). To examine the relationship between weight restoration and differential gene expression, normalized gene expression levels were analyzed using a paired design. We found 564 genes whose expression was nominally significantly different following weight restoration (p<0.01, 231 increased and 333 decreased). With a more stringent significance threshold (false discovery rate q<0.05), 67 genes met criteria for differential expression. Of the top 20 genes, CYP11A1, C16orf11, LINC00235, and CPA3 were down-regulated more than two-fold after weight restoration while multiple olfactory receptor genes (OR52J3, OR51L1, OR51A4, and OR51A2) were up-regulated more than two-fold after weight restoration. Pathway analysis revealed up-regulation of two broad pathways with largely overlapping genes, one related to protein secretion and signaling and the other associated with defense response to bacterial regulation. Although results are preliminary secondary to a small sample size, these data provide initial evidence of transcriptional alterations during weight restoration in AN. © 2013 Elsevier Ireland Ltd.

Authors
Kim, Y; Trace, SE; Crowley, JJ; Brownley, KA; Hamer, RM; Pisetsky, DS; Sullivan, PF; Bulik, CM
MLA Citation
Kim, Y, Trace, SE, Crowley, JJ, Brownley, KA, Hamer, RM, Pisetsky, DS, Sullivan, PF, and Bulik, CM. "Assessment of gene expression in peripheral blood using RNAseq before and after weight restoration in anorexia nervosa." Psychiatry Research 210.1 (2013): 287-293.
Source
scival
Published In
Psychiatry Research
Volume
210
Issue
1
Publish Date
2013
Start Page
287
End Page
293
DOI
10.1016/j.psychres.2013.05.026

The 2013 ACR meeting: mad macs at the Marina.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "The 2013 ACR meeting: mad macs at the Marina." Arthritis Res Ther 15.6 (2013): 130-.
PMID
24364924
Source
pubmed
Published In
Arthritis Research and Therapy
Volume
15
Issue
6
Publish Date
2013
Start Page
130
DOI
10.1186/ar4419

Standardization of anti-DNA antibody assays

Antibodies to DNA (anti-DNA) are the serological hallmark of systemic lupus erythematosus and represent important biomarkers for clinical and research purposes. These antibodies are part of a family of antibodies to nucleosomes and bind to conserved sites widely present on DNA. While the value of anti-DNA as a biomarker is well established, the assay for these antibodies has involved a variety of DNA sources and systems to detect DNA-anti-DNA interactions. The influence of these variations on antibody detection has complicated assay standardization. As an antigen, DNA has unique features since it is a highly charged polymer that has structural heterogeneity. This heterogeneity can affect antigenicity which can vary on the basis of DNA origin, size, conformation and mobility. In addition, as a polymer, DNA can promote patterns of antibody binding based on monogamous or bivalent interaction which require an extended polynucleotide structure. Understanding the nature of DNA as an antigen can facilitate interpretation of serological tests and underpin efforts at better standardization. © 2013 Springer Science+Business Media New York (outside the USA).

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Standardization of anti-DNA antibody assays." Immunologic Research 56.2-3 (2013): 420-424.
Source
scival
Published In
Immunologic Research
Volume
56
Issue
2-3
Publish Date
2013
Start Page
420
End Page
424
DOI
10.1007/s12026-013-8415-x

Predictors of preterm birth in patients with mild systemic lupus erythematosus

Objective: While increased disease activity is the best predictor of preterm birth in women with systemic lupus erythematosus (SLE), even women with low disease activity are at increased risk of this complication. Biomarkers that would identify at-risk pregnancies could allow interventions to prevent preterm birth. Method Measures of SLE activity, inflammation, placental health and renal function between 20 and 28 weeks gestation (mid-gestation) were correlated to preterm birth and gestational age at delivery in a prospective cohort of pregnant women with SLE. Result Of the 40 pregnancies in 39 women, all with mild-moderate SLE disease, 9 (23.7%) of the 38 live births were delivered preterm. Low C4 was the only marker of SLE activity associated with younger gestational age at delivery. Elevated ferritin and lower oestradiol correlated with younger gestational age at delivery. Renal function remained normal during all pregnancies at mid-gestation and did not correlate with preterm birth. Higher serum uric acid, however, correlated with younger gestational age at delivery. Conclusions: In women with SLE with mild-moderate disease activity, ferritin, oestradiol and uric acid levels at mid-gestation may predict preterm birth. These markers may prove to be clinically useful in identifying pregnancies at particularly high risk for adverse outcomes.

Authors
Clowse, MEB; Wallace, DJ; Weisman, M; James, A; Criscione-Schreiber, LG; Pisetsky, DS
MLA Citation
Clowse, MEB, Wallace, DJ, Weisman, M, James, A, Criscione-Schreiber, LG, and Pisetsky, DS. "Predictors of preterm birth in patients with mild systemic lupus erythematosus." Annals of the Rheumatic Diseases 72.9 (2013): 1536-1539.
Source
scival
Published In
Annals of the rheumatic diseases
Volume
72
Issue
9
Publish Date
2013
Start Page
1536
End Page
1539
DOI
10.1136/annrheumdis-2012-202449

The effectors of innate immunity: DAMPs, DAMEs, or DIMEs?

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "The effectors of innate immunity: DAMPs, DAMEs, or DIMEs?." Arthritis Res Ther 15.5 (2013): 123-.
PMID
24172019
Source
pubmed
Published In
Arthritis Research and Therapy
Volume
15
Issue
5
Publish Date
2013
Start Page
123
DOI
10.1186/ar4363

Microparticles as mediators and biomarkers of rheumatic disease.

Microparticles (MPs) are small membrane-bound vesicles that arise from activated and dying cells and enter the blood to display pro-inflammatory and pro-thrombotic activities. MPs are 0.1-1.0 μm in size and incorporate nuclear, cytoplasmic and membrane molecules as they detach from cells. This process can occur with cell activation as well as cell death, with particles likely corresponding to blebs that form on the cell surface during apoptosis. To measure particle expression, flow cytometry allows determination of particle numbers based on size as well as surface markers that denote the cell of origin; platelet MPs are usually the most abundant type in blood. As shown in in vitro and in vivo systems, MPs can promote inflammation and thrombosis resulting from their content of cytokines like IL-1 and pro-coagulant molecules like tissue factor. Certain particle types can be anti-inflammatory, however, suggesting a range of immunomodulatory activities depending on the cell of origin. Studies on patients with a wide range of rheumatic disease show increased MP numbers in blood, with platelet and endothelial particles associated with vascular manifestations; increased numbers of particles also occur in the joint fluid where they may drive cytokine production and activate synoviocytes. In autoimmune diseases such as SLE and RA, MPs may also contribute to disease pathogenesis by the formation of immune complexes. MPs thus represent novel subcellular structures that can impact on the pathogenesis of rheumatic disease and serve as biomarkers of underlying cellular disturbances.

Authors
Pisetsky, DS; Ullal, AJ; Gauley, J; Ning, TC
MLA Citation
Pisetsky, DS, Ullal, AJ, Gauley, J, and Ning, TC. "Microparticles as mediators and biomarkers of rheumatic disease." Rheumatology (Oxford) 51.10 (October 2012): 1737-1746. (Review)
PMID
22403183
Source
pubmed
Published In
Rheumatology
Volume
51
Issue
10
Publish Date
2012
Start Page
1737
End Page
1746
DOI
10.1093/rheumatology/kes028

The extracellular release of DNA and HMGB1 from Jurkat T cells during in vitro necrotic cell death.

In innate immunity, dead and dying cells release internal constituents that can serve as damage-associated molecular patterns (DAMPs) or alarmins. This release occurs more abundantly during necrosis than apoptosis and may account for the differences in the immunologic properties of these death forms. To elucidate DAMP release in necrosis, we compared the levels of two nuclear molecules (DNA and HMGB1, a non-histone protein with alarmin activity) in media following necrosis of Jurkat T cells by freeze-thawing, ethanol, heat or hydrogen peroxide treatment. In our experiments, DNA release was measured by fluorimetry with the dye PicoGreen, while HMGB1 was measured by Western blotting. As the results of our study show, each form of necrosis is associated with a distinct pattern of DNA and HMGB1 release with respect to kinetics and amounts. Of these, freeze-thawing produced the highest and most rapid increase in HMGB1 and DNA levels, although the released DNA was subject to nuclease digestion; in addition, freeze-thawing led to the production of particles measured by flow cytometry. Together, these results indicate that experimental necrosis leads to diverse patterns of nuclear molecule release which could affect their immunologic activity.

Authors
Beyer, C; Stearns, NA; Giessl, A; Distler, JHW; Schett, G; Pisetsky, DS
MLA Citation
Beyer, C, Stearns, NA, Giessl, A, Distler, JHW, Schett, G, and Pisetsky, DS. "The extracellular release of DNA and HMGB1 from Jurkat T cells during in vitro necrotic cell death." Innate Immun 18.5 (October 2012): 727-737.
PMID
22344226
Source
pubmed
Published In
Innate Immunity
Volume
18
Issue
5
Publish Date
2012
Start Page
727
End Page
737
DOI
10.1177/1753425912437981

4th International Forum on Rheumatoid Arthritis: Great times near the Great Wall.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "4th International Forum on Rheumatoid Arthritis: Great times near the Great Wall. (Published online)" Arthritis Res Ther 14.5 (September 27, 2012): 125-.
PMID
23025638
Source
pubmed
Published In
Arthritis Research and Therapy
Volume
14
Issue
5
Publish Date
2012
Start Page
125
DOI
10.1186/ar4036

Antinuclear antibodies in rheumatic disease: a proposal for a function-based classification.

Antinuclear antibodies (ANAs) are a diverse group of autoantibodies that bind macromolecular components of the cell nucleus. While some ANAs occur in normal individuals, others are expressed almost exclusively in patients with rheumatic disease and serve as markers for diagnosis and prognosis. Despite the clinical associations of ANAs, the relationship of these antibodies to specific disease manifestations is often unknown because the target antigens are intracellular molecules that are ubiquitously expressed. In systemic lupus erythematosus, the role of ANAs in disease manifestations is better understood, especially for antibodies to DNA and related nucleosomal antigens. These antibodies can promote nephritis by the formation of immune complexes that are deposited in the kidney. In addition, anti-DNA, along with antibodies to RNA-binding proteins such as anti-Sm, can induce non-specific immune abnormalities based on the induction of type interferon 1 by plasmacytoid dendritic cells. Despite ANA expression in rheumatic disease, studies in animal models of inflammation and tissue injury indicate that antibodies to certain nuclear molecules such as HMGB1 have protective effects. Together, these considerations suggest a function-based classification of ANAs based on their expression in normal and autoimmune individuals as well as their capacity to induce or attenuate immunological disturbances. This classification provides a framework to elucidate the serological features of rheumatic disease and the often uncertain relationship between ANA expression and disease manifestations.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Antinuclear antibodies in rheumatic disease: a proposal for a function-based classification." Scand J Immunol 76.3 (September 2012): 223-228. (Review)
PMID
22670594
Source
pubmed
Published In
Scandinavian Journal of Immunology
Volume
76
Issue
3
Publish Date
2012
Start Page
223
End Page
228
DOI
10.1111/j.1365-3083.2012.02728.x

The effects of heparins on the liver: application of mechanistic serum biomarkers in a randomized study in healthy volunteers.

Heparins have been reported to cause elevations in serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) but have not been associated with clinically significant liver injury. The mechanisms underlying these benign laboratory abnormalities are unknown. Forty-eight healthy men were randomized to receive subcutaneous injections of unfractionated heparin (UFH; 150 U/kg), enoxaparin sodium (1 mg/kg), dalteparin sodium (120 IU/kg), or adomiparin sodium (125 IU/kg; a novel heparin) every 12 h for 4.5 days. Asymptomatic elevations in serum ALT or AST were observed in >90% of the subjects. Elevations were also observed in the levels of serum sorbitol dehydrogenase (SDH), glutamate dehydrogenase (GLDH), miR-122, high-mobility group box-1 protein (including the acetylated form), full-length keratin 18, and DNA. Keratin 18 fragments, which are apoptosis biomarkers, were not detected. Biomarker profiles did not differ significantly across heparin treatments. We conclude that heparins as a class cause self-limited and mild hepatocyte necrosis with secondary activation of an innate immune response.

Authors
Harrill, AH; Roach, J; Fier, I; Eaddy, JS; Kurtz, CL; Antoine, DJ; Spencer, DM; Kishimoto, TK; Pisetsky, DS; Park, BK; Watkins, PB
MLA Citation
Harrill, AH, Roach, J, Fier, I, Eaddy, JS, Kurtz, CL, Antoine, DJ, Spencer, DM, Kishimoto, TK, Pisetsky, DS, Park, BK, and Watkins, PB. "The effects of heparins on the liver: application of mechanistic serum biomarkers in a randomized study in healthy volunteers." Clin Pharmacol Ther 92.2 (August 2012): 214-220.
PMID
22739141
Source
pubmed
Published In
Clinical Pharmacology & Therapeutics (Nature)
Volume
92
Issue
2
Publish Date
2012
Start Page
214
End Page
220
DOI
10.1038/clpt.2012.40

The origin and properties of extracellular DNA: from PAMP to DAMP.

DNA is a polymeric macromolecule whose biological activities depend on location as well as binding to associated molecules. Inside the cell, DNA is the source of genetic information and binds histones to form nucleosomes. DNA can exit the cell, however, to enter the extracellular space primarily during cell death, either apoptosis or necrosis, as well as NETosis. While bacterial DNA is a potent immune stimulant by virtue of its CpG motifs, mammalian DNA, which is ordinarily inactive, can acquire activity by associating with nuclear, cytoplasmic and serum proteins which promote its uptake into cells to stimulate internal DNA sensors, including Toll-like receptor 9. Among these proteins, anti-DNA autoantibodies can form immune complexes with DNA to stimulate plasmacytoid dendritic cells to produce type 1 interferon. Together, these findings suggest that the immune properties of DNA are mutable and diverse, reflecting its context and the array of attached molecules.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "The origin and properties of extracellular DNA: from PAMP to DAMP." Clin Immunol 144.1 (July 2012): 32-40. (Review)
PMID
22659033
Source
pubmed
Published In
Clinical Immunology
Volume
144
Issue
1
Publish Date
2012
Start Page
32
End Page
40
DOI
10.1016/j.clim.2012.04.006

The LE cell: crime scene or crime stopper?

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "The LE cell: crime scene or crime stopper? (Published online)." Arthritis Res Ther 14.3 (June 26, 2012): 120-.
PMID
22738266
Source
pubmed
Published In
Arthritis Research and Therapy
Volume
14
Issue
3
Publish Date
2012
Start Page
120
DOI
10.1186/ar3878

The value in musculoskeletal care: summary and recommendations.

Authors
Gnatz, SM; Pisetsky, DS; Andersson, GBJ
MLA Citation
Gnatz, SM, Pisetsky, DS, and Andersson, GBJ. "The value in musculoskeletal care: summary and recommendations." PM R 4.5 (May 2012): 379-382.
PMID
22613364
Source
pubmed
Published In
PM&R
Volume
4
Issue
5
Publish Date
2012
Start Page
379
End Page
382
DOI
10.1016/j.pmrj.2012.04.004

Microparticles as autoantigens: making immune complexes big.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Microparticles as autoantigens: making immune complexes big." Arthritis Rheum 64.4 (April 2012): 958-961.
PMID
22237935
Source
pubmed
Published In
Arthritis and Rheumatism
Volume
64
Issue
4
Publish Date
2012
Start Page
958
End Page
961
DOI
10.1002/art.34377

Advances in the treatment of inflammatory arthritis.

The inflammatory arthritides are a diverse group of conditions characterised by joint inflammation which can lead to pain, deformity and disability. Of these diseases, rheumatoid arthritis (RA) and spondyloarthritis are two of the most common. While the clinical and demographic features of these diseases differ, the central role of inflammation in their pathogenesis has allowed the development of highly effective treatment strategies with wide applicability. These strategies include the use of biological agents which target the cytokine tumour necrosis factor (TNF), a key mediator of inflammation. With the advent of effective agents, therapy has become more aggressive, reducing disease activity and allowing, at least in RA, remission in many patients. While the array of available effective treatments is extensive, the use of objective measures of disease activity can guide treatment decisions (treat to target) and lead to improved outcomes.

Authors
Pisetsky, DS; Ward, MM
MLA Citation
Pisetsky, DS, and Ward, MM. "Advances in the treatment of inflammatory arthritis." Best Pract Res Clin Rheumatol 26.2 (April 2012): 251-261. (Review)
PMID
22794097
Source
pubmed
Published In
Best Practice and Research: Clinical Rheumatology
Volume
26
Issue
2
Publish Date
2012
Start Page
251
End Page
261
DOI
10.1016/j.berh.2012.03.001

The value in musculoskeletal care: summary and recommendations.

Authors
Gnatz, SM; Pisetsky, DS; Andersson, GBJ
MLA Citation
Gnatz, SM, Pisetsky, DS, and Andersson, GBJ. "The value in musculoskeletal care: summary and recommendations." Semin Arthritis Rheum 41.5 (April 2012): 741-744.
PMID
22483656
Source
pubmed
Published In
Seminars in Arthritis and Rheumatism
Volume
41
Issue
5
Publish Date
2012
Start Page
741
End Page
744
DOI
10.1016/j.semarthrit.2012.02.007

HMGB1: a smoking gun in lupus nephritis?

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "HMGB1: a smoking gun in lupus nephritis? (Published online)." Arthritis Res Ther 14.2 (March 14, 2012): 112-.
PMID
22423653
Source
pubmed
Published In
Arthritis Research and Therapy
Volume
14
Issue
2
Publish Date
2012
Start Page
112
DOI
10.1186/ar3754

The BRIC-GARN Meeting 2011: of mice and men.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "The BRIC-GARN Meeting 2011: of mice and men. (Published online)" Arthritis Res Ther 14.1 (February 24, 2012): 107-.
PMID
22404910
Source
pubmed
Published In
Arthritis Research and Therapy
Volume
14
Issue
1
Publish Date
2012
Start Page
107
DOI
10.1186/ar3698

Gene expression profiles of RAW264.7 macrophages stimulated with preparations of LPS differing in isolation and purity.

Lipopolysaccharide is a major component of the cell wall of Gram-negative bacteria and a potent stimulator of innate immune response via TLR4. Studies on the LPS action both in vivo and in vitro have used different preparations of LPS, including ultra-pure LPS (LIST) and a less pure but less expensive form (Sigma) isolated from Escherichia coli serotype O111:B4. The difference between the effects of these compounds has not been well studied although this information is important in understanding TLR stimulation. In this study, we compared response of RAW264.7 macrophage cells treated LIST or Sigma LPS for 6 h and 24 h. Gene expression data were analyzed to identify specific genes and pathways that are in common and unique to the two LPS preparations. Seven hundred fifty-five genes were differentially expressed at 6 h in response to Sigma LPS and 973 were differentially expressed following LIST LPS treatment, with 503 in common. At 24 h, Sigma LPS induced or repressed 901 genes while 1646 genes were differentially regulated by LIST LPS treatment; 701 genes were shared by two forms of LPS. Although considerably more genes were differentially expressed in response to LIST LPS, similar molecular pathways and transcriptional networks were activated by the two LPS preparations. We also treated bone marrow-derived macrophages (BMMs) from three strains of mice with different concentrations of LIST and Sigma LPS and showed that BMMs produced more IL-6 and TNF-α in response to LIST LPS at low LPS concentrations but, at higher LPS concentrations, more cytokines were produced in response to stimulation by Sigma LPS. Together, these findings suggest that, despite activation of similar molecular pathways by LIST and Sigma LPS preparations, residual protein impurities in the Sigma LPS preparation may nevertheless influence the transcriptional profile attributed to TLR4 stimulation.

Authors
Rutledge, HR; Jiang, W; Yang, J; Warg, LA; Schwartz, DA; Pisetsky, DS; Yang, IV
MLA Citation
Rutledge, HR, Jiang, W, Yang, J, Warg, LA, Schwartz, DA, Pisetsky, DS, and Yang, IV. "Gene expression profiles of RAW264.7 macrophages stimulated with preparations of LPS differing in isolation and purity." Innate Immun 18.1 (February 2012): 80-88.
PMID
21239457
Source
pubmed
Published In
Innate Immunity
Volume
18
Issue
1
Publish Date
2012
Start Page
80
End Page
88
DOI
10.1177/1753425910393540

HMGB1: a multifunctional alarmin driving autoimmune and inflammatory disease.

HMGB1 is a non-histone nuclear protein that can serve as an alarmin to drive the pathogenesis of inflammatory and autoimmune disease. Although primarily located in the cell nucleus, HMGB1 can translocate to the cytoplasm, as well as the extracellular space, during cell activation and cell death; during activation, HMGB1 can undergo post-translational modifications. The activity of HMGB1 varies with the redox states of the cysteine residues, which are required for binding to TLR4. In addition to stimulating cells directly, HMGB1 can form immunostimulatory complexes with cytokines and other endogenous and exogenous factors. In the synovia of patients with rheumatoid arthritis, as well as animal models of this disease, extranuclear expression of HMGB1 is increased and blockade of HMGB1 expression attenuates disease in animal models. In systemic lupus erythematosus, HMGB1 can be a component of immune complexes containing anti-DNA because of its interaction with DNA. In myositis, expression of HMGB1 is enhanced in inflamed muscle and can perturb muscle function. Together, these findings indicate that HMGB1 might be an important mediator and biomarker in rheumatic diseases as well as a target of new therapy.

Authors
Harris, HE; Andersson, U; Pisetsky, DS
MLA Citation
Harris, HE, Andersson, U, and Pisetsky, DS. "HMGB1: a multifunctional alarmin driving autoimmune and inflammatory disease. (Published online)" Nat Rev Rheumatol 8.4 (January 31, 2012): 195-202. (Review)
PMID
22293756
Source
pubmed
Published In
Nature Reviews Rheumatology
Volume
8
Issue
4
Publish Date
2012
Start Page
195
End Page
202
DOI
10.1038/nrrheum.2011.222

Nucleic acid-binding polymers as anti-inflammatory agents: reducing the danger of nuclear attack.

Authors
Pisetsky, DS; Lee, J; Leong, KW; Sullenger, BA
MLA Citation
Pisetsky, DS, Lee, J, Leong, KW, and Sullenger, BA. "Nucleic acid-binding polymers as anti-inflammatory agents: reducing the danger of nuclear attack." Expert Rev Clin Immunol 8.1 (January 2012): 1-3.
PMID
22149331
Source
pubmed
Published In
Expert review of clinical immunology
Volume
8
Issue
1
Publish Date
2012
Start Page
1
End Page
3
DOI
10.1586/eci.11.82

Anti-Müllerian hormone: A better marker of ovarian damage from cyclophosphamide

Authors
Clowse, MEB; Harward, L; Criscione-Schreiber, L; Pisetsky, D; Copland, S
MLA Citation
Clowse, MEB, Harward, L, Criscione-Schreiber, L, Pisetsky, D, and Copland, S. "Anti-Müllerian hormone: A better marker of ovarian damage from cyclophosphamide." Arthritis and Rheumatism 64.5 (2012): 1305-1310.
PMID
22354568
Source
scival
Published In
Arthritis and Rheumatism
Volume
64
Issue
5
Publish Date
2012
Start Page
1305
End Page
1310
DOI
10.1002/art.34431

The inhibition of anti-DNA binding to DNA by nucleic acid binding polymers.

Antibodies to DNA (anti-DNA) are the serological hallmark of systemic lupus erythematosus (SLE) and can mediate disease pathogenesis by the formation of immune complexes. Since blocking immune complex formation can attenuate disease manifestations, the effects of nucleic acid binding polymers (NABPs) on anti-DNA binding in vitro were investigated. The compounds tested included polyamidoamine dendrimer, 1,4-diaminobutane core, generation 3.0 (PAMAM-G3), hexadimethrine bromide, and a β-cylodextrin-containing polycation. As shown with plasma from patients with SLE, NABPs can inhibit anti-DNA antibody binding in ELISA assays. The inhibition was specific since the NABPs did not affect binding to tetanus toxoid or the Sm protein, another lupus autoantigen. Furthermore, the polymers could displace antibody from preformed complexes. Together, these results indicate that NABPs can inhibit the formation of immune complexes and may represent a new approach to treatment.

Authors
Stearns, NA; Lee, J; Leong, KW; Sullenger, BA; Pisetsky, DS
MLA Citation
Stearns, NA, Lee, J, Leong, KW, Sullenger, BA, and Pisetsky, DS. "The inhibition of anti-DNA binding to DNA by nucleic acid binding polymers." PLoS One 7.7 (2012): e40862-.
PMID
22808279
Source
pubmed
Published In
PloS one
Volume
7
Issue
7
Publish Date
2012
Start Page
e40862
DOI
10.1371/journal.pone.0040862

HMGB1 and microparticles as mediators of the immune response to cell death.

In a wide variety of diseases, cell death represents both an outcome and an important step in pathogenesis. This duality occurs because cell death leads to the extracellular release of molecules and structures that can potently induce the innate immune system. These mediators include the alarmins which are endogenous cellular constituents that exit activated or dying cells to stimulate toll-like receptors (TLRs) as well as non-TLR receptors. Of alarmins, the nonhistone protein HMGB1 is the prototype. Like DNA and RNA, HMGB1 can translocate from cells as they die. The activity of HMGB1 may reflect its interaction with other molecules such as LPS, DNA, and cytokines. In addition to alarmins, dead and dying cells can release subcellular organelles called microparticles that contain cytoplasmic and nuclear constituents, including DNA and RNA. These particles can impact on many cell types to induce inflammation. The release of HMGB1 and microparticles shows important similarities, occurring with cell death as well as stimulation of certain but not all TLRs. Furthermore, nitric oxide can induce the release of both. These observations suggest that the products of dead cells can serve as important mediators to drive immune responses and promote inflammation and autoreactivity.

Authors
Pisetsky, DS; Gauley, J; Ullal, AJ
MLA Citation
Pisetsky, DS, Gauley, J, and Ullal, AJ. "HMGB1 and microparticles as mediators of the immune response to cell death." Antioxid Redox Signal 15.8 (October 15, 2011): 2209-2219. (Review)
PMID
21194388
Source
pubmed
Published In
Antioxidants & Redox Signaling
Volume
15
Issue
8
Publish Date
2011
Start Page
2209
End Page
2219
DOI
10.1089/ars.2010.3865

The Association of Intratumoral Germinal Centers with early-stage non-small cell lung cancer.

INTRODUCTION: Lung cancers can display immune cell infiltration although the role of an adaptive immune response in disease pathogenesis is unknown. To investigate the possibility of a functional humoral response to the tumor, we surveyed histologic sections from non-small cell lung cancer (NSCLC) tumors for germinal centers (GCs) and assessed whether there was an association between the presence of GCs and tumor stage. METHODS: Tumor sections from 91 patients with all stages of NSCLC were examined by a pathologist blinded to clinical data. GCs were identified by hematoxylin and eosin staining patterns and confirmed by immunohistochemical staining for B-cell markers, BCL-6 and CD21. The distribution of GCs within the tumor or the tumor margin was recorded. Statistical analysis was performed to evaluate the association between stage and presence of GCs. RESULTS: Thirty-five percent of all tumors evaluated contained GCs, and sections evaluated by immunohistochemistry showed positive staining for both B-cell markers. GCs were seen both within the tumor and the tumor margin, consistent with an immune response to antigen stimulation. Patients with stage I NSCLC had a higher prevalence of intratumoral GCs than patients with stages II to IV (Cochran-Armitage Trend Test p = 0.02011). There was no association of stage with GCs in the tumor margin. CONCLUSIONS: Intratumoral GCs are associated with early-stage NSCLC. Further characterization of intratumoral GCs may lead to new diagnostic and therapeutic strategies based on manipulation of the adaptive immune response.

Authors
Gottlin, EB; Bentley, RC; Campa, MJ; Pisetsky, DS; Herndon, JE; Patz, EF
MLA Citation
Gottlin, EB, Bentley, RC, Campa, MJ, Pisetsky, DS, Herndon, JE, and Patz, EF. "The Association of Intratumoral Germinal Centers with early-stage non-small cell lung cancer." J Thorac Oncol 6.10 (October 2011): 1687-1690.
PMID
21642860
Source
pubmed
Published In
Journal of Thoracic Oncology
Volume
6
Issue
10
Publish Date
2011
Start Page
1687
End Page
1690
DOI
10.1097/JTO.0b013e3182217bec

Identification of novel innate immune genes by transcriptional profiling of macrophages stimulated with TLR ligands.

Toll-like receptors (TLRs) are key receptors in innate immunity and trigger responses following interaction with pathogen-associated molecular patterns (PAMPs). TLR3, TLR4 and TLR9 recognize double stranded RNA, lipopolysaccharide (LPS) and CpG DNA, respectively. These receptors differ importantly in downstream adaptor molecules. TLR4 signals through MyD88 and TRIF; in contrast, the TLR3 pathway involves only TRIF while TLR9 signals solely through MyD88. To determine how differences in downstream signaling could influence gene expression in innate immunity, gene expression patterns were determined for the RAW264.7 macrophage cell line stimulated with LPS, poly (I:C), or CpG DNA. Gene expression profiles 6 and 24h post-stimulation were analyzed to determine genes, pathways and transcriptional networks induced. As these experiments showed, the number and extent of genes expressed varied with stimulus. LPS and poly (I:C) induced an abundant array of genes in RAW264.7 cells at 6h and 24h following treatment while CpG DNA induced many fewer. By analyzing data for networks and pathways, we prioritized differentially expressed genes with respect to those common to the three TLR ligands as well as those shared by LPS and poly (I:C) but not CpG DNA. The importance of changes in gene expression was demonstrated by experiments indicating that RNA interference-mediated inhibition of two genes identified in this analysis, PLEC1 and TPST1, reduced IL-6 production by J774A.1 and RAW264.7 macrophages stimulated with LPS. Together, these findings delineate macrophage gene response patterns induced by different PAMPs and identify new genes that have not previously been implicated in innate immunity.

Authors
Yang, IV; Jiang, W; Rutledge, HR; Lackford, B; Warg, LA; De Arras, L; Alper, S; Schwartz, DA; Pisetsky, DS
MLA Citation
Yang, IV, Jiang, W, Rutledge, HR, Lackford, B, Warg, LA, De Arras, L, Alper, S, Schwartz, DA, and Pisetsky, DS. "Identification of novel innate immune genes by transcriptional profiling of macrophages stimulated with TLR ligands." Mol Immunol 48.15-16 (September 2011): 1886-1895.
PMID
21665277
Source
pubmed
Published In
Molecular Immunology
Volume
48
Issue
15-16
Publish Date
2011
Start Page
1886
End Page
1895
DOI
10.1016/j.molimm.2011.05.015

Effects of progesterone and estradiol sex hormones on the release of microparticles by RAW 264.7 macrophages stimulated by Poly(I:C).

Microparticles (MPs) are small membrane-bound vesicles that display proinflammatory and prothrombotic properties. These particles can be released by macrophages stimulated by ligands of the Toll-like receptors (TLRs) in a process that depends on nitric oxide (NO) production. Since sex hormones can modulate macrophage responses, we investigated the effects of progesterone and estradiol on macrophage particle release in vitro, comparing the responses with those induced by the glucocorticoid dexamethasone. As a model system for particle release, RAW 264.7 cells were stimulated in vitro with poly(I:C), a ligand of TLR3. Microparticles were measured by flow cytometry, while NO was measured by the Griess reaction. As the results of these studies showed, progesterone but not estradiol can block particle release by RAW264.7 cells treated with poly(I:C); dexamethasone was also active. Furthermore, while progesterone and dexamethasone inhibited NO production under the same culture conditions, neither agent blocked the production of particles stimulated by the NO donors dipropylenetriamine NONOate {(z)-1-[N-(3-aminopropyl)-N-(3-ammoniopropyl)amino] diazen-1-ium-1,2-diolate} and (z)-1-[(2-aminoethyl)-N-(2-ammonioethyl)amino] diazen-1-ium-1,2-diolate. Studies using RU486 to assess the role of hormone receptors indicated that while this agent blocked the inhibition of particle and NO production by dexamethasone, it did not affect the inhibition by progesterone. Together, these results indicate that progesterone but not estradiol can inhibit particle release by stimulated macrophages and suggest a mechanism that may contribute to the immunomodulatory effects of this sex hormone.

Authors
Pisetsky, DS; Spencer, DM
MLA Citation
Pisetsky, DS, and Spencer, DM. "Effects of progesterone and estradiol sex hormones on the release of microparticles by RAW 264.7 macrophages stimulated by Poly(I:C)." Clin Vaccine Immunol 18.9 (September 2011): 1420-1426.
PMID
21653747
Source
pubmed
Published In
Clinical and vaccine immunology : CVI
Volume
18
Issue
9
Publish Date
2011
Start Page
1420
End Page
1426
DOI
10.1128/CVI.05110-11

Nucleic acid-binding polymers as anti-inflammatory agents.

Dead and dying cells release nucleic acids. These extracellular RNAs and DNAs can be taken up by inflammatory cells and activate multiple nucleic acid-sensing toll-like receptors (TLR3, 7, 8, and 9). The inappropriate activation of these TLRs can engender a variety of inflammatory and autoimmune diseases. The redundancy of the TLR family encouraged us to seek materials that can neutralize the proinflammatory effects of any nucleic acid regardless of its sequence, structure or chemistry. Herein we demonstrate that certain nucleic acid-binding polymers can inhibit activation of all nucleic acid-sensing TLRs irrespective of whether they recognize ssRNA, dsRNA or hypomethylated DNA. Furthermore, systemic administration of such polymers can prevent fatal liver injury engendered by proinflammatory nucleic acids in an acute toxic shock model in mice. Therefore these polymers represent a novel class of anti-inflammatory agent that can act as molecular scavengers to neutralize the proinflammatory effects of various nucleic acids.

Authors
Lee, J; Sohn, JW; Zhang, Y; Leong, KW; Pisetsky, D; Sullenger, BA
MLA Citation
Lee, J, Sohn, JW, Zhang, Y, Leong, KW, Pisetsky, D, and Sullenger, BA. "Nucleic acid-binding polymers as anti-inflammatory agents." Proc Natl Acad Sci U S A 108.34 (August 23, 2011): 14055-14060.
PMID
21844380
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
108
Issue
34
Publish Date
2011
Start Page
14055
End Page
14060
DOI
10.1073/pnas.1105777108

Laboratory Testing in the Rheumatic Diseases

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Laboratory Testing in the Rheumatic Diseases." 2 (July 1, 2011): 1651-1656. (Chapter)
Source
scopus
Volume
2
Publish Date
2011
Start Page
1651
End Page
1656
DOI
10.1016/B978-1-4377-1604-7.00265-7

Induction of apoptosis in circulating angiogenic cells by microparticles.

OBJECTIVE: Systemic sclerosis (SSc) is an autoimmune disease marked by aberrant activation and apoptosis of endothelial cells (ECs) and decreased numbers of circulating angiogenic cells (CACs). The aim of this study was to analyze whether microparticles might link pathologic activation and apoptosis of ECs with reduced numbers of CACs. METHODS: Apoptosis was quantified by staining for annexin V and measurement of caspase 3 activity. The uptake of microparticles by CACs was determined by fluorescence-activated cell sorting and by fluorescence microscopy. Tritiated arachidonic acid and phosphatidylinositol 3,5-bisphosphate were used to demonstrate the transfer of arachidonic acid and highlight the role of the acid sphingomyelinase in microparticle-induced apoptosis of endothelial progenitor cells. RESULTS: Microparticles derived from activated or apoptotic ECs, the expression of which is strongly increased in the blood of patients with SSc, induce apoptosis in CACs in a dose-dependent manner. Microparticles, which are rich in arachidonic acid, are phagocytosed by CACs. Inhibition of phagocytosis prevents the induction of apoptosis in CACs by microparticles. Microparticles can transport arachidonic acid from ECs to CACs, and purified arachidonic acid mimics the proapoptotic effects of microparticles. Arachidonic acid activates the acid sphingomyelinase, and inhibition of acid sphingomyelinase prevents microparticle-induced apoptosis of CACs. Thus, phagocytosis of microparticles might stimulate the activity of acid sphingomyelinase and activate the apoptotic machinery. CONCLUSION: The induction of apoptosis in CACs by microparticles derived from ECs provides a novel link between aberrant activation or apoptosis of ECs, decreased numbers of CACs, and impaired formation of new vessels in SSc.

Authors
Distler, JHW; Akhmetshina, A; Dees, C; Jüngel, A; Stürzl, M; Gay, S; Pisetsky, DS; Schett, G; Distler, O
MLA Citation
Distler, JHW, Akhmetshina, A, Dees, C, Jüngel, A, Stürzl, M, Gay, S, Pisetsky, DS, Schett, G, and Distler, O. "Induction of apoptosis in circulating angiogenic cells by microparticles." Arthritis Rheum 63.7 (July 2011): 2067-2077.
PMID
21437873
Source
pubmed
Published In
Arthritis and Rheumatism
Volume
63
Issue
7
Publish Date
2011
Start Page
2067
End Page
2077
DOI
10.1002/art.30361

Microparticles as antigenic targets of antibodies to DNA and nucleosomes in systemic lupus erythematosus.

Systemic lupus erythematosus is a prototypic autoimmune disease characterized by antibodies to DNA and other nuclear molecules. While these antibodies can form immune complexes, the mechanisms generating the bound nuclear antigens are not known. These studies investigated whether microparticles can form complexes with anti-DNA and other anti-nucleosomal antibodies. Microparticles are small membrane-bound vesicles released from dead and dying cells; these particles contain a variety of cellular components, including DNA. To assess antigenicity, microparticles generated in vitro from apoptotic cell lines were tested using murine monoclonal anti-DNA and anti-nucleosomal antibodies as well as plasma from lupus patients. Antibody binding was assessed by flow cytometry. As these studies showed, some but not all of the monoclonal antibodies bound to microparticles prepared from apoptotic HL-60, THP-1 and Jurkat cells. For HL-60 cells, both staurosporine and UV radiation led to the production of antigenically active particles. For the anti-DNA antibody with high particle binding, prior treatment of DNase reduced activity. With plasma from patients with SLE, antibody binding to microparticles was present although a clear relationship with anti-DNA antibody levels was not observed. To determine whether lupus plasma contains immune complexes with particle properties, particle preparations were tested for bound IgG by flow cytometry. These studies indicated that lupus plasma contains particles with IgG binding, with numbers correlated with anti-DNA levels. Together, these findings indicate that microparticles display DNA and nucleosomal molecules in an antigenic form and could represent a source of immune complexes in SLE.

Authors
Ullal, AJ; Reich, CF; Clowse, M; Criscione-Schreiber, LG; Tochacek, M; Monestier, M; Pisetsky, DS
MLA Citation
Ullal, AJ, Reich, CF, Clowse, M, Criscione-Schreiber, LG, Tochacek, M, Monestier, M, and Pisetsky, DS. "Microparticles as antigenic targets of antibodies to DNA and nucleosomes in systemic lupus erythematosus." J Autoimmun 36.3-4 (May 2011): 173-180.
PMID
21376534
Source
pubmed
Published In
Journal of Autoimmunity
Volume
36
Issue
3-4
Publish Date
2011
Start Page
173
End Page
180
DOI
10.1016/j.jaut.2011.02.001

Antinuclear antibodies in healthy people: the tip of autoimmunity's iceberg?

Antinuclear antibodies (ANAs) are venerable biomarkers for assessing the diagnosis and prognosis of patients with autoimmunity. While closely associated with diseases such as systemic lupus erythematosus, ANA expression occurs commonly in healthy people. The basis for this expression is unknown, although it may reflect features of the assays for antibody detection or intrinsic immunological disturbances in otherwise normal individuals. Like autoimmunity itself, ANA expression is more common among women than men, pointing to an important determinant of these responses. Future research will clarify the mechanisms of ANA expression and the utility of current assays as antecedent and screening biomarkers.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Antinuclear antibodies in healthy people: the tip of autoimmunity's iceberg? (Published online)." Arthritis Res Ther 13.2 (April 21, 2011): 109-.
PMID
21554754
Source
pubmed
Published In
Arthritis Research and Therapy
Volume
13
Issue
2
Publish Date
2011
Start Page
109
DOI
10.1186/ar3282

Microparticles as a source of extracellular DNA.

Microparticles are small membrane-bound vesicles that display pro-inflammatory and pro-thrombotic activities important in the pathogenesis of a wide variety of diseases. These particles are released from activated and dying cells and incorporate nuclear and cytoplasmic molecules for extracellular export. Of these molecules, DNA is a central autoantigen in systemic lupus erythematosus (SLE). As studies in our laboratory show, DNA occurs prominently in microparticles, translocating into these structures during apoptotic cell death. This DNA is antigenically active and can bind to lupus anti-DNA autoantibodies. These findings suggest that microparticles are an important source of extracellular DNA to serve as an autoantigen and autoadjuvant in SLE.

Authors
Pisetsky, DS; Gauley, J; Ullal, AJ
MLA Citation
Pisetsky, DS, Gauley, J, and Ullal, AJ. "Microparticles as a source of extracellular DNA." Immunol Res 49.1-3 (April 2011): 227-234.
PMID
21132466
Source
pubmed
Published In
Immunologic Research
Volume
49
Issue
1-3
Publish Date
2011
Start Page
227
End Page
234
DOI
10.1007/s12026-010-8184-8

Platelet microparticles: making blood a bad humor.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Platelet microparticles: making blood a bad humor." J Rheumatol 38.4 (April 2011): 590-592.
PMID
21459954
Source
pubmed
Published In
The Journal of rheumatology
Volume
38
Issue
4
Publish Date
2011
Start Page
590
End Page
592
DOI
10.3899/jrheum.101359

Gamow, guppies, and the search for GOD

© Springer Science+Business Media B.V. 2011. The term physician-scientist is one of those compound words that have been created to unite disparate elements. Our language has others: student-athlete, warrior-statesman, and player-coach. The hyphen is a convenient way to keep the words together, but the hyphen cannot obscure the inherent contradictions that fight within. At that core, physicians and scientists (just like scholars and athletes) are worlds apart. Becoming a physician-scientist demands a union that can take years to forge and is often tenuous and unnerving.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Gamow, guppies, and the search for GOD." Medicine Science and Dreams: The Making of Physician-Scientists. January 1, 2011. 15-32.
Source
scopus
Publish Date
2011
Start Page
15
End Page
32
DOI
10.1007/978-90-481-9538-1_2

The 2011 ACR meeting: what's new in the Windy City.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "The 2011 ACR meeting: what's new in the Windy City." Arthritis Res Ther 13.6 (2011): 138-.
PMID
22192758
Source
pubmed
Published In
Arthritis Research and Therapy
Volume
13
Issue
6
Publish Date
2011
Start Page
138
DOI
10.1186/ar3532

Pus: the Rodney Dangerfield of immunology.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Pus: the Rodney Dangerfield of immunology." Arthritis Res Ther 13.5 (2011): 131-.
PMID
22029909
Source
pubmed
Published In
Arthritis Research and Therapy
Volume
13
Issue
5
Publish Date
2011
Start Page
131
DOI
10.1186/ar3477

Cell death in the pathogenesis of immune-mediated diseases: the role of HMGB1 and DAMP-PAMP complexes.

Cell death is a ubiquitous process whose immunological consequences can influence the course of infectious, autoimmune and inflammatory diseases. While cell death has long been dichotomised in terms of apoptosis and necrosis, other forms of death can occur and they vary in their capacity to stimulate as well as inhibit inflammation. The pro-inflammatory activity of dead cells results from a wide variety of intracellular molecules that are released as cell permeability increases during death. These molecules have been termed as DAMPs (damage associated molecular patterns) or alarmins. Among these DAMPs, HMGB1, a non-histone nuclear protein, serves as the prototype. Although HMGB1 was originally thought to act alone as a cytokine, recent studies suggest that its immunological effects result from complexes of HMGB1 with either other DAMPs or with PAMPs (pathogen associated molecular patterns). Studies on the role of HMGB1 in pathogenesis suggest that the formation of extracellular complexes is an important mechanism for generating pro-inflammatory signals during cell death and therefore could be a potential target of new therapy.

Authors
Pisetsky, D
MLA Citation
Pisetsky, D. "Cell death in the pathogenesis of immune-mediated diseases: the role of HMGB1 and DAMP-PAMP complexes." Swiss medical weekly 141 (2011): w13256-.
PMID
21877298
Source
scival
Published In
Swiss medical weekly : official journal of the Swiss Society of Infectious Diseases, the Swiss Society of Internal Medicine, the Swiss Society of Pneumology
Volume
141
Publish Date
2011
Start Page
w13256
DOI
10.4414/smw.2011.13256

Pus: the Rodney Dangerfield of immunology.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Pus: the Rodney Dangerfield of immunology." Arthritis research & therapy 13.5 (2011): 131--.
Source
scival
Published In
Arthritis Research and Therapy
Volume
13
Issue
5
Publish Date
2011
Start Page
131-
DOI
10.1186/ar3477

The role of HMGB1 in efferocytosis: when the dead go unburied. Focus on "HMGB1 inhibits macrophage activity in efferocytosis through binding to the alphavbeta3-integrin".

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "The role of HMGB1 in efferocytosis: when the dead go unburied. Focus on "HMGB1 inhibits macrophage activity in efferocytosis through binding to the alphavbeta3-integrin"." Am J Physiol Cell Physiol 299.6 (December 2010): C1253-C1255.
PMID
20926774
Source
pubmed
Published In
American journal of physiology. Cell physiology
Volume
299
Issue
6
Publish Date
2010
Start Page
C1253
End Page
C1255
DOI
10.1152/ajpcell.00397.2010

The blood nucleome in the pathogenesis of SLE.

Systemic lupus erythematosus (SLE) is prototypic autoimmune disease characterized by the production of autoantibodies to DNA among other nuclear molecules. These antibodies can form immune complexes that promote pathogenesis by stimulating cytokine production and depositing in the kidney to instigate nephritis. The antigens that form these complexes arise from the blood nucleome, a pool of circulating macromolecules comprised of DNA, RNA and nuclear proteins released from cells. Cell death is a major source of these molecules, releasing DNA in a process that can be modeled in mice by the administration of cells killed ex vivo. In the mouse model, the appearance of blood DNA requires macrophages and differs between males and females. This finding raises the possibility that augmented levels of extracellular DNA and other nuclear antigens can contribute to the increased frequency of SLE in females. Extracellular DNA can occur in both a soluble and particulate form, with microparticles generated in vitro displaying antigenically active DNA. Together, these findings suggest that cell death is an important event in lupus pathogenesis and can provide a supply of blood DNA essential for immune complex formation.

Authors
Pisetsky, DS; Ullal, AJ
MLA Citation
Pisetsky, DS, and Ullal, AJ. "The blood nucleome in the pathogenesis of SLE." Autoimmun Rev 10.1 (November 2010): 35-37. (Review)
PMID
20659590
Source
pubmed
Published In
Autoimmunity Reviews
Volume
10
Issue
1
Publish Date
2010
Start Page
35
End Page
37
DOI
10.1016/j.autrev.2010.07.007

Charlie's List.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Charlie's List." Ann Intern Med 153.5 (September 7, 2010): 344-.
PMID
20820046
Source
pubmed
Published In
Annals of internal medicine
Volume
153
Issue
5
Publish Date
2010
Start Page
344
DOI
10.7326/0003-4819-153-5-201009070-00012

The release of microparticles by RAW 264.7 macrophage cells stimulated with TLR ligands.

MPs are small membrane-bound particles that originate from activated and dying cells and mediate intercellular communication. Once released from cells, MPs can serve as novel signaling elements in innate immunity, with levels elevated in immune-mediated diseases. This study tested the hypothesis that TLR stimulation can induce MP release by macrophages. In these experiments, using the RAW 264.7 murine macrophage cell line as a model, LPS, a TLR4 ligand, and poly(I:C), a TLR3 ligand, induced MP release effectively, as measured by flow cytometry; in contrast, a CpG oligonucleotide, which can stimulate TLR9, induced much lower levels of particle release. To determine the role of other mediators in this response, the effects of NO were tested. Thus, MP release from RAW 264.7 cells stimulated by LPS or poly(I:C) correlated with NO production, and treatment with the iNOS inhibitor 1400W decreased particle release and NO production. Furthermore, treatment of RAW 264.7 cells with NO donors induced MP production. As TLR ligands can induce apoptosis, the effect of caspase inhibition on MP release by stimulated cells was assessed. These experiments showed that the pan-caspase inhibitor, ZVAD, although decreasing NO production, increased MP release by stimulated cells. Together, these experiments demonstrate that TLR stimulation of macrophages can lead to MP release, and NO plays a key role in this response.

Authors
Gauley, J; Pisetsky, DS
MLA Citation
Gauley, J, and Pisetsky, DS. "The release of microparticles by RAW 264.7 macrophage cells stimulated with TLR ligands." J Leukoc Biol 87.6 (June 2010): 1115-1123.
PMID
20335312
Source
pubmed
Published In
Journal of leukocyte biology
Volume
87
Issue
6
Publish Date
2010
Start Page
1115
End Page
1123
DOI
10.1189/jlb.0709465

The release of microparticles by Jurkat leukemia T cells treated with staurosporine and related kinase inhibitors to induce apoptosis.

Microparticles (MPs) are small membrane-bound vesicles released from cells undergoing activation or cell death. These particles display potent biological activities that can impact on physiologic and pathologic processes. Previous studies with the Jurkat T leukemia cell line demonstrated that staurosporine (STS) induces the release of MPs as cells undergo apoptosis. To investigate further this process, we tested the effects of STS, its analogue, 7-hydroxystaurosporine (UCN-01), and other protein kinase C (PKC) and cyclin-dependent kinase (CDK) inhibitors. FACS analysis was used to assess MP release. Results of these studies indicate that STS and UCN-01 induce MP release by Jurkat cells; in contrast, other PKC and CDK inhibitors failed to induce comparable release, suggesting that release does not result from simple inhibition of either kinase alone. Time course experiments indicated that STS-induced particle release occurred as early as 2 h after treatment, with the early release MPs displaying low levels of binding of annexin V and propidium iodide (PI). Early-release MPs, however, matured in culture to an annexin V- and PI-positive phenotype. Together, these results indicate that STS and UCN-01 induce MPs that are phenotypically distinct and reflect specific patterns of kinase inhibition during apoptosis.

Authors
Ullal, AJ; Pisetsky, DS
MLA Citation
Ullal, AJ, and Pisetsky, DS. "The release of microparticles by Jurkat leukemia T cells treated with staurosporine and related kinase inhibitors to induce apoptosis." Apoptosis 15.5 (May 2010): 586-596.
PMID
20146001
Source
pubmed
Published In
Apoptosis
Volume
15
Issue
5
Publish Date
2010
Start Page
586
End Page
596
DOI
10.1007/s10495-010-0470-3

HMGB1: a dangerous player in lupus pathogenesis.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "HMGB1: a dangerous player in lupus pathogenesis." J Rheumatol 37.4 (April 2010): 689-691.
PMID
20360203
Source
pubmed
Published In
The Journal of rheumatology
Volume
37
Issue
4
Publish Date
2010
Start Page
689
End Page
691
DOI
10.3899/jrheum.091459

Use of SYTO 13, a fluorescent dye binding nucleic acids, for the detection of microparticles in in vitro systems.

Microparticles (MPs) are small membrane-bound vesicles that are released from activated or dying cells by a blebbing process. These particles contain nuclear and cytoplasmic components and represent unique biomarkers for disease. The small size of particles, however, limits detection using flow cytometry with either light scatter or staining for surface markers. Because MPs contain DNA and RNA, we have explored the use of SYTO 13, a member of the class of SYTO dyes, for particle detection. SYTO 13 is cell permeable and has a high fluorescent yield when bound to DNA or RNA. In this study, we compared detection of MPs using either light scatter or SYTO 13 staining, testing the hypothesis that, with fluorescence detection with SYTO 13, problems of "noise" with light scatter are reduced and the range of MP sizes detected is increased. In these experiments, particles were obtained from lymphoid cell lines treated in vitro to undergo apoptosis. As these results showed, STYO 13 allowed the detection of 1.5-2.9 times as many particles as did light scatter. The increased sensitivity was observed with three different cell lines and was independent of inducing stimulus. Treatment of fixed and permeabilized MPs with DNase and RNase showed that SYTO 13 binding resulted from interaction with both DNA and RNA. Together, these findings indicate that the nucleic acid content of MPs provides the basis for their detection in in vitro systems and suggests the utility of fluorescent dyes like SYTO 13 for more sensitive quantitative assays.

Authors
Ullal, AJ; Pisetsky, DS; Reich, CF
MLA Citation
Ullal, AJ, Pisetsky, DS, and Reich, CF. "Use of SYTO 13, a fluorescent dye binding nucleic acids, for the detection of microparticles in in vitro systems." Cytometry A 77.3 (March 2010): 294-301.
PMID
20104574
Source
pubmed
Published In
Cytometry
Volume
77
Issue
3
Publish Date
2010
Start Page
294
End Page
301
DOI
10.1002/cyto.a.20833

Effects of HMGB1 on in vitro responses of isolated muscle fibers and functional aspects in skeletal muscles of idiopathic inflammatory myopathies.

Idiopathic inflammatory myopathies (IIMs) are heterogeneous rheumatic disorders of unknown cause characterized by muscle weakness, inflammatory cell infiltrates, and major histocompatibility complex (MHC) class I expression on muscle fibers. The nonhistone nuclear protein alarmin high-mobility group box 1 protein (HMGB1) has been detected extranuclearly in muscle biopsies from patients with IIMs. We hypothesize that HMGB1 has a central role in the cause of muscle weakness, particularly in the early phases of IIMs. Experiments were performed on skeletal muscle fibers isolated from adult mice, which were exposed to recombinant interferon (IFN)-gamma or HMGB1. The myoplasmic free [Ca(2+)] was measured. Stimulation with IFN-gamma resulted in increased HMGB1 expression in muscle nuclei and the myoplasm. Exposure to HMGB1 induced a reversible up-regulation of MHC class I in the muscle fibers. However, HMGB1 exposure caused an irreversible decrease in Ca(2+) release from the sarcoplasmic reticulum during fatigue, induced by repeated tetanic contractions. HMGB1 and MHC class I were frequently colocalized in the myoplasm of muscle fibers in muscle biopsies from patients with early IIMs. However, HMGB1-expressing fibers outnumbered fibers expressing MHC class I. Our data indicate that HMGB1 could be an early inducer of skeletal muscle dysfunction in IIMs.

Authors
Grundtman, C; Bruton, J; Yamada, T; Ostberg, T; Pisetsky, DS; Harris, HE; Andersson, U; Lundberg, IE; Westerblad, H
MLA Citation
Grundtman, C, Bruton, J, Yamada, T, Ostberg, T, Pisetsky, DS, Harris, HE, Andersson, U, Lundberg, IE, and Westerblad, H. "Effects of HMGB1 on in vitro responses of isolated muscle fibers and functional aspects in skeletal muscles of idiopathic inflammatory myopathies." FASEB J 24.2 (February 2010): 570-578.
PMID
19837864
Source
pubmed
Published In
The FASEB journal : official publication of the Federation of American Societies for Experimental Biology
Volume
24
Issue
2
Publish Date
2010
Start Page
570
End Page
578
DOI
10.1096/fj.09-144782

The role of microparticles in the pathogenesis of rheumatic diseases.

Microparticles (MPs) are small membrane-bound vesicles that are emerging as important elements in the pathogenesis of rheumatic diseases owing to their pleiotropic effects on thrombosis, vascular reactivity, angiogenesis and inflammation. Released from cells during activation and apoptosis, MPs carry proteins, lipids and nucleic acids, and serve as platforms for enzymatic processes in thrombosis. Furthermore, MPs can transfer cytokines, receptors, RNA and DNA to modulate the properties of target cells. As MPs appear in the blood in increased numbers during rheumatic disease, they represent novel biomarkers that can be used to assess events in otherwise inaccessible tissues. Future research will define further the pathogenetic role of MPs and explore therapeutic strategies to block their release or signaling properties.

Authors
Beyer, C; Pisetsky, DS
MLA Citation
Beyer, C, and Pisetsky, DS. "The role of microparticles in the pathogenesis of rheumatic diseases." Nat Rev Rheumatol 6.1 (January 2010): 21-29. (Review)
PMID
19949432
Source
pubmed
Published In
Nature Reviews Rheumatology
Volume
6
Issue
1
Publish Date
2010
Start Page
21
End Page
29
DOI
10.1038/nrrheum.2009.229

Report of the American College of Rheumatology Pain Management Task Force

The next few years in medicine will be a time of incredible change, and the organizational structure (e.g., task force) overseeing the pain initiative should regularly monitor the activities of the initiative, survey members, and report back to the BOD. The initiative has an impact on the identity of the specialty of rheumatology and it is essential that it is supported by the members and has the resources, both intellectual and financial, to assure its success. © 2010, American College of Rheumatology.

Authors
Borenstein, D; Altman, R; Bello, A; Chatham, W; Clauw, D; Crofford, L; Croft, J; Hassett, A; Kozin, F; Pisetsky, D; Richardson, J; Schanberg, L; Starz, T; Witter, J
MLA Citation
Borenstein, D, Altman, R, Bello, A, Chatham, W, Clauw, D, Crofford, L, Croft, J, Hassett, A, Kozin, F, Pisetsky, D, Richardson, J, Schanberg, L, Starz, T, and Witter, J. "Report of the American College of Rheumatology Pain Management Task Force." Arthritis Care and Research 62.5 (2010): 590-599.
PMID
20461782
Source
scival
Published In
Arthritis Care and Research
Volume
62
Issue
5
Publish Date
2010
Start Page
590
End Page
599
DOI
10.1002/acr.20005

Antibodies to DNA: infection or genetics?

Antibodies to DNA (anti-DNA) are the serological hallmark of systemic lupus erythematosus (SLE) and unique markers of the immunological disturbances critical to disease pathogenesis. In the form of immune complexes, anti-DNA autoantibodies can deposit in the tissue to incite inflammation and damage; in addition, these complexes can induce cytokine production, most prominently, type 1 interferon. Studies in both patients and animal models have implicated genetic as well as environmental factors in the aetiology of the anti-DNA response. Because bacterial DNA is a potent stimulant of innate immunity by both toll-like receptor (TLR) and non-TLR signalling pathways, foreign DNA introduced during the course of bacterial or viral infection could have a dual role in antibody induction. This DNA could serve as an adjuvant to activate innate immunity as well as an immunogen to drive an antigen-specific antibody response. In this scenario, the generation of cross-reactive autoantibodies, in contrast to highly specific antibodies to bacterial DNA, most likely depends on genetically determined abnormalities in the B-cell repertoire in patients with SLE. Given the universal expression of DNA, this model suggests that many different kinds of infections could trigger pathogenic autoantibody responses in SLE, as well as induce flare.

Authors
Pisetsky, DS; Vrabie, IA
MLA Citation
Pisetsky, DS, and Vrabie, IA. "Antibodies to DNA: infection or genetics?." Lupus 18.13 (November 2009): 1176-1180. (Review)
PMID
19880564
Source
pubmed
Published In
Lupus
Volume
18
Issue
13
Publish Date
2009
Start Page
1176
End Page
1180
DOI
10.1177/0961203309106492

The role of extracellular DNA in autoimmunity in SLE.

Systemic lupus erythematosus (SLE) is a prototypic autoimmune disease characterized by the expression of antibodies to DNA. These antibodies form immune complexes that can stimulate cytokine production as well as deposit in the tissues to incite inflammation and damage. For the formation of immune complexes, the availability of extracellular DNA in an immunologically relevant form is essential. While apoptosis has been implicated as the source of this nuclear material in SLE, as shown with in vitro or in vivo systems, extracellular DNA can originate from apoptotic as well as necrotic cells. In experimental models, the release of DNA occurs with the administration of cells induced to die, in vitro as well as the administration of agents to induce cell death in situ. This release can be influenced by the presence of inflammatory cells such as macrophages that can interact with dead and dying cells to modulate the translocation of DNA from the inside to the outside of cells. In vivo, both glucocorticoids and oestrogens can modify the extent of DNA release from the administration of dead and dying cells. Together, these findings indicate that the generation of extracellular DNA in SLE can result from cell death and that steps in this process represent potential targets for new therapies.

Authors
Su, K-Y; Pisetsky, DS
MLA Citation
Su, K-Y, and Pisetsky, DS. "The role of extracellular DNA in autoimmunity in SLE." Scand J Immunol 70.3 (September 2009): 175-183. (Review)
PMID
19703007
Source
pubmed
Published In
Scandinavian Journal of Immunology
Volume
70
Issue
3
Publish Date
2009
Start Page
175
End Page
183
DOI
10.1111/j.1365-3083.2009.02300.x

Systemic lupus erythematosus: a matter of life and death.

Authors
Pisetsky, DS; Rönnblom, L
MLA Citation
Pisetsky, DS, and Rönnblom, L. "Systemic lupus erythematosus: a matter of life and death." Arthritis Rheum 60.6 (June 2009): 1567-1570.
PMID
19479835
Source
pubmed
Published In
Arthritis and Rheumatism
Volume
60
Issue
6
Publish Date
2009
Start Page
1567
End Page
1570
DOI
10.1002/art.24531

The translocation of HMGB1 during cell activation and cell death.

High-mobility group box protein 1 (HMGB1) is a non-histone nuclear protein with alarmin activity. When present in an extracellular location, HMGB1 can activate the innate immune system and promote inflammation in conditions such as sepsis. To exert these activities, HMGB1 must transit from the nucleus, through the cytoplasm, to the outside of the cell. This process can occur during cell activation as well as cell death. In murine macrophages (MPhi), stimulation of TLR3 and TLR4, but not TLR9, can cause HMGB1 translocation. With cell death, necrosis can lead to extracellular HMGB1 by a passive mechanism. With apoptosis, HMGB1 is only released during secondary necrosis, when cell permeability barriers break down. Since agents that stimulate MPhi can also induce apoptosis, HMGB1 release following TLR stimulation may also reflect a contribution from dead cells, suggesting a common mechanism for protein release in activation and death.

Authors
Gauley, J; Pisetsky, DS
MLA Citation
Gauley, J, and Pisetsky, DS. "The translocation of HMGB1 during cell activation and cell death." Autoimmunity 42.4 (May 2009): 299-301. (Review)
PMID
19811282
Source
pubmed
Published In
Autoimmunity (Informa)
Volume
42
Issue
4
Publish Date
2009
Start Page
299
End Page
301

Post-Translational Modification of HMGB1 and Its Role in Immune Activation

High-mobility group box 1 (HMGB1) functions as an alarmin following release from activated or necrotic cells and mediates inflammation. Translocation and extracellular release ofHMGB1can also occur during apoptosis. This translocation ofHMGB1from the nucleus to the cytoplasm results from post-translational modifications similar to those affecting histones. Such post-translational modifications of HMGB1 could also have effects on gene expression following changes in its DNA-binding properties. Furthermore, modified HMGB1 in the extracellular environment displays immunological activity and could serve as a potential target for new therapy. © 2009 John Wiley & Sons Ltd.

Authors
Ullal, AJ; Pisetsky, DS
MLA Citation
Ullal, AJ, and Pisetsky, DS. "Post-Translational Modification of HMGB1 and Its Role in Immune Activation." (April 29, 2009): 165-178. (Chapter)
Source
scopus
Publish Date
2009
Start Page
165
End Page
178
DOI
10.1002/9780470743553.ch10

Glomerular type 1 angiotensin receptors augment kidney injury and inflammation in murine autoimmune nephritis.

Studies in humans and animal models indicate a key contribution of angiotensin II to the pathogenesis of glomerular diseases. To examine the role of type 1 angiotensin (AT1) receptors in glomerular inflammation associated with autoimmune disease, we generated MRL-Faslpr/lpr (lpr) mice lacking the major murine type 1 angiotensin receptor (AT1A); lpr mice develop a generalized autoimmune disease with glomerulonephritis that resembles SLE. Surprisingly, AT1A deficiency was not protective against disease but instead substantially accelerated mortality, proteinuria, and kidney pathology. Increased disease severity was not a direct effect of immune cells, since transplantation of AT1A-deficient bone marrow did not affect survival. Moreover, autoimmune injury in extrarenal tissues, including skin, heart, and joints, was unaffected by AT1A deficiency. In murine systems, there is a second type 1 angiotensin receptor isoform, AT1B, and its expression is especially prominent in the renal glomerulus within podocytes. Further, expression of renin was enhanced in kidneys of AT1A-deficient lpr mice, and they showed evidence of exaggerated AT1B receptor activation, including substantially increased podocyte injury and expression of inflammatory mediators. Administration of losartan, which blocks all type 1 angiotensin receptors, reduced markers of kidney disease, including proteinuria, glomerular pathology, and cytokine mRNA expression. Since AT1A-deficient lpr mice had low blood pressure, these findings suggest that activation of type 1 angiotensin receptors in the glomerulus is sufficient to accelerate renal injury and inflammation in the absence of hypertension.

Authors
Crowley, SD; Vasievich, MP; Ruiz, P; Gould, SK; Parsons, KK; Pazmino, AK; Facemire, C; Chen, BJ; Kim, H-S; Tran, TT; Pisetsky, DS; Barisoni, L; Prieto-Carrasquero, MC; Jeansson, M; Foster, MH; Coffman, TM
MLA Citation
Crowley, SD, Vasievich, MP, Ruiz, P, Gould, SK, Parsons, KK, Pazmino, AK, Facemire, C, Chen, BJ, Kim, H-S, Tran, TT, Pisetsky, DS, Barisoni, L, Prieto-Carrasquero, MC, Jeansson, M, Foster, MH, and Coffman, TM. "Glomerular type 1 angiotensin receptors augment kidney injury and inflammation in murine autoimmune nephritis." J Clin Invest 119.4 (April 2009): 943-953.
PMID
19287096
Source
pubmed
Published In
Journal of Clinical Investigation
Volume
119
Issue
4
Publish Date
2009
Start Page
943
End Page
953
DOI
10.1172/JCI34862

The content of DNA and RNA in microparticles released by Jurkat and HL-60 cells undergoing in vitro apoptosis.

Microparticles are small membrane-bound vesicles that are released from apoptotic cells during blebbing. These particles contain DNA and RNA and display important functional activities, including immune system activation. Furthermore, nucleic acids inside the particle can be analyzed as biomarkers in a variety of disease states. To elucidate the nature of microparticle nucleic acids, DNA and RNA released in microparticles from the Jurkat T and HL-60 promyelocytic cell lines undergoing apoptosis in vitro were studied. Microparticles were isolated from culture media by differential centrifugation and characterized by flow cytometry and molecular approaches. In these particles, DNA showed laddering by gel electrophoresis and was present in a form that allowed direct binding by a monoclonal anti-DNA antibody, suggesting antigen accessibility even without fixation. Analysis of RNA by gel electrophoresis showed intact 18s and 28s ribosomal RNA bands, although lower molecular bands consistent with 28s ribosomal RNA degradation products were also present. Particles also contained messenger RNA as shown by RT-PCR amplification of sequences for beta-actin and GAPDH. In addition, gel electrophoresis showed the presence of low molecular weight RNA in the size range of microRNA. Together, these results indicate that microparticles from apoptotic Jurkat and HL-60 cells contain diverse nucleic acid species, indicating translocation of both nuclear and cytoplasmic DNA and RNA as particle release occurs during death.

Authors
Reich, CF; Pisetsky, DS
MLA Citation
Reich, CF, and Pisetsky, DS. "The content of DNA and RNA in microparticles released by Jurkat and HL-60 cells undergoing in vitro apoptosis." Exp Cell Res 315.5 (March 10, 2009): 760-768.
PMID
19146850
Source
pubmed
Published In
Experimental Cell Research
Volume
315
Issue
5
Publish Date
2009
Start Page
760
End Page
768
DOI
10.1016/j.yexcr.2008.12.014

Microparticles as biomarkers in autoimmunity: from dust bin to center stage.

Microparticles are small membrane-bound vesicles released from activated and dying cells. As shown in a study of primary Sjogren's syndrome, systemic lupus erythematosus and rheumatoid arthritis, levels of microparticles in the blood, as measured by a solid-phase prothrombinase assay or flow cytometry, are increased with autoimmunity. Among patients with these conditions, however, particle numbers were inversely related to disease activity and levels of the enzyme secretory phospholipase A2 that can digest membrane lipids and perhaps cause particle loss. These findings suggest microparticles as novel biomarkers for autoimmunity, with levels reflecting events leading to their loss as well as production.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Microparticles as biomarkers in autoimmunity: from dust bin to center stage." Arthritis Res Ther 11.6 (2009): 135-.
PMID
19954508
Source
pubmed
Published In
Arthritis Research and Therapy
Volume
11
Issue
6
Publish Date
2009
Start Page
135
DOI
10.1186/ar2856

Pain in rheumatoid arthritis and osteoarthritis

Authors
Pisetsky, DS; McCleane, G
MLA Citation
Pisetsky, DS, and McCleane, G. "Pain in rheumatoid arthritis and osteoarthritis." Pain Management Secrets (2009): 170-183.
Source
scival
Published In
Pain Management Secrets
Publish Date
2009
Start Page
170
End Page
183
DOI
10.1016/B978-0-323-04019-8.00027-5

Systemic lupus erythematosus B. epidemiology, pathology, and pathogenesis

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Systemic lupus erythematosus B. epidemiology, pathology, and pathogenesis." (December 1, 2008): 319-326. (Chapter)
Source
scopus
Publish Date
2008
Start Page
319
End Page
326
DOI
10.1007/978-0-387-68566-3_15

The role of innate immunity in the induction of autoimmunity.

The autoimmune diseases are a diverse group of conditions characterized by abnormal B and T cell reactivity in association with autoantibody production. Among these diseases, systemic lupus erythematosus (SLE) is notable for the expression of antibodies to DNA, with these antibodies representing diagnostic markers. While mammalian DNA is immunologically inert, DNA from bacteria can potently stimulate the innate immune system, activating both toll-like receptors (TLRs) as well as non-TLR internal receptors. Since the sera of normal humans contain antibodies specific for bacterial DNA, this molecule appears to be immunogenic during infection. In pre-autoimmune mice, immunization with bacterial DNA can induce anti-DNA autoantibody production, suggesting a role in initiating this response. The immune properties of DNA are mutable, however, since mammalian DNA can acquire immunological activity when bound to certain proteins or anti-DNA antibodies to form immune complexes. In SLE, these immune complexes can drive the production of interferon by plasmacytoid dendritic cells, thereby intensifying autoimmunity. Together, these observations suggest that DNA can induce innate as well as adaptive immune responses and promote the pathogenesis of SLE because of its intrinsic immunostimulatory activity.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "The role of innate immunity in the induction of autoimmunity." Autoimmun Rev 8.1 (October 2008): 69-72. (Review)
PMID
18708168
Source
pubmed
Published In
Autoimmunity Reviews
Volume
8
Issue
1
Publish Date
2008
Start Page
69
End Page
72
DOI
10.1016/j.autrev.2008.07.028

The relationship between plasma microparticles and disease manifestations in patients with systemic sclerosis.

OBJECTIVE: Microparticles are small, membrane-coated vesicles that can serve as novel signaling structures between cells. The aim of the present study was to analyze the profile of microparticles in the blood of patients with systemic sclerosis (SSc; scleroderma) and healthy controls. METHODS: The study population consisted of 37 patients with SSc and 15 healthy subjects of comparable sex and age. Microparticles were isolated from plasma by high-speed differential centrifugation. Microparticles were stained with monoclonal antibodies against cell type-specific markers and were quantified by fluorescence-activated cell sorting analyses. RESULTS: The total number of microparticles was strongly increased in patients with SSc compared with healthy controls (mean +/- SEM 88.0 +/- 4.8 x 10(5) microparticles/ml plasma versus 42.3 +/- 9.4 x 10(5) microparticles/ml plasma; P < 0.001). Similarly, significant increases were found for microparticles derived from platelets, endothelial cells, monocytes, and T cells, reflecting the activation of these cells in SSc. Platelets were the most common source of microparticles in the blood of patients with SSc (66.9 +/- 5.2% of all microparticles) and healthy donors, followed by microparticles derived from endothelial cells (8.8 +/- 0.9% in SSc patients). The modified Rodnan skin thickness score (MRSS) was inversely correlated with the total number of microparticles. Furthermore, patients with cutaneous ulcers showed a significantly lower total number of microparticles. In multivariate analysis, an additive model of age, C-reactive protein, MRSS, and subtype of disease accounted for 55% of the variability of the total microparticle count (r = 0.744). CONCLUSION: The number of microparticles from different cellular sources is increased in the blood of SSc patients. Considering their role as important mediators of intercellular communication, microparticles could be a novel link between activated cellular compartments in the pathogenesis of SSc.

Authors
Guiducci, S; Distler, JHW; Jüngel, A; Huscher, D; Huber, LC; Michel, BA; Gay, RE; Pisetsky, DS; Gay, S; Matucci-Cerinic, M; Distler, O
MLA Citation
Guiducci, S, Distler, JHW, Jüngel, A, Huscher, D, Huber, LC, Michel, BA, Gay, RE, Pisetsky, DS, Gay, S, Matucci-Cerinic, M, and Distler, O. "The relationship between plasma microparticles and disease manifestations in patients with systemic sclerosis." Arthritis Rheum 58.9 (September 2008): 2845-2853.
PMID
18759303
Source
pubmed
Published In
Arthritis and Rheumatism
Volume
58
Issue
9
Publish Date
2008
Start Page
2845
End Page
2853
DOI
10.1002/art.23735

Induction of plasma (TRAIL), TNFR-2, Fas ligand, and plasma microparticles after human immunodeficiency virus type 1 (HIV-1) transmission: implications for HIV-1 vaccine design.

The death of CD4(+) CCR5(+) T cells is a hallmark of human immunodeficiency virus (HIV) infection. We studied the plasma levels of cell death mediators and products--tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL), Fas ligand, TNF receptor type 2 (TNFR-2), and plasma microparticles--during the earliest stages of infection following HIV type 1 (HIV-1) transmission in plasma samples from U.S. plasma donors. Significant plasma TRAIL level elevations occurred a mean of 7.2 days before the peak of plasma viral load (VL), while TNFR-2, Fas ligand, and microparticle level elevations occurred concurrently with maximum VL. Microparticles had been previously shown to mediate immunosuppressive effects on T cells and macrophages. We found that T-cell apoptotic microparticles also potently suppressed in vitro immunoglobulin G (IgG) and IgA antibody production by memory B cells. Thus, release of TRAIL during the onset of plasma viremia (i.e., the eclipse phase) in HIV-1 transmission may initiate or amplify early HIV-1-induced cell death. The window of opportunity for a HIV-1 vaccine is from the time of HIV-1 transmission until establishment of the latently infected CD4(+) T cells. Release of products of cell death and subsequent immunosuppression following HIV-1 transmission could potentially narrow the window of opportunity during which a vaccine is able to extinguish HIV-1 infection and could place severe constraints on the amount of time available for the immune system to respond to the transmitted virus.

Authors
Gasper-Smith, N; Crossman, DM; Whitesides, JF; Mensali, N; Ottinger, JS; Plonk, SG; Moody, MA; Ferrari, G; Weinhold, KJ; Miller, SE; Reich, CF; Qin, L; Self, SG; Shaw, GM; Denny, TN; Jones, LE; Pisetsky, DS; Haynes, BF
MLA Citation
Gasper-Smith, N, Crossman, DM, Whitesides, JF, Mensali, N, Ottinger, JS, Plonk, SG, Moody, MA, Ferrari, G, Weinhold, KJ, Miller, SE, Reich, CF, Qin, L, Self, SG, Shaw, GM, Denny, TN, Jones, LE, Pisetsky, DS, and Haynes, BF. "Induction of plasma (TRAIL), TNFR-2, Fas ligand, and plasma microparticles after human immunodeficiency virus type 1 (HIV-1) transmission: implications for HIV-1 vaccine design." J Virol 82.15 (August 2008): 7700-7710.
PMID
18508902
Source
pubmed
Published In
Journal of virology
Volume
82
Issue
15
Publish Date
2008
Start Page
7700
End Page
7710
DOI
10.1128/JVI.00605-08

Expression of high mobility group protein 1 in the sera of patients and mice with systemic lupus erythematosus.

Authors
Jiang, W; Pisetsky, DS
MLA Citation
Jiang, W, and Pisetsky, DS. "Expression of high mobility group protein 1 in the sera of patients and mice with systemic lupus erythematosus." Ann Rheum Dis 67.5 (May 2008): 727-728. (Letter)
PMID
18408114
Source
pubmed
Published In
Annals of the rheumatic diseases
Volume
67
Issue
5
Publish Date
2008
Start Page
727
End Page
728
DOI
10.1136/ard.2007.074484

The induction of HMGB1 release from RAW 264.7 cells by transfected DNA.

High mobility group protein 1 (HMGB1) is a non-histone nuclear protein that can activate innate immunity when in an extracellular location. As shown in in vitro studies, while polyinosinic-polycytidylic acid [poly (I:C)] and LPS, TLR3 and TLR4 ligands, respectively, can induce HMGB1 release from macrophages, CpG DNA, a TLR 9 ligand, does not. Since DNA displays distinct immunostimulatory activity when transfected into cells, we investigated whether transfected DNA can induce HMGB1 release from macrophages. In these experiments, using RAW 264.7 cells as model, we show that DNA, either natural DNA or synthetic oligonucleotides, can induce HMGB1 release when used to stimulate cells with the transfection reagent Lipofectamine 2000; release occurred irrespective of the intrinsic activity of the DNA. The induction of HMGB1 release by transfected DNA was dependent on IFN-beta as shown by the inhibitory effects of an antibody. In addition, JNK activation mediated HMGB1 release induced by a transfected phosphorothioate oligonucleotide but not by transfected natural DNA. Together, these findings indicate that transfected DNA can stimulate macrophages to release HMGB1 under conditions in which free DNA is inactive and suggest a role of DNA in inducing inflammation when bound to molecules that influence its entry into cells.

Authors
Jiang, W; Pisetsky, DS
MLA Citation
Jiang, W, and Pisetsky, DS. "The induction of HMGB1 release from RAW 264.7 cells by transfected DNA." Mol Immunol 45.7 (April 2008): 2038-2044.
PMID
18031817
Source
pubmed
Published In
Molecular Immunology
Volume
45
Issue
7
Publish Date
2008
Start Page
2038
End Page
2044
DOI
10.1016/j.molimm.2007.10.019

Rheumatology in the current era: the challenge of success.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Rheumatology in the current era: the challenge of success." Nat Clin Pract Rheumatol 4.4 (April 2008): 165-.
PMID
18382480
Source
pubmed
Published In
Nature Clinical Practice Rheumatology
Volume
4
Issue
4
Publish Date
2008
Start Page
165
DOI
10.1038/ncprheum0791

The release of DNA into the plasma of mice following hepatic cell death by apoptosis and necrosis.

The goal of these investigations was to measure levels of DNA in the plasma of mice following administration of hepatotoxic agents to induce apoptotic or necrotic cell death and determine any differences in the release of this marker depending upon death pathway. For this purpose, the effects of varying doses of anti-Fas, acetaminophen (APAP) or carbon tetrachloride (CCl(4)) were assessed in normal mice. Plasma DNA was measured fluorometrically by the dye PicoGreen while lactate dehydrogenase (LDH) and caspase 3, other molecules released with cell injury or death, were measured by enzymatic assays. Histology was used to assess the occurrence of apoptosis or necrosis. Results of these experiments indicate that increased blood DNA levels occurred with all three agents and were highest with anti-Fas and CCl(4); caspase 3 levels were much higher with anti-Fas than the other agents. Histological examination confirmed the predominance of apoptotic death with anti-Fas and necrotic death with APAP and CCl(4). These results indicate that increased blood DNA is common in hepatotoxic injury and is a feature of both apoptotic and necrotic death.

Authors
Tran, TT; Groben, P; Pisetsky, DS
MLA Citation
Tran, TT, Groben, P, and Pisetsky, DS. "The release of DNA into the plasma of mice following hepatic cell death by apoptosis and necrosis." Biomarkers 13.2 (March 2008): 184-200.
PMID
18270870
Source
pubmed
Published In
Biomarkers (Informa)
Volume
13
Issue
2
Publish Date
2008
Start Page
184
End Page
200
DOI
10.1080/13547500701791719

A landmark study on treatment strategies for rheumatoid arthritis.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "A landmark study on treatment strategies for rheumatoid arthritis." Arthritis Rheum 58.2 Suppl (February 2008): S123-S125.
PMID
18240202
Source
pubmed
Published In
Arthritis and Rheumatism
Volume
58
Issue
2 Suppl
Publish Date
2008
Start Page
S123
End Page
S125
DOI
10.1002/art.23066

Pivotal advance: inhibition of HMGB1 nuclear translocation as a mechanism for the anti-rheumatic effects of gold sodium thiomalate.

Gold compounds such as gold sodium thiomalate (GST) can reduce the symptoms of rheumatoid arthritis (RA), although their mechanism of action is not well defined. As the proinflammatory mediator high mobility group box chromosomal protein 1 (HMGB1) may play a role in the pathogenesis of RA, we have performed in vitro studies to investigate whether GST inhibits HMGB1 release as the basis of its mode of action. Murine RAW 264.7 or human THP-1 macrophage cells were stimulated in culture with agents causing extracellular HMGB1 release, including LPS, IFN-gamma, polyinosinic:polycytidylic acid, IFN-beta, or NO in the presence of GST, ranging from 0 microM to 250 microM. Secretion and intracellular location of HMGB1 were assessed by Western blotting, HMGB1-specific ELISPOT assay, and immunofluorescent staining. In parallel, TNF and IFN-beta levels were analyzed by ELISPOT and/or ELISA. Supernatant NO production was analyzed by the Griess method. At pharmacologically relevant doses, GST inhibited the extracellular release of HMGB1 from activated macrophages and caused the nuclear retention of this protein; in contrast, no effects were observed on the secretion or production of TNF. Release of the key endogenous mediators of HMGB1 translocation, IFN-beta and NO, was inhibited by GST. This inhibition required gold, as sodium thiomalate did not affect the responses measured. Furthermore, gold chloride also inhibited release of HMGB1. Together, these results suggest a new mechanism for the anti-rheumatic effects of gold salts in RA and the potential of drugs, which interfere with intracellular HMGB1 transport mechanisms, as novel agents to treat RA.

Authors
Zetterström, CK; Jiang, W; Wähämaa, H; Ostberg, T; Aveberger, A-C; Schierbeck, H; Lotze, MT; Andersson, U; Pisetsky, DS; Erlandsson Harris, H
MLA Citation
Zetterström, CK, Jiang, W, Wähämaa, H, Ostberg, T, Aveberger, A-C, Schierbeck, H, Lotze, MT, Andersson, U, Pisetsky, DS, and Erlandsson Harris, H. "Pivotal advance: inhibition of HMGB1 nuclear translocation as a mechanism for the anti-rheumatic effects of gold sodium thiomalate." J Leukoc Biol 83.1 (January 2008): 31-38.
PMID
17913975
Source
pubmed
Published In
Journal of leukocyte biology
Volume
83
Issue
1
Publish Date
2008
Start Page
31
End Page
38
DOI
10.1189/jlb.0507323

The role of cell death in the pathogenesis of autoimmune disease: HMGB1 and microparticles as intercellular mediators of inflammation.

Cell death is critical to normal homeostasis, although this process, when increased aberrantly, can lead to the production of pro-inflammatory mediators promoting autoimmunity. Two novel intercellular mediators of inflammation generated during cell death are high mobility group box 1 (HMGB1) protein and microparticles (MPs). HMGB1 is a nuclear protein that functions in transcription when inside the nucleus but takes on pro-inflammatory properties when released during cell death. Microparticles are small, membrane-bound structures that extrude from cells when they die and contain cell surface proteins and nuclear material from their parent cells. MPs circulate widely throughout the vasculature and mediate long-distance communication between cells. Both MPs and HMGB1 have been implicated in the pathogenesis of a broad spectrum of inflammatory diseases, including the prototypic autoimmune conditions systemic lupus erythematosus and rheumatoid arthritis. Given their range of activity and association with active disease, both structures may prove to be targets for effective therapy in these and other disorders.

Authors
Ardoin, SP; Pisetsky, DS
MLA Citation
Ardoin, SP, and Pisetsky, DS. "The role of cell death in the pathogenesis of autoimmune disease: HMGB1 and microparticles as intercellular mediators of inflammation." Mod Rheumatol 18.4 (2008): 319-326. (Review)
PMID
18418695
Source
pubmed
Published In
Modern Rheumatology
Volume
18
Issue
4
Publish Date
2008
Start Page
319
End Page
326
DOI
10.1007/s10165-008-0054-z

Developments in the scientific understanding of lupus.

Systemic lupus erythematosus is a systemic autoimmune disease characterized by the production of antinuclear antibodies (ANAs). Recent research into human and murine lupus suggests that disease susceptibility results from genetic polymorphisms regulating immune responses as well as impairing the clearance of apoptotic cells. Because the products of dead cells, including nucleic acids, have immunologic activity, this situation can promote antigen-driven ANA responses. Furthermore, immune complexes of ANAs can drive the production of proinflammatory cytokines, inducing the 'interferon signature', and intensifying disease. Together, these findings point to new genetic and immunologic markers of disease as well as targets for new therapies.

Authors
Ardoin, SP; Pisetsky, DS
MLA Citation
Ardoin, SP, and Pisetsky, DS. "Developments in the scientific understanding of lupus." Arthritis Res Ther 10.5 (2008): 218-. (Review)
PMID
18947369
Source
pubmed
Published In
Arthritis Research and Therapy
Volume
10
Issue
5
Publish Date
2008
Start Page
218
DOI
10.1186/ar2488

High-mobility group box protein 1 (HMGB1): an alarmin mediating the pathogenesis of rheumatic disease.

High-mobility group box protein 1 (HMGB1) is a non-histone nuclear protein that has a dual function. Inside the cell, HMGB1 binds DNA, regulating transcription and determining chromosomal architecture. Outside the cell, HMGB1 can serve as an alarmin to activate the innate system and mediate a wide range of physiological and pathological responses. To function as an alarmin, HMGB1 translocates from the nucleus of the cell to the extra-cellular milieu, a process that can take place with cell activation as well as cell death. HMGB1 can interact with receptors that include RAGE (receptor for advanced glycation endproducts) as well as Toll-like receptor-2 (TLR-2) and TLR-4 and function in a synergistic fashion with other proinflammatory mediators to induce responses. As shown in studies on patients as well as animal models, HMGB1 can play an important role in the pathogenesis of rheumatic disease, including rheumatoid arthritis, systemic lupus erythematosus, and polymyositis among others. New approaches to therapy for these diseases may involve strategies to inhibit HMGB1 release from cells, its interaction with receptors, and downstream signaling.

Authors
Pisetsky, DS; Erlandsson-Harris, H; Andersson, U
MLA Citation
Pisetsky, DS, Erlandsson-Harris, H, and Andersson, U. "High-mobility group box protein 1 (HMGB1): an alarmin mediating the pathogenesis of rheumatic disease." Arthritis Res Ther 10.3 (2008): 209-. (Review)
PMID
18598385
Source
pubmed
Published In
Arthritis Research and Therapy
Volume
10
Issue
3
Publish Date
2008
Start Page
209
DOI
10.1186/ar2440

Microparticles stimulate the synthesis of prostaglandin E(2) via induction of cyclooxygenase 2 and microsomal prostaglandin E synthase 1.

OBJECTIVE: Microparticles are small vesicles that are released from activated or dying cells and that occur abundantly in the synovial fluid of patients with rheumatoid arthritis (RA). The goal of these studies was to elucidate the mechanisms by which microparticles activate synovial fibroblasts to express a proinflammatory phenotype. METHODS: Microparticles from monocytes and T cells were isolated by differential centrifugation. Synovial fibroblasts were cocultured with increasing numbers of microparticles. Gene expression was analyzed by real-time polymerase chain reaction and confirmed by Western blotting and enzyme immunoassay. Arachidonic acid labeled with tritium was used to study the transport of biologically active lipids by microparticles. The roles of NF-kappaB and activator protein 1 (AP-1) signaling were analyzed with electrophoretic mobility shift assay and transfection with small interfering RNA and IkappaB expression vectors. RESULTS: Microparticles strongly induced the synthesis of cyclooxygenase 2 (COX-2), microsomal prostaglandin E synthase 1 (mPGES-1), and prostaglandin E(2) (PGE(2)). In contrast, no up-regulation of COX-1, mPGES-2, cytosolic PGES, or phospholipase A(2) was observed. The induction of PGE(2) was blocked by selective inhibition of COX-2. Microparticles activated NF-kappaB, AP-1, p38, and JNK signaling in synovial fibroblasts. Inhibition of NF-kappaB, AP-1, and JNK signaling reduced the stimulatory effects. Arachidonic acid was transported from leukocytes to fibroblasts by microparticles. Arachidonic acid derived from microparticles was converted to PGE(2) by synovial fibroblasts. CONCLUSION: These results demonstrate that microparticles up-regulate the production of PGE(2) in synovial fibroblasts by inducing COX-2 and mPGES-1. These data provide evidence for a novel mechanism by which microparticles may contribute to inflammation and pain in RA.

Authors
Jüngel, A; Distler, O; Schulze-Horsel, U; Huber, LC; Ha, HR; Simmen, B; Kalden, JR; Pisetsky, DS; Gay, S; Distler, JHW
MLA Citation
Jüngel, A, Distler, O, Schulze-Horsel, U, Huber, LC, Ha, HR, Simmen, B, Kalden, JR, Pisetsky, DS, Gay, S, and Distler, JHW. "Microparticles stimulate the synthesis of prostaglandin E(2) via induction of cyclooxygenase 2 and microsomal prostaglandin E synthase 1." Arthritis Rheum 56.11 (November 2007): 3564-3574.
PMID
17968936
Source
pubmed
Published In
Arthritis and Rheumatism
Volume
56
Issue
11
Publish Date
2007
Start Page
3564
End Page
3574
DOI
10.1002/art.22980

Clinician's comment on the management of pain in arthritis.

The arthritic diseases are major sources of pain or disability, although they differ in etiology and treatment approach. For diseases such as RA, inflammation is the predominant mechanism that leads to systemic complaints such as pain as well as local destruction of cartilage and bone. In contrast, OA is primarily a degenerative process and, although inflammation may occur, it differs in quality and extent from that in the systemic inflammatory arthritidies. For both conditions, psychosocial interventions have significant positive benefits, but their application involves careful consideration of a variety of factors. These factors include the following: diagnosis, disease activity, damage, disease stage, patient age and demographics, presence of comorbidities, and availability of alternative or adjunctive approaches.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Clinician's comment on the management of pain in arthritis." Health Psychol 26.6 (November 2007): 657-659.
PMID
18020835
Source
pubmed
Published In
Health Psychology
Volume
26
Issue
6
Publish Date
2007
Start Page
657
End Page
659
DOI
10.1037/0278-6133.26.6.657

The role of microparticles in inflammation and thrombosis.

Microparticles (MP) are small membrane-bound vesicles that circulate in the peripheral blood and play active roles in thrombosis, inflammation and vascular reactivity. While MP can be released from nearly every cell type, most investigation has focused on MP of platelet, leucocyte and endothelial cell origin. Cells can release MP during activation or death. Flow cytometry is the usual method to quantify MP; the small size of these structures and lack of standardization in methodology complicate measurement. As MP contain surface and cytoplasmic contents of the parent cells and bear phosphatidylserine, antibodies to specific cell surface markers and annexin V can be used for identification. Through various mechanisms, MP participate in haemostasis and have procoagulant potential in disease. MP contribute to inflammation via their influence on cell-cell interactions and cytokine release, and MP also function in mediating vascular tone. In several disease states characterized by inflammation and vascular dysfunction, MP subpopulations are elevated, correlate with clinical events, and may have important roles in pathogenesis. In the rheumatic conditions such as rheumatoid arthritis and systemic lupus erythematosus, MP are potentially important markers of disease activity and have an increasingly recognized role in immunopathogenesis. It is clear that MP play an important role in atherosclerosis, and study of these structures may provide insight into the link between chronic inflammatory conditions and accelerated atherosclerosis. As biomarkers, MP allow access to usually inaccessible tissues such as the endothelium. Further research will hopefully lead to interventions targeting MP release and function.

Authors
Ardoin, SP; Shanahan, JC; Pisetsky, DS
MLA Citation
Ardoin, SP, Shanahan, JC, and Pisetsky, DS. "The role of microparticles in inflammation and thrombosis." Scand J Immunol 66.2-3 (August 2007): 159-165. (Review)
PMID
17635793
Source
pubmed
Published In
Scandinavian Journal of Immunology
Volume
66
Issue
2-3
Publish Date
2007
Start Page
159
End Page
165
DOI
10.1111/j.1365-3083.2007.01984.x

Role of Toll-like receptors in HMGB1 release from macrophages.

HMGB1 is a nonhistone nuclear protein that can serve as a cytokine and activate innate immunity. The translocation of this molecule from the inside to the outside of cells is a critical event in inflammation, occurring following activation of certain Toll-like receptors (TLRs) as well as during the course of apoptotic as well as necrotic cell death. Because the kinetics of HMGB1 release differs from that of a conventional cytokine, it provides a broader therapeutic window and may be an important new target of therapy for inflammatory, autoimmune, and infectious diseases.

Authors
Pisetsky, DS; Jiang, W
MLA Citation
Pisetsky, DS, and Jiang, W. "Role of Toll-like receptors in HMGB1 release from macrophages." Ann N Y Acad Sci 1109 (August 2007): 58-65.
PMID
17785291
Source
pubmed
Published In
Annals of the New York Academy of Sciences
Volume
1109
Publish Date
2007
Start Page
58
End Page
65
DOI
10.1196/annals.1398.008

The role of nuclear macromolecules in innate immunity.

Nuclear macromolecules, in addition to their intracellular role in regulating cell function, can translocate into the extracellular space where they can activate innate immunity. This translocation can occur in various settings and reflects the dynamic nature of nuclear structure. Of nuclear molecules, DNA and the DNA-binding protein, HMGB1, display distinct patterns of immune activity. For DNA, immune activity depends on sequence, base methylation, and context. While bacterial DNA is an immune activator, mammalian DNA is either inert or inhibitory when free. In contrast, mammalian DNA in the form of immune complexes can trigger immune cell activation. As shown in in vivo and in vitro studies, DNA can exit cells during apoptotic as well as necrotic cell death in a process that may depend on the presence of macrophages. Like DNA, HMGB1 can exit cells and acquire immune properties. For HMGB1, the translocation occurs in macrophages that have been stimulated by Toll-like receptor (TLR) ligands as well as cytokines; HMGB1 release can also occur with apoptotic as well as necrotic death. While HMGB1 alone can display cytokine activity, it may also activate cells in conjunction with other immune stimulators such as TLR ligands. For both DNA and HMGB1, the immune properties may therefore reflect the array of other endogenous as well as exogenous molecules present.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "The role of nuclear macromolecules in innate immunity." Proc Am Thorac Soc 4.3 (July 2007): 258-262. (Review)
PMID
17607009
Source
pubmed
Published In
Proceedings of the American Thoracic Society
Volume
4
Issue
3
Publish Date
2007
Start Page
258
End Page
262
DOI
10.1513/pats.200701-027AW

Autoimmunity: the nuclear arsenal of autoimmunity.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Autoimmunity: the nuclear arsenal of autoimmunity." Immunol Cell Biol 85.5 (July 2007): 344-345.
PMID
17533412
Source
pubmed
Published In
Immunology and cell biology
Volume
85
Issue
5
Publish Date
2007
Start Page
344
End Page
345
DOI
10.1038/sj.icb.7100074

The origin of extracellular DNA during the clearance of dead and dying cells.

DNA is a nuclear molecule that has both an intracellular and extracellular role. Inside the cell, it is the essential molecule of heredity while outside the cell it can have immunological activity, both alone and in the context of immune complexes. Furthermore, extracellular DNA has information content that can be mined by genomic techniques. Because of the association of extracellular DNA with clinical conditions marked by cell death, dead and dying cells have been considered the origin of this material. To investigate this process, in vitro and in vivo systems have been used to determine the release of DNA from cells, using Jurkat T cells as a model. Thus, in vitro, apoptotic Jurkat cells release DNA whereas necrotic cells do not. The presence of macrophages in these cultures, however, modifies the release process, causing release from necrotic cells as well. In in vivo experiments in which Jurkat cells are administered to normal mice, both apoptotic and necrotic cells give rise to DNA in the blood in a process that requires macrophages and can be modified by glucocorticoids. In this model, female and male mice differ in the extent of DNA release from the administered Jurkat cells. Together, these results indicate that, while apoptosis and necrosis can lead to a blood DNA response, this process requires macrophages and may be hormonally mediated.

Authors
Pisetsky, DS; Fairhurst, A-M
MLA Citation
Pisetsky, DS, and Fairhurst, A-M. "The origin of extracellular DNA during the clearance of dead and dying cells." Autoimmunity 40.4 (June 2007): 281-284. (Review)
PMID
17516210
Source
pubmed
Published In
Autoimmunity (Informa)
Volume
40
Issue
4
Publish Date
2007
Start Page
281
End Page
284
DOI
10.1080/08916930701358826

The relationship between apoptosis and high-mobility group protein 1 release from murine macrophages stimulated with lipopolysaccharide or polyinosinic-polycytidylic acid.

High-mobility group protein 1 (HMGB1) is a nonhistone nuclear protein whose function depends on cellular location. Inside the cell, HMGB1 modulates a variety of important cellular processes, including transcription, whereas outside the cell, HMGB1 acts as a cytokine that can promote inflammation and mediate sepsis and arthritis in animal models. In in vitro studies, proinflammatory molecules such as LPS, lipoteichoic acid, polyinosinic-polycytidylic acid (poly(I:C)), TNF-alpha, and type I and II IFNs can induce HMGB1 release from macrophages. Although these agents can activate cells, they can also induce apoptosis under certain circumstances. Therefore, because of evidence that apoptotic as well as necrotic cells can contribute to HMGB1-mediated events in sepsis, we have investigated the relationship between apoptosis and HMGB1 release in macrophages and other cells. In these experiments, using RAW 264.7 cells as a model, LPS and poly(I:C) caused HMGB1 release into the medium whereas CpG ODN failed to induce this response. With both LPS and poly(I:C), the extent of HMGB1 release correlated with the occurrence of apoptosis as measured by caspase 3 activation, lactate dehydrogenase release, and TUNEL staining. Similar results were obtained with primary murine macrophages as well as human Jurkat T cells. For Jurkat cells, poly(I:C) and NO donors induced apoptosis as well as HMGB1 release. Together, these results indicate that HMGB1 release from macrophages is correlated with the occurrence of apoptosis and suggest that these processes reflect common mechanisms and can occur concomitantly.

Authors
Jiang, W; Bell, CW; Pisetsky, DS
MLA Citation
Jiang, W, Bell, CW, and Pisetsky, DS. "The relationship between apoptosis and high-mobility group protein 1 release from murine macrophages stimulated with lipopolysaccharide or polyinosinic-polycytidylic acid." J Immunol 178.10 (May 15, 2007): 6495-6503.
PMID
17475879
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
178
Issue
10
Publish Date
2007
Start Page
6495
End Page
6503

Early rheumatoid arthritis.

PURPOSE OF REVIEW: Rheumatoid arthritis is a chronic inflammatory disease in which early aggressive therapy with disease-modifying antirheumatic drugs can improve outcome and prevent joint damage. While such therapy is effective, its application can be limited by diagnostic uncertainty in patients with early inflammatory arthritis and concerns about treatment of patients whose disease would remit spontaneously. The purpose of current research is therefore to identify prognostic markers of early disease and to determine the role of aggressive treatment strategies in inducing remission in such patients. RECENT FINDINGS: Recent research has provided new information on genetic markers predicting rapid progression of joint destruction; the role of serology, in particularly, antibodies to citrullinated peptides in diagnosing rheumatoid arthritis; the utility of radiographic techniques in detecting both early synovitis and bone erosion; and the value of combination therapy in controlling signs, symptoms and radiographic progression. Recent clinical studies support the efficacy of a combination of methotrexate with a biological agent, especially a tumor-necrosis-factor blocker, in reducing disease activity. SUMMARY: While current treatment approaches can produce significant benefits in patients with early arthritis, future investigation is needed to target therapy more selectively and to determine which patients respond best to various agents or combinations.

Authors
Mitchell, KL; Pisetsky, DS
MLA Citation
Mitchell, KL, and Pisetsky, DS. "Early rheumatoid arthritis." Curr Opin Rheumatol 19.3 (May 2007): 278-283. (Review)
PMID
17414956
Source
pubmed
Published In
Current Opinion in Rheumatology
Volume
19
Issue
3
Publish Date
2007
Start Page
278
End Page
283
DOI
10.1097/BOR.0b013e32805e87bf

Formative research in clinical trial development: attitudes of patients with arthritis in enhancing prevention trials.

In preparation for randomised controlled trials (RCTs) of disease-modifying antirheumatic drugs in patients with early inflammatory arthritis (EIA), formative research was conducted to enhance the design of such trials. The objectives of this research were to (1) determine patients' educational needs as they relate to the necessary elements of informed consent; and (2) assess patients' interest in enrolling in a hypothetical prevention trial. In-depth interviews were conducted with nine patients. Seven patients were women and all but one white. The mean age was 48 years. During the 4-month enrolment period, only three patients with EIA were identified; six patients with longer duration of symptoms were also interviewed. Most patients were able to express the primary aim of a hypothetical prevention trial presented. Factors cited by patients favouring enrolment were potential for direct medical benefit and knowledge that they would be withdrawn from the trial if they developed symptoms. Factors cited by patients against enrolment were the inclusion of a placebo and general uncertainty regarding treatment required by the RCT design. Pending larger-scale empirical projects to explore patients' attitudes about prevention trials, small-scale formative research in advance of such trials ought to be conducted.

Authors
Taylor, HA; Sugarman, J; Pisetsky, DS; Bathon, J
MLA Citation
Taylor, HA, Sugarman, J, Pisetsky, DS, and Bathon, J. "Formative research in clinical trial development: attitudes of patients with arthritis in enhancing prevention trials." Ann Rheum Dis 66.4 (April 2007): 542-544.
PMID
16984939
Source
pubmed
Published In
Annals of the rheumatic diseases
Volume
66
Issue
4
Publish Date
2007
Start Page
542
End Page
544
DOI
10.1136/ard.2006.059600

The role of membrane lipids in the induction of macrophage apoptosis by microparticles.

Microparticles are membrane-derived vesicles that are released from cells during activation or cell death. These particles can serve as mediators of intercellular cross-talk and induce a variety of cellular responses. Previous studies have shown that macrophages undergo apoptosis after phagocytosing microparticles. Here, we have addressed the hypothesis that microparticles trigger this process via lipid pathways. In these experiments, microparticles induced apoptosis in primary macrophage cells or cell lines (RAW 264.7 or U937) with up to a 5-fold increase. Preincubation of macrophages with phosphatidylinositol-3,5-bisphosphate (PtdIns(3,5)BP) reduced the microparticle-induced apoptosis in a dose-dependent manner. PtdIns(3,5)BP is a specific inhibitor of the acid sphingomyelinase and thus can block the generation of pro-apoptotic ceramides. Similarly, the pre-incubation of macrophages with PtdIns(3,5)BP prevented microparticle-induced upregulation of caspase 8, which is a major target molecule of ceramide action in the apoptosis pathway. PtdIns(3,5)BP, however, had no effect on the spontaneous rate of apoptosis. To evaluate further signaling pathways induced by microparticles, the extracellular signal regulated kinase (ERK-) 1 was investigated. This kinase plays a role in activating phospholipases A2 which cleaves membrane phospholipids into arachidonic acid; microparticles have been suggested to be a preferred substrate for phospholipases A2. As shown in our experiments, microparticles strongly increased the amount of phosphorylated ERK1/2 in RAW 264.7 macrophages in a time-dependent manner, peaking 15 min after co-incubation. Addition of PD98059, a specific inhibitor of ERK1, prevented the increase in apoptosis of RAW 264.7 macrophages. Together, these data suggest that microparticles perturb lipid homeostasis of macrophages and thereby induce apoptosis. These results emphasize the importance of biolipids in the cellular cross-talk of immune cells. Based on the fact that in clinical situations with excessive cell death such as malignancies, autoimmune diseases and following chemotherapies high levels of circulating microparticles might modulate phagocytosing cells, a suppression of the immune response might occur due to loss of macrophages.

Authors
Huber, LC; Jüngel, A; Distler, JHW; Moritz, F; Gay, RE; Michel, BA; Pisetsky, DS; Gay, S; Distler, O
MLA Citation
Huber, LC, Jüngel, A, Distler, JHW, Moritz, F, Gay, RE, Michel, BA, Pisetsky, DS, Gay, S, and Distler, O. "The role of membrane lipids in the induction of macrophage apoptosis by microparticles." Apoptosis 12.2 (February 2007): 363-374.
PMID
17191114
Source
pubmed
Published In
Apoptosis
Volume
12
Issue
2
Publish Date
2007
Start Page
363
End Page
374
DOI
10.1007/s10495-006-0622-7

Vaccination against the forkhead family transcription factor Foxp3 enhances tumor immunity.

Depletion of CD4+CD25+ regulatory T cells (Treg) by treatment with alphaCD25 antibody synergizes with vaccination protocols to engender protective immunity in mice. The effectiveness of targeting CD25 to eliminate Treg is limited by the fact that CD25, the low-affinity interleukin-2 receptor, is up-regulated on conventional T cells. At present, foxp3 is the only product known to be exclusively expressed in Treg of mice. However, foxp3 is not expressed on the cell surface and hence cannot be targeted with antibodies. In this study, we tested the hypothesis that vaccination of mice against foxp3, a self-antigen expressed also in the thymus, is capable of stimulating foxp3-specific CTL that will cause the depletion of Treg and enhanced antitumor immunity. Vaccination of mice with foxp3 mRNA-transfected dendritic cells elicited a robust foxp3-specific CTL response and potentiated vaccine-induced protective immunity comparably with that of alphaCD25 antibody administration. In contrast to alphaCD25 antibody treatment, repeated foxp3 vaccination did not interfere with vaccine-induced protective immunity. Importantly, foxp3 vaccination led to the preferential depletion of foxp3-expressing Treg in the tumor but not in the periphery, whereas alphaCD25 antibody treatment led to depletion of Treg in both the tumor and the periphery. Targeting foxp3 by vaccination offers a specific and simpler protocol for the prolonged control of Treg that may be associated with reduced risk of autoimmunity, introducing an approach whereby specific depletion of cells is not limited to targeting products expressed on the cell surface.

Authors
Nair, S; Boczkowski, D; Fassnacht, M; Pisetsky, D; Gilboa, E
MLA Citation
Nair, S, Boczkowski, D, Fassnacht, M, Pisetsky, D, and Gilboa, E. "Vaccination against the forkhead family transcription factor Foxp3 enhances tumor immunity." Cancer Res 67.1 (January 1, 2007): 371-380.
PMID
17210720
Source
pubmed
Published In
Cancer Research
Volume
67
Issue
1
Publish Date
2007
Start Page
371
End Page
380
DOI
10.1158/0008-5472.CAN-06-2903

Mechanisms of Disease: the role of high-mobility group protein 1 in the pathogenesis of inflammatory arthritis.

High-mobility group protein 1 (HMG1) is a nonhistone nuclear protein that is a prototype of a dual-function alarmin whose immune activity is dependent upon its cellular location. Inside the cell, HMG1 binds to DNA and has a role in transcriptional regulation. Outside the cell, HMG1 acts as a cytokine and has activities that resemble those of tumor necrosis factor. The cytokine activities of HMG1 become manifest when this protein translocates from the nucleus to the cytoplasm and, eventually, into the external milieu; this translocation occurs during cell activation and cell death. Given its cytokine activity, HMG1 has been implicated in the pathogenesis of a broad range of immune-mediated diseases including arthritis. The role for this protein in arthritis was established by observations of the expression of HMG1 in synovial tissue of patients with rheumatoid arthritis as well as in the joints of animals used to model arthritis. Furthermore, in the mouse model of collagen-induced arthritis, treatment with antibodies to HMG1 or to an inhibitory domain of HMG1 can attenuate joint inflammation and damage. These studies identify a novel pathway in the pathogenesis of inflammatory arthritis, as well as a new target for biologic therapy.

Authors
Jiang, W; Pisetsky, DS
MLA Citation
Jiang, W, and Pisetsky, DS. "Mechanisms of Disease: the role of high-mobility group protein 1 in the pathogenesis of inflammatory arthritis." Nat Clin Pract Rheumatol 3.1 (January 2007): 52-58. (Review)
PMID
17203009
Source
pubmed
Published In
Nature Clinical Practice Rheumatology
Volume
3
Issue
1
Publish Date
2007
Start Page
52
End Page
58
DOI
10.1038/ncprheum0379

Antibodies to DNA

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Antibodies to DNA." Systemic Lupus Erythematosus: A Companion to Rheumatology (2007): 225-232.
Source
scival
Published In
Systemic Lupus Erythematosus: A Companion to Rheumatology
Publish Date
2007
Start Page
225
End Page
232
DOI
10.1016/B978-0-323-04434-9.50027-0

Microparticles as mediators of cellular cross-talk in inflammatory disease.

Microparticles are a heterogeneous population of membrane-coated vesicles which can be released from virtually all cell types during activation or apoptosis. Release occurs from the cell surface in an exogenous budding process involving local rearrangement of the cytoskeleton. Given their origin, these particles can be identified by staining for cell surface markers and annexin V. As shown in in vitro studies, microparticles may represent a novel subcellular element for intercellular communication in inflammation. Thus, microparticles can transfer chemokine receptors and arachidonic acid between cells, activate complement, promote leukocyte rolling and stimulate the release of pro-inflammatory mediators. Under certain conditions, however, microparticles may also exert anti-inflammatory properties by inducing immune cell apoptosis and the production of anti-inflammatory mediators. Microparticles may play an important role in the pathogenesis of rheumatologic diseases as evidenced by their elevation in diseases such as systemic sclerosis (SSc), systemic vasculitis and antiphospholipid antibody syndrome and correlation with clinical events. A role in inflammatory arthritis is suggested by the finding that leukocyte-derived microparticles induce the production of matrix metalloproteinases and cytokines by synovial fibroblasts. Together, these findings point to novel signaling pathways of cellular cross-talk that may operate along the spectrum of soluble cytokines and mediators of direct cell-cell contact.

Authors
Distler, JHW; Huber, LC; Gay, S; Distler, O; Pisetsky, DS
MLA Citation
Distler, JHW, Huber, LC, Gay, S, Distler, O, and Pisetsky, DS. "Microparticles as mediators of cellular cross-talk in inflammatory disease." Autoimmunity 39.8 (December 2006): 683-690. (Review)
PMID
17178565
Source
pubmed
Published In
Autoimmunity (Informa)
Volume
39
Issue
8
Publish Date
2006
Start Page
683
End Page
690
DOI
10.1080/08916930601061538

The extracellular release of HMGB1 during apoptotic cell death.

High mobility group box 1 protein (HMGB1) is a non-histone nuclear protein with dual function. Inside the cell, HMGB1 binds DNA and regulates transcription, whereas outside the cell, it serves as a cytokine and mediates the late effects of LPS. The movement of HMGB1 into the extracellular space has been demonstrated for macrophages stimulated with LPS as well as cells undergoing necrosis but not apoptosis. The differential release of HMGB1 during death processes could reflect the structure of chromatin in these settings as well as the mechanisms for HMGB1 translocation. Since apoptotic cells can release some nuclear molecules such as DNA to which HMGB1 can bind, we therefore investigated whether HMGB1 release can occur during apoptosis as well as necrosis. For this purpose, Jurkat cells were treated with chemical inducers of apoptosis (staurosporine, etoposide, or camptothecin), and HMGB1 release into the medium was assessed by Western blotting. Results of these experiments indicate that HMGB1 appears in the media of apoptotic Jurkat cells in a time-dependent manner and that this release can be reduced by Z-VAD-fmk. Panc-1 and U937 cells treated with these agents showed similar release. In addition, HeLa cells induced to undergo apoptosis showed HMGB1 release. Furthermore, we showed using confocal microscopy that HMGB1 and DNA change their nuclear location in Jurkat cells undergoing apoptosis. Together, these studies indicate that HMGB1 release can occur during the course of apoptosis as well as necrosis and suggest that the release process may vary with cell type.

Authors
Bell, CW; Jiang, W; Reich, CF; Pisetsky, DS
MLA Citation
Bell, CW, Jiang, W, Reich, CF, and Pisetsky, DS. "The extracellular release of HMGB1 during apoptotic cell death." Am J Physiol Cell Physiol 291.6 (December 2006): C1318-C1325.
PMID
16855214
Source
pubmed
Published In
American journal of physiology. Cell physiology
Volume
291
Issue
6
Publish Date
2006
Start Page
C1318
End Page
C1325
DOI
10.1152/ajpcell.00616.2005

The role of IFN-alpha and nitric oxide in the release of HMGB1 by RAW 264.7 cells stimulated with polyinosinic-polycytidylic acid or lipopolysaccharide.

High mobility group protein 1 (HMGB1) is a nonhistone nuclear protein with a dual function. Inside the cell, HMGB1 binds to DNA and modulates a variety of processes, including transcription. Outside the cell, HMGB1 displays cytokine activity and can promote inflammation, serving as a mediator in models of shock and arthritis. In in vitro studies, proinflammatory molecules such as LPS, lipoteichoic acid, dsRNA, TNF-alpha, and IFN-gamma can induce HMGB1 release from macrophages. To define further the release process, we investigated the role of the downstream mediators, NO and IFN-alpha, in the release of HMGB1 from RAW 264.7 macrophage cells stimulated with LPS or polyinosinic-polycytidylic acid (poly(I:C)). In these experiments, 1400W, an inhibitor of NO production by the inducible NO synthase, reduced HMGB1 release stimulated by LPS, but not poly(I:C), whereas neutralizing IFN-alpha prevented HMGB1 release induced by poly(I:C), but not LPS. The addition of an NO donor and rIFN-alpha to RAW 264.7 cells caused HMGB1 release. Furthermore, inhibition of JNK activation attenuated HMGB1 release induced by either LPS or poly(I:C). Analysis of bone marrow-derived macrophages stimulated by LPS or poly(I:C) showed patterns of HMGB1 release similar to those of RAW 264.7 cells. Together, these experiments indicate that, although both LPS and poly(I:C) induce HMGB1 release from RAW 264.7 cells and murine macrophages, the response is differentially dependent on NO and IFN-alpha.

Authors
Jiang, W; Pisetsky, DS
MLA Citation
Jiang, W, and Pisetsky, DS. "The role of IFN-alpha and nitric oxide in the release of HMGB1 by RAW 264.7 cells stimulated with polyinosinic-polycytidylic acid or lipopolysaccharide." J Immunol 177.5 (September 1, 2006): 3337-3343.
PMID
16920974
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
177
Issue
5
Publish Date
2006
Start Page
3337
End Page
3343

The generation of extracellular DNA in SLE: the role of death and sex.

DNA is a large macromolecule that plays a central role in the pathogenesis of systemic lupus erythematosus (SLE), serving as a target antigen of autoantibodies as well as a major component of immune complexes. These complexes can both promote immune disturbances as well as deposit in the kidney to incite inflammation. While the origin of anti-DNA autoantibodies in SLE has received intense investigation, the mechanisms by which DNA exits cells to form immune complexes in the circulation is not well understood. To determine the origin of DNA circulating in the blood in SLE, our laboratory has been using a murine model system to track the in vivo fate of DNA from Jurkat T cells that have been made apoptotic or necrotic in vitro and then administered to mice. Results of these studies indicate that DNA from apoptotic and necrotic cells appears in the blood in a time- and dose-dependent manner. Irrespective of origin, this DNA has properties of nucleosomes as shown by its molecular weight. The process of release requires the presence of macrophages and can be modified by glucocorticoids as well as inflammation. In addition, sex may play a role in the generation of extracellular DNA from dead cells as male and female mice differ in their responses in this model. Together, these studies clarify the origin of extracellular DNA circulating in the blood in SLE and suggest steps in this process that can be interdicted by novel therapy.

Authors
Pisetsky, DS; Jiang, N
MLA Citation
Pisetsky, DS, and Jiang, N. "The generation of extracellular DNA in SLE: the role of death and sex." Scand J Immunol 64.3 (September 2006): 200-204.
PMID
16918687
Source
pubmed
Published In
Scandinavian Journal of Immunology
Volume
64
Issue
3
Publish Date
2006
Start Page
200
End Page
204
DOI
10.1111/j.1365-3083.2006.01822.x

Fulfilling Koch's postulates of autoimmunity: anti-NR2 antibodies in mice and men.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Fulfilling Koch's postulates of autoimmunity: anti-NR2 antibodies in mice and men." Arthritis Rheum 54.8 (August 2006): 2349-2352.
PMID
16868989
Source
pubmed
Published In
Arthritis and Rheumatism
Volume
54
Issue
8
Publish Date
2006
Start Page
2349
End Page
2352
DOI
10.1002/art.22029

The effect of UVB on lupus skin: new light on the role of apoptosis in the pathogenesis of autoimmunity.

Authors
Huber, LC; Gay, S; Distler, O; Pisetsky, DS
MLA Citation
Huber, LC, Gay, S, Distler, O, and Pisetsky, DS. "The effect of UVB on lupus skin: new light on the role of apoptosis in the pathogenesis of autoimmunity." Rheumatology (Oxford) 45.5 (May 2006): 500-501.
PMID
16467364
Source
pubmed
Published In
Rheumatology
Volume
45
Issue
5
Publish Date
2006
Start Page
500
End Page
501
DOI
10.1093/rheumatology/kel036

The binding of sera of patients with SLE to bacterial and mammalian DNA.

Systemic lupus erythematosus (SLE) is a prototypic autoimmune disease characterized by antibodies to DNA (anti-DNA). Although these antibodies have features of antigen drive, the source of this DNA is not defined. To assess the potential role of foreign and self-DNA as driving antigens, the specificity of SLE sera for bacterial and mammalian DNA was evaluated. Micrococcus lysodeikticus (MC) and calf thymus (CT) DNA were tested as antigens, with absorption on CT DNA columns used to identify antibodies to antigenic sites on the two DNA. Among 9 sets of longitudinal sera tested, all showed binding to both DNA, and none showed exclusive or predominant binding to CT DNA. With absorbed sera, antibodies could be distinguished in terms of cross-reactive or selective binding to the DNAs. These findings suggest that anti-DNA antibodies vary in specificity and are consistent with a role of both foreign and self-DNA in anti-DNA induction.

Authors
Hamilton, KJ; Schett, G; Reich, CF; Smolen, JS; Pisetsky, DS
MLA Citation
Hamilton, KJ, Schett, G, Reich, CF, Smolen, JS, and Pisetsky, DS. "The binding of sera of patients with SLE to bacterial and mammalian DNA." Clin Immunol 118.2-3 (February 2006): 209-218.
PMID
16298553
Source
pubmed
Published In
Clinical Immunology
Volume
118
Issue
2-3
Publish Date
2006
Start Page
209
End Page
218
DOI
10.1016/j.clim.2005.10.009

Serum, urinary, and salivary nitric oxide in rheumatoid arthritis: complexities of interpreting nitric oxide measures.

Nitric oxide (NO) may play important roles in rheumatoid arthritis (RA). RA is an inflammatory disease involving joints and other systems including salivary glands. To assess NO production in RA patients, we compared levels of serum, urine, and salivary nitrite and nitrate (NOx) in patients with RA and normal subjects, and we examined the relationships of these measures to disease activity. Serum, urine, and NOx levels as well as renal creatinine, NOx clearance and fractional excretion rates were compared in 25 RA patients and 20 age- and gender-matched healthy controls. Subjects were hospitalized for 3 days and placed on a NOx restricted diet. NOx was assayed using nitrate reductase and the Griess reagent. RA activity was assessed using standard clinical and laboratory measures. While consuming a restricted diet for 3 days to eliminate the effects of oral intake of NOx, 24 hour urinary NOx excretion decreased in both RA patients and healthy controls. Urine NOx levels at all time points were not significantly different between RA patients and normal subjects. Serum NOx levels also decreased during the 3 days of NOx restriction, but RA patients had higher serum NOx levels at all time points compared with the control group. Likewise, serum NOx/creatinine ratios were higher in RA patients than in controls. Although basal salivary flow rate and tear flow were lower in RA patients, salivary NOx levels did not differ between normal and RA subjects. While renal creatinine clearance was not different between the two groups, we found that RA patients had lower renal NOx clearance and lower renal NOx fractional excretion. After correction of p values for multiple comparisons, there were no significant relationships for the RA group between measures of disease activity and the urinary NOx, serum NOx, or urinary NOx clearance. Despite interest in the use of NO as a marker of disease activity, alterations in renal NOx clearance and fractional excretion in RA make it difficult to assess in vivo NO production even with strict dietary restriction of NOx intake.

Authors
Weinberg, JB; Lang, T; Wilkinson, WE; Pisetsky, DS; St Clair, EW
MLA Citation
Weinberg, JB, Lang, T, Wilkinson, WE, Pisetsky, DS, and St Clair, EW. "Serum, urinary, and salivary nitric oxide in rheumatoid arthritis: complexities of interpreting nitric oxide measures." Arthritis Res Ther 8.5 (2006): R140-.
PMID
16907988
Source
pubmed
Published In
Arthritis Research and Therapy
Volume
8
Issue
5
Publish Date
2006
Start Page
R140
DOI
10.1186/ar2030

In vitro assay of immunostimulatory activities of plasmid vectors.

DNA vaccination represents a novel and potentially important approach to induce immune responses against protein antigens. In this approach, the vaccine is a plasmid DNA vector that can be taken up by cells to produce a protein, encoded by the vector, to be targeted for the induction of humoral or cellular responses. Although the intracellular production of the antigen may promote responses, the vectors themselves may display adjuvant activity because of their intrinsic immunostimulatory properties. These properties reflect sequence motifs, centering on an unmethylated CpG dinucleotide, which can trigger the TLR9 pattern recognition receptor. As shown by studies in vitro, plasmid DNA can stimulate B cells, macrophages, and dendritic cells, and trigger a broad range of pro-inflammatory responses. Because this stimulation results from common sequence motifs, the activity of a plasmid vector can be assessed by the in vitro assay of a limited number of responses, including proliferation of B cells as well as production of cytokines by macrophages or dendritic cells.

Authors
Jiang, W; Reich, CF; Pisetsky, DS
MLA Citation
Jiang, W, Reich, CF, and Pisetsky, DS. "In vitro assay of immunostimulatory activities of plasmid vectors." Methods Mol Med 127 (2006): 55-70. (Review)
PMID
16988446
Source
pubmed
Published In
Methods in Molecular Medicine
Volume
127
Publish Date
2006
Start Page
55
End Page
70
DOI
10.1385/1-59745-168-1:55

Rheumatology in 2006: crossroads or crisis?

Rheumatology has made remarkable advances in patient treatment in the past decade related to the impressive array of new drugs that have been approved or are undergoing clinical trial. While this situation should engender optimism for the future, concerns about sustaining momentum have been raised. These concerns relate to uncertainty in the research agenda for major diseases such as osteoarthritis and fibromyalgia, lack of informatics systems to allow accurate assessment of risks and benefits of new treatments, and a paucity of clinical trials in rheumatoid arthritis aimed at sustained remission or cure. Fortunately, the opportunities for the future remain very bright because of burgeoning research in biomedicine and outcomes assessment as well as progress in developing personalized medicine to individualize treatment better.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Rheumatology in 2006: crossroads or crisis?." Bull NYU Hosp Jt Dis 64.1-2 (2006): 9-11. (Review)
PMID
17121482
Source
pubmed
Published In
Bulletin of the NYU hospital for joint diseases
Volume
64
Issue
1-2
Publish Date
2006
Start Page
9
End Page
11

ICF core sets: how to specify impairment and function in systemic lupus erythematosus.

The World Health Organization's International Classification of Function (ICF) is a tool to characterize and illuminate better the full of array of problems a patient faces when affected by disease. Specifying these problems is a particular challenge in a disease like systemic lupus erythematosus (SLE) because of the wide variety in organ systems involved, its variable activity and severity, and considerable ethnic and local differences. The authors of this manuscript believe, however, that a broader understanding will prove essential for optimal patient care, and that there is sufficient experience now in defining ICF Core Sets to successfully complete core sets for SLE. Therefore, we will embark on an international project for developing ICF Core Sets for SLE, which we here delineate. This development will include two versions: 1) The Brief ICF Core Set for SLE will be a very focused list of categories essential for SLE clinical trials; and 2) The Comprehensive ICF Core Set will be much broader and useful for guiding multidisciplinary assessment in patients with SLE. Both Core Sets will be developed in a formal decision-making and consensus process of health professionals integrating evidence gathered from preliminary studies. The final definition of the Core Sets will occur at a consensus conference which will integrate: i) a systematic review of the literature regarding the outcome measures used in clinical trials and selected observational studies; ii) focus groups or semi-structured interviews with SLE patients; iii) a Delphi exercise with world wide involvement of experts; and iv) the evidence from empirical studies. The development of these SLE ICF Core Sets is designed to be an inclusive, open, worldwide process. We therefore invite both SLE clinical experts and SLE patients to participate actively.

Authors
Aringer, M; Stamm, TA; Pisetsky, DS; Yarboro, CH; Cieza, A; Smolen, JS; Stucki, G
MLA Citation
Aringer, M, Stamm, TA, Pisetsky, DS, Yarboro, CH, Cieza, A, Smolen, JS, and Stucki, G. "ICF core sets: how to specify impairment and function in systemic lupus erythematosus." Lupus 15.4 (2006): 248-253.
PMID
16686267
Source
pubmed
Published In
Lupus
Volume
15
Issue
4
Publish Date
2006
Start Page
248
End Page
253
DOI
10.1191/0961203306lu2298xx

Microparticles as regulators of inflammation: novel players of cellular crosstalk in the rheumatic diseases.

Authors
Distler, JHW; Pisetsky, DS; Huber, LC; Kalden, JR; Gay, S; Distler, O
MLA Citation
Distler, JHW, Pisetsky, DS, Huber, LC, Kalden, JR, Gay, S, and Distler, O. "Microparticles as regulators of inflammation: novel players of cellular crosstalk in the rheumatic diseases." Arthritis Rheum 52.11 (November 2005): 3337-3348. (Review)
PMID
16255015
Source
pubmed
Published In
Arthritis and Rheumatism
Volume
52
Issue
11
Publish Date
2005
Start Page
3337
End Page
3348
DOI
10.1002/art.21350

The effects of CpG DNA on HMGB1 release by murine macrophage cell lines.

DNA containing cytosine-guanine dinucleotide (CpG) motifs (CpG DNA) has potent immunostimulatory activities that resemble those of lipopolysaccharide (LPS) in its effects on the innate immune system. Among its activities, LPS can induce the release of high mobility group protein (HMGB1) by macrophages, a dual function molecule that can mediate the late effects of LPS. To determine whether CpG DNA can also induce HMGB1 release, the effects of a synthetic CpG oligonucleotide (ODN) on HMGB1 release from RAW 264.7 and J774A.1 cells were assessed by Western blotting of culture supernatants. Under conditions in which the CpG ODN activated the cell lines, as assessed by stimulation of tumor necrosis factor alpha and interleukin-12, it failed to cause HMGB1 release into the media. Although unable to induce HMGB1 release by itself, the CpG ODN nevertheless potentiated the action of LPS. With RAW 264.7 cells, lipoteichoic acid and polyinosinic-polycytidylic acid, like LPS, stimulated HMGB1 release as well as cytokine production. These results indicate that the effects of CpG DNA on macrophages differ from other ligands of Toll-like receptors and may lead to a distinct pattern of immune cell activation in the context of infection or its use as an immunomodulatory agent.

Authors
Jiang, W; Li, J; Gallowitsch-Puerta, M; Tracey, KJ; Pisetsky, DS
MLA Citation
Jiang, W, Li, J, Gallowitsch-Puerta, M, Tracey, KJ, and Pisetsky, DS. "The effects of CpG DNA on HMGB1 release by murine macrophage cell lines." J Leukoc Biol 78.4 (October 2005): 930-936.
PMID
16081598
Source
pubmed
Published In
Journal of leukocyte biology
Volume
78
Issue
4
Publish Date
2005
Start Page
930
End Page
936
DOI
10.1189/jlb.0405208

The influence of oxygen tension on the induction of nitric oxide and prostaglandin E2 by mechanical stress in articular cartilage.

OBJECTIVES: Articular cartilage is an avascular tissue that exists at low oxygen tension. Oxygen tension can influence the production of the pro-inflammatory mediators nitric oxide (NO) and prostaglandin E2 (PGE(2)) in cartilage, which are increased in osteoarthritis (OA). The synthesis of these molecules can be stimulated by mechanical stress, which is an important risk factor for OA. The objective of this study was to determine the influence of oxygen tension on the induction of NO and PGE(2) production in articular cartilage in response to mechanical stress. DESIGN: Intermittent mechanical compression (0.05MPa, 0.5Hz for 24h) was applied to full thickness skeletally mature porcine articular cartilage explants at either 20%, 5%, or 1% O(2). NO, PGE(2) and peroxynitrite formation were measured, and the effect of the selective nitric oxide synthase 2 inhibitor 1400W was tested. RESULTS: Incubating articular cartilage at 5% O(2) significantly increased (P<0.001) baseline NO production, as compared with 1% or 20% O(2). Peroxynitrite formation was lower at reduced oxygen tension. Mechanical compression significantly increased (P<0.001) NO production at 20% O(2) but not at 5% or 1% O(2), and significantly increased (P<0.001) PGE(2) production at 20% O(2) (50 fold) and 5% O(2) (4 fold) but not at 1% O(2). 1400W blocked mechanically induced NO production and further increased PGE(2) production at 5% O(2) (P<0.05). CONCLUSIONS: Oxygen tension influences the endogenous production of NO and PGE(2) in cartilage and can have a significant effect on the induction of these inflammatory mediators in response to mechanical compression.

Authors
Fermor, B; Weinberg, JB; Pisetsky, DS; Guilak, F
MLA Citation
Fermor, B, Weinberg, JB, Pisetsky, DS, and Guilak, F. "The influence of oxygen tension on the induction of nitric oxide and prostaglandin E2 by mechanical stress in articular cartilage." Osteoarthritis Cartilage 13.10 (October 2005): 935-941.
PMID
15975834
Source
pubmed
Published In
Osteoarthritis and Cartilage
Volume
13
Issue
10
Publish Date
2005
Start Page
935
End Page
941
DOI
10.1016/j.joca.2005.05.001

The release of microparticles by apoptotic cells and their effects on macrophages.

Microparticles are small membrane vesicles released from the cell membrane by exogenous budding. To elucidate the interactions of microparticles with macrophages, the effect of microparticles released from Jurkat T cells on RAW 264.7 cells was determined. Microparticles were isolated by differential centrifugation, using FACS analysis with annexin V and cell surface markers for identification. Various inducers of apoptosis increased the release of microparticles from Jurkat cells up to 5-fold. The released microparticles were then cultured with RAW 264.7 cells. As shown by confocal microscopy and FACS analysis, RAW 264.7 macrophages cleared microparticles by phagocytosis. In addition, microparticles induced apoptosis in RAW 264.7 cells in a dose-dependent manner with up to a 5-fold increase of annexin V positive cells and 9-fold increase in caspase 3 activity. Cell proliferation as determined by the MTT test was also reduced. Furthermore, microparticles stimulated the release of microparticles from macrophages. These effects were specific for macrophages, since no apoptosis was observed in NIH 3T3 and L929 cells. These findings indicate that microparticles can induce macrophages to undergo apoptosis, in turn resulting in a further increase of microparticles. The release of microparticles from apoptotic cells may therefore represent a novel amplification loop of cell death.

Authors
Distler, JHW; Huber, LC; Hueber, AJ; Reich, CF; Gay, S; Distler, O; Pisetsky, DS
MLA Citation
Distler, JHW, Huber, LC, Hueber, AJ, Reich, CF, Gay, S, Distler, O, and Pisetsky, DS. "The release of microparticles by apoptotic cells and their effects on macrophages." Apoptosis 10.4 (August 2005): 731-741.
PMID
16133865
Source
pubmed
Published In
Apoptosis
Volume
10
Issue
4
Publish Date
2005
Start Page
731
End Page
741
DOI
10.1007/s10495-005-2941-5

Role of thromboxane A2 in the induction of apoptosis of immature thymocytes by lipopolysaccharide.

Lipopolysaccharide (LPS) causes apoptotic deletion of CD4(+) CD8(+) thymocytes, a phenomenon that has been linked to immune dysfunction and poor survival during sepsis. Given the abundance of thromboxane-prostanoid (TP) receptors in CD4(+) CD8(+) thymocytes and in vitro evidence that thromboxane A(2) (TXA(2)) causes apoptosis of these cells, we tested whether enhanced generation of TXA(2) plays a role in LPS-induced thymocyte apoptosis. Mice injected with 50 micro LPS intraperitoneally displayed a marked increase in generation of TXA(2) and prostaglandin E(2) in the thymus as well as apoptotic deletion of CD4(+) CD8(+) thymocytes. Administration of indomethacin or rofecoxib inhibited prostanoid synthesis but did not affect thymocyte death. In contrast, thymocyte apoptosis in response to LPS was significantly attenuated in TP-deficient mice. These studies indicate that TXA(2) mediates a portion of apoptotic thymocyte death caused by LPS. The absence of an effect of global inhibition of prostanoid synthesis suggests a complex role for prostanoids in this model.

Authors
Rocha, PN; Plumb, TJ; Robinson, LA; Spurney, R; Pisetsky, D; Koller, BH; Coffman, TM
MLA Citation
Rocha, PN, Plumb, TJ, Robinson, LA, Spurney, R, Pisetsky, D, Koller, BH, and Coffman, TM. "Role of thromboxane A2 in the induction of apoptosis of immature thymocytes by lipopolysaccharide." Clin Diagn Lab Immunol 12.8 (August 2005): 896-903.
PMID
16085905
Source
pubmed
Published In
Clinical and diagnostic laboratory immunology
Volume
12
Issue
8
Publish Date
2005
Start Page
896
End Page
903
DOI
10.1128/CDLI.12.8.896-903.2005

The role of macrophages in the in vitro generation of extracellular DNA from apoptotic and necrotic cells.

Cell death is a ubiquitous process that occurs by apoptosis or necrosis depending on the triggering event. While apoptotic and necrotic cells differ biochemically, both are cleared by macrophages for elimination. The process is very efficient, although DNA can appear in the blood in various clinical conditions associated with cell death. To define the role of macrophages in the generation of extracellular DNA, in vitro experiments were performed, assessing the release of DNA into the media of apoptotic or necrotic Jurkat cells cultured with RAW264.7 or J774 macrophage cell lines. DNA was measured by a fluorimetric assay using the dye PicoGreen. In these experiments, while necrotic cells alone did not release DNA, in the presence of macrophages, significant amounts of DNA appeared in the medium. This DNA contained sequences from the Jurkat cells and had reduced molecular weight in comparison to cellular DNA. Furthermore, coculture of macrophages with enucleated necrotic Jurkat cells did not release DNA, suggesting that the DNA came from the dead cell. In contrast, Jurkat cells made apoptotic by treatment with either staurosporine or etoposide spontaneously released DNA, while coculture with macrophages caused a decrease in the DNA released. With apoptotic cells, the DNA in the medium showed low molecular weight and laddering whether or not macrophages were present. Together, these results indicate that macrophages play an important role in the generation of extracellular DNA from dead and dying cells, with the effect dependent on how the cell died.

Authors
Choi, J-J; Reich, CF; Pisetsky, DS
MLA Citation
Choi, J-J, Reich, CF, and Pisetsky, DS. "The role of macrophages in the in vitro generation of extracellular DNA from apoptotic and necrotic cells." Immunology 115.1 (May 2005): 55-62.
PMID
15819697
Source
pubmed
Published In
Immunology
Volume
115
Issue
1
Publish Date
2005
Start Page
55
End Page
62
DOI
10.1111/j.1365-2567.2005.02130.x

Toward optimal health: William P. Docken, M.D. and David S. Pisetsky, M.D., Ph.D. discuss arthritis in women. Interview by Jody R. Godfrey.

Authors
Docken, WP; Pisetsky, DS
MLA Citation
Docken, WP, and Pisetsky, DS. "Toward optimal health: William P. Docken, M.D. and David S. Pisetsky, M.D., Ph.D. discuss arthritis in women. Interview by Jody R. Godfrey." J Womens Health (Larchmt) 14.3 (April 2005): 208-213. (Interview)
PMID
15857266
Source
pubmed
Published In
Journal of Women's Health
Volume
14
Issue
3
Publish Date
2005
Start Page
208
End Page
213
DOI
10.1089/jwh.2005.14.208

The effect of inflammation on the generation of plasma DNA from dead and dying cells in the peritoneum.

To assess the effects of inflammation on the generation of circulating DNA from dead and dying cells, plasma DNA levels were determined in BALB/c mice, administered apoptotic or necrotic Jurkat cells following induction of peritonitis by treatment with thioglycollate (TG), peptone (PT), or sodium periodate (NaIO(4)). In mice receiving TG or NaIO(4), plasma DNA levels following intraperitoneal administration of Jurkat cells were significantly reduced compared with controls, whereas they were not affected in mice receiving PT. To determine the basis of these differences, the cellular composition of peritoneal fluids prior to the administration of the dead cells was analyzed. Among agents tested, TG administration led to the largest increase in cells, both neutrophils and monocytes. As shown by flow cytometry, the exudates contained apoptotic neutrophils and macrophages, with the highest levels in the TG-induced exudates. Analysis of DNA and caspase 3 in the fluids also showed differences. TG exudates showed increases in DNA and caspase 3, while NaIO(4)-induced exudates had an increase only in DNA. Fluid from PT-treated mice did not have increases in DNA or caspase 3. Together, these results indicate that prior inflammation can affect the generation of blood DNA from apoptotic or necrotic cells, although this effect may vary depending on the composition of the exudates with respect to cells as well as DNA.

Authors
Jiang, N; Pisetsky, DS
MLA Citation
Jiang, N, and Pisetsky, DS. "The effect of inflammation on the generation of plasma DNA from dead and dying cells in the peritoneum." J Leukoc Biol 77.3 (March 2005): 296-302.
PMID
15601668
Source
pubmed
Published In
Journal of leukocyte biology
Volume
77
Issue
3
Publish Date
2005
Start Page
296
End Page
302
DOI
10.1189/jlb.0704411

The induction of matrix metalloproteinase and cytokine expression in synovial fibroblasts stimulated with immune cell microparticles.

Rheumatoid arthritis is a chronic inflammatory disease characterized by destruction of cartilage and bone that is mediated by synovial fibroblasts. To determine the mechanisms by which these cells are activated to produce matrix metalloproteinases (MMPs), the effects of microparticles were investigated. Microparticles are small membrane-bound vesicles whose release from immune cells is increased during activation and apoptosis. Because microparticles occur abundantly in the synovial fluid in rheumatoid arthritis, they could represent novel stimulatory agents. Microparticles derived from T cells and monocytes strongly induced the synthesis of MMP-1, MMP-3, MMP-9, and MMP-13 in fibroblasts. The induction was time-dependent, with effects primarily observed after 36 h; under these conditions, MMP-2, MMP-14, and tissue inhibitor of MMP-1 (TIMP-1), TIMP-2, and TIMP-3 were not induced. Microparticles also increased the synthesis of inflammatory mediators including IL-6, IL-8, monocyte chemoattractant protein 1 (MCP-1), and MCP-2. In Ikappa-B-transfected synovial fibroblasts, MMPs were less inducible by microparticles compared with wild-type fibroblasts. Blocking of TNFalpha and IL-1beta with antibodies against TNFalpha and with IL-1 receptor antagonist did not abrogate stimulation by microparticles. These data provide evidence for a novel mechanism by which vesicles derived from activated or apoptotic immune cells can promote the destructive activity of synovial fibroblasts in rheumatoid arthritis.

Authors
Distler, JHW; Jüngel, A; Huber, LC; Seemayer, CA; Reich, CF; Gay, RE; Michel, BA; Fontana, A; Gay, S; Pisetsky, DS; Distler, O
MLA Citation
Distler, JHW, Jüngel, A, Huber, LC, Seemayer, CA, Reich, CF, Gay, RE, Michel, BA, Fontana, A, Gay, S, Pisetsky, DS, and Distler, O. "The induction of matrix metalloproteinase and cytokine expression in synovial fibroblasts stimulated with immune cell microparticles." Proc Natl Acad Sci U S A 102.8 (February 22, 2005): 2892-2897.
PMID
15701693
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
102
Issue
8
Publish Date
2005
Start Page
2892
End Page
2897
DOI
10.1073/pnas.0409781102

Detailing ethnicity and phenotypes is critical for pooling association studies: Comment on the article by Huizinga et al. [5] (multiple letters)

Authors
Rahman, P; Huizinga, TWJ; Pisetsky, DS; Kimberly, RP
MLA Citation
Rahman, P, Huizinga, TWJ, Pisetsky, DS, and Kimberly, RP. "Detailing ethnicity and phenotypes is critical for pooling association studies: Comment on the article by Huizinga et al. [5] (multiple letters)." Arthritis and Rheumatism 52.2 (2005): 676--.
PMID
15692998
Source
scival
Published In
Arthritis and Rheumatism
Volume
52
Issue
2
Publish Date
2005
Start Page
676-
DOI
10.1002/art.20814

The effects of cyclic mechanical strain and tumor necrosis factor alpha on the response of cells of the meniscus.

OBJECTIVES: Cells of the knee meniscus respond to changes in their biochemical and biomechanical environments with alterations in the biosynthesis of matrix constituents and inflammatory mediators. Tumor necrosis factor alpha (TNF-alpha) is a pro-inflammatory cytokine that is involved in the pathogenesis of both osteoarthritis and rheumatoid arthritis, but its influence on meniscal physiology or mechanobiology is not fully understood. The objectives of this study were to examine the hypothesis that cyclic mechanical strain of meniscal cells modulates the biosynthesis of matrix macromolecules and pro-inflammatory mediators, and to determine if this response is altered by TNF-alpha. METHODS: Cells were isolated from the inner two-thirds of porcine medial menisci and subjected to biaxial tensile strain of 5-15% at a frequency of 0.5Hz. The synthesis of proteoglycan, protein, nitric oxide (NO), and prostaglandin E(2) were determined. RESULTS: Cyclic tensile strain increased the production of nitric oxide through the upregulation of nitric oxide synthase 2 (NOS2) and also increased synthesis rates of prostaglandin E(2), proteoglycan, and total protein in a manner that depended on strain magnitude. TNF-alpha increased the production of NO and total protein, but inhibited proteoglycan synthesis rates. TNF-alpha prevented the mechanical stimulation of proteoglycan synthesis, and this effect was not dependent on NOS2. CONCLUSIONS: These findings indicate that pro-inflammatory cytokines can modulate the responses of meniscal cells to mechanical signals, suggesting that both biomechanical and inflammatory factors could contribute to the progression of joint disease as a consequence of altered loading of the meniscus.

Authors
Fermor, B; Jeffcoat, D; Hennerbichler, A; Pisetsky, DS; Weinberg, JB; Guilak, F
MLA Citation
Fermor, B, Jeffcoat, D, Hennerbichler, A, Pisetsky, DS, Weinberg, JB, and Guilak, F. "The effects of cyclic mechanical strain and tumor necrosis factor alpha on the response of cells of the meniscus." Osteoarthritis Cartilage 12.12 (December 2004): 956-962.
PMID
15564062
Source
pubmed
Published In
Osteoarthritis and Cartilage
Volume
12
Issue
12
Publish Date
2004
Start Page
956
End Page
962
DOI
10.1016/j.joca.2004.08.007

The immune response to cell death in SLE.

Systemic lupus erythematosus (SLE) is a prototypic autoimmune disease characterized by the production of antinuclear antibodies (ANA). These antibodies target a wide variety of antigens whose presence in an immunologically active form may result from cell death processes that cause their translocation and release from cells. As indicated by in vivo model systems, the release of DNA from cells may not be a simple consequence of cell death but rather may require the intervention of other cell types including macrophages. Thus, in mice, administration of either apoptotic or necrotic cells produces a blood DNA response, whereas mice lacking macrophages fail to show blood DNA under the same conditions. Furthermore, the circulating DNA arising from apoptotic and necrotic cells displays a similar pattern with respect to size distribution, with both showing DNA laddering, a pattern indicating enzymatic cleavage. Since circulating DNA in the form of immune complexes can play a role in lupus pathogenesis, these findings suggest that the generation and clearance of dead cells are important events that may underlie autoimmunity in this disease and may be targeted for therapy.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "The immune response to cell death in SLE." Autoimmun Rev 3.7-8 (November 2004): 500-504.
PMID
15546797
Source
pubmed
Published In
Autoimmunity Reviews
Volume
3
Issue
7-8
Publish Date
2004
Start Page
500
End Page
504
DOI
10.1016/j.autrev.2004.07.010

Toll-like receptors in the pathogenesis of human disease.

Members of the Toll-like receptor (TLR) family are key regulators of both innate and adaptive immune responses. The function of TLRs in various human diseases has been investigated by comparison of the incidence of disease among people having different polymorphisms in genes that participate in TLR signaling. These studies have shown that TLR function affects several diseases, including sepsis, immunodeficiencies, atherosclerosis and asthma. As this body of data grows, it will provide new insights into disease pathogenesis as well as valuable information on the merits of various therapeutic options.

Authors
Cook, DN; Pisetsky, DS; Schwartz, DA
MLA Citation
Cook, DN, Pisetsky, DS, and Schwartz, DA. "Toll-like receptors in the pathogenesis of human disease." Nat Immunol 5.10 (October 2004): 975-979. (Review)
PMID
15454920
Source
pubmed
Published In
Nature Immunology
Volume
5
Issue
10
Publish Date
2004
Start Page
975
End Page
979
DOI
10.1038/ni1116

DNA as a marker of cell death in systemic lupus erythematosus.

DNA circulates in the blood in systemic lupus erythematosus, among other conditions, and plays a role in immunopathogenesis in the form of immune complexes. As shown in experiments in mice, blood DNA levels rise following treatments to induce apoptosis and the administration of cells made apoptotic or necrotic in vitro. In mice lacking macrophage function, however, blood levels do not rise following administration of dead cells. These results indicate that circulating DNA may be a marker of cell death, although its levels likely reflect a complex process involving the interactions of macrophages with dead and dying cells.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "DNA as a marker of cell death in systemic lupus erythematosus." Rheum Dis Clin North Am 30.3 (August 2004): 575-x. (Review)
PMID
15261342
Source
pubmed
Published In
Rheumatic Disease Clinics of North America
Volume
30
Issue
3
Publish Date
2004
Start Page
575
End Page
x
DOI
10.1016/j.rdc.2004.04.009

Associations, populations, and the truth: recommendations for genetic association studies in Arthritis & Rheumatism.

Authors
Huizinga, TWJ; Pisetsky, DS; Kimberly, RP
MLA Citation
Huizinga, TWJ, Pisetsky, DS, and Kimberly, RP. "Associations, populations, and the truth: recommendations for genetic association studies in Arthritis & Rheumatism." Arthritis Rheum 50.7 (July 2004): 2066-2071. (Review)
PMID
15248203
Source
pubmed
Published In
Arthritis and Rheumatism
Volume
50
Issue
7
Publish Date
2004
Start Page
2066
End Page
2071
DOI
10.1002/art.20360

Release of DNA from dead and dying lymphocyte and monocyte cell lines in vitro.

DNA is a nuclear macromolecule that circulates in the blood where its levels can reflect the activity of inflammatory and malignant diseases. While dead and dying cells have usually been considered the source of blood DNA, the mechanisms for its release during apoptosis and necrosis are not well defined. To elucidate DNA release, an in vitro model system was used, assessing DNA in the media of living, apoptotic or necrotic Jurkat and U937 cells. Apoptosis was induced by etoposide, camptothecin or staurosporine, while necrosis was induced by heating at 56 degrees C. DNA release was measured by fluorometry with the dye PicoGreen while the extent of death was measured by fluorescence-activated cell sorter analysis with propidium iodide and annexin. Apoptotic Jurkat cells released significantly more DNA in the media than untreated cells while necrotic cells did not show significant DNA release. U937 cells showed similar findings. Pretreatment of Jurkat cells with z-VAD-fmk, a caspase inhibitor, reduced both apoptosis and DNA release. By gel electrophoresis, extracellular DNA from apoptotic cells showed laddering with low molecular weight fragments. These studies suggest that extracellular release of DNA is a consequence of apoptosis and may account for some of the DNA in the blood.

Authors
Choi, J-J; Reich, CF; Pisetsky, DS
MLA Citation
Choi, J-J, Reich, CF, and Pisetsky, DS. "Release of DNA from dead and dying lymphocyte and monocyte cell lines in vitro." Scand J Immunol 60.1-2 (July 2004): 159-166.
PMID
15238085
Source
pubmed
Published In
Scandinavian Journal of Immunology
Volume
60
Issue
1-2
Publish Date
2004
Start Page
159
End Page
166
DOI
10.1111/j.0300-9475.2004.01470.x

Systemic lupus erythematosus and related diseases.

Authors
Tran, TT; Pisetsky, DS
MLA Citation
Tran, TT, and Pisetsky, DS. "Systemic lupus erythematosus and related diseases." Autoimmunity 37.4 (June 2004): 301-304. (Review)
PMID
15518046
Source
pubmed
Published In
Autoimmunity (Informa)
Volume
37
Issue
4
Publish Date
2004
Start Page
301
End Page
304

The role of biomechanics and inflammation in cartilage injury and repair.

Osteoarthritis is a painful and debilitating disease characterized by progressive degenerative changes in the articular cartilage and other joint tissues. Biomechanical factors play a critical role in the initiation and progression of this disease, as evidenced by clinical and animal studies of alterations in the mechanical environment of the joint caused by trauma, joint instability, disuse, or obesity. The onset of these changes after joint injury generally has been termed posttraumatic arthritis and can be accelerated by factors such as a displaced articular fracture. Within this context, there is considerable evidence that interactions between biomechanical factors and proinflammatory mediators are involved in the progression of cartilage degeneration in posttraumatic arthritis. In vivo studies have shown increased concentrations of inflammatory cytokines and mediators in the joint in mechanically induced models of osteoarthritis. In vitro explant studies confirm that mechanical load is a potent regulator of matrix metabolism, cell viability, and the production of proinflammatory mediators such as nitric oxide and prostaglandin E2. Knowledge of the interaction of inflammatory and biomechanical factors in regulating cartilage metabolism would be beneficial to an understanding of the etiopathogenesis of posttraumatic osteoarthritis and in the improvement of therapies for joint injury.

Authors
Guilak, F; Fermor, B; Keefe, FJ; Kraus, VB; Olson, SA; Pisetsky, DS; Setton, LA; Weinberg, JB
MLA Citation
Guilak, F, Fermor, B, Keefe, FJ, Kraus, VB, Olson, SA, Pisetsky, DS, Setton, LA, and Weinberg, JB. "The role of biomechanics and inflammation in cartilage injury and repair." Clin Orthop Relat Res 423 (June 2004): 17-26. (Review)
PMID
15232421
Source
pubmed
Published In
Clinical Orthopaedics and Related Research ®
Issue
423
Publish Date
2004
Start Page
17
End Page
26

Induction of immune activation by a novel immunomodulatory oligonucleotide without thymocyte apoptosis.

Bacterial DNA and synthetic oligodeoxynucleotides (ODN) containing unmethylated CpG motifs (CpG DNA) can potently stimulate innate immunity. While the actions of CpG DNA resemble those of LPS, these molecules stimulate distinct Toll-like receptors as well as cell types. In a previous study, we showed that a CpG ODN could induce cytokine production but, unlike LPS, did not induce thymocyte apoptosis. In this study, we have further investigated these differences using as a model a second-generation immunostimulatory oligonucleotide called HYB2048. Following administration to normal BALB/c mice, HYB2048-induced IL-12 but not IL-6 production. Under conditions in which LPS induced thymocyte apoptosis, HYB2048 did not cause significant cell death and, furthermore, did not block apoptosis induced by LPS. The levels of corticosterone induced by HYB2048 were also significantly lower than those induced by LPS. This pattern of activation could distinguish CpG DNA from LPS in its effects on the immune system.

Authors
Jiang, W; Reich, CF; You, D; Kandimalla, E; Agrawal, S; Pisetsky, DS
MLA Citation
Jiang, W, Reich, CF, You, D, Kandimalla, E, Agrawal, S, and Pisetsky, DS. "Induction of immune activation by a novel immunomodulatory oligonucleotide without thymocyte apoptosis." Biochem Biophys Res Commun 318.1 (May 21, 2004): 60-66.
PMID
15110753
Source
pubmed
Published In
Biochemical and Biophysical Research Communications
Volume
318
Issue
1
Publish Date
2004
Start Page
60
End Page
66
DOI
10.1016/j.bbrc.2004.04.001

The effect of dexamethasone on the generation of plasma DNA from dead and dying cells.

To determine the effects of glucocorticoids on the clearance of apoptotic and necrotic cells, the influence of dexamethasone on plasma levels of DNA was assessed in BALB/c mice receiving Jurkat cells treated with etoposide or ethanol. In untreated mice, administration of 10(8) apoptotic or necrotic Jurkat cells led to the appearance of DNA in the plasma. In mice treated 24 hours previously with dexamethasone, levels of DNA were reduced in a dose-dependent manner, with mice receiving 1 and 2.5 mg showing no appreciable plasma DNA levels. Similar results were obtained with assay of lactate dehydrogenase in mice receiving apoptotic cells. The effects of dexamethasone on anti-Fas treatment were also characterized. While treatment with a monoclonal anti-Fas reagent caused a significant plasma DNA response in untreated mice, mice pretreated with dexamethasone showed much lower levels. Blood levels of caspase 3 and TUNEL staining of liver were also reduced in dexamethasone-treated mice compared to controls receiving anti-Fas antibody. These results indicate that glucocorticoids can affect the clearance of apoptotic and necrotic cells as well as the induction of apoptosis in at least some tissues. These activities may be relevant to the efficacy of glucocorticoids in the treatment of inflammatory disease.

Authors
Jiang, N; Pisetsky, DS
MLA Citation
Jiang, N, and Pisetsky, DS. "The effect of dexamethasone on the generation of plasma DNA from dead and dying cells." Am J Pathol 164.5 (May 2004): 1751-1759.
PMID
15111321
Source
pubmed
Published In
The American journal of pathology
Volume
164
Issue
5
Publish Date
2004
Start Page
1751
End Page
1759
DOI
10.1016/S0002-9440(10)63733-9

Mechanisms of activation of the RAW264.7 macrophage cell line by transfected mammalian DNA.

Bacterial DNA can stimulate the production of cytokines and nitric oxide (NO), while mammalian DNA can block these responses. If mammalian DNA is transfected into macrophages, however, it can stimulate NO production, without inducing IL-12. To define further this activity, signaling pathways induced by transfected calf thymus (CT) DNA were studied. Using RAW264.7 cells as a model, CT DNA in the transfection agent FuGENE 6 activated cells through the NF-kappaB and MAPKs pathways, similar to bacterial DNA and LPS. The role of these pathways was further investigated using specific inhibitors, with studies indicating that NO production is blocked by inhibitors of NF-kappaB and p38 but not other MAPKs. These data indicate that the immune activity of DNA is influenced by context or intracellular location and that, when transfected into cells, mammalian DNA can activate cells through signaling pathways similar to those of bacterial DNA.

Authors
Jiang, W; Reich III, CF; Pisetsky, DS
MLA Citation
Jiang, W, Reich III, CF, and Pisetsky, DS. "Mechanisms of activation of the RAW264.7 macrophage cell line by transfected mammalian DNA." Cell Immunol 229.1 (May 2004): 31-40.
PMID
15331326
Source
pubmed
Published In
Cellular Immunology
Volume
229
Issue
1
Publish Date
2004
Start Page
31
End Page
40
DOI
10.1016/j.cellimm.2004.06.003

OutOfDate.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "OutOfDate." Pharos Alpha Omega Alpha Honor Med Soc 67.3 (2004): 17-18.
PMID
15449885
Source
pubmed
Published In
The Pharos of Alpha Omega Alpha-Honor Medical Society. Alpha Omega Alpha
Volume
67
Issue
3
Publish Date
2004
Start Page
17
End Page
18

Apoptosis in the rheumatic diseases

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Apoptosis in the rheumatic diseases." Rheumatic Disease Clinics of North America 30.3 (2004): xiii-xiv.
Source
scival
Published In
Rheumatic Disease Clinics of North America
Volume
30
Issue
3
Publish Date
2004
Start Page
xiii
End Page
xiv
DOI
10.1016/j.rdc.2004.05.001

Specificity and immunochemical properties of anti-DNA antibodies induced in normal mice by immunization with mammalian DNA with a CpG oligonucleotide as adjuvant.

To elucidate the role of DNA antigen drive in the anti-DNA response, the specificity and immunochemical properties of anti-DNA antibodies induced in normal mice by immunization with double stranded (ds) mammalian DNA with a CpG oligonucleotide (ODN) adjuvant were characterized. Like spontaneous anti-DNA from MRL/lpr mice, the induced anti-DNA bound cross-reactively to DNA from five different species by ELISA. The induced antibodies displayed a predominance of IgG2a and had much lower amount of IgG3 than spontaneous antibodies. Surface plasmon resonance indicated that the induced and spontaneous anti-DNA antibodies have a similar range of avidity and binding kinetics. While sera from the MRL/lpr mice had substantial binding to histones and nucleosomes, the immunized mice had antibody levels to these antigens similar to those of mice treated only with incomplete Freund's adjuvant. Together, these results indicate that normal mice can produce autoantibodies to dsDNA, with a CpG ODN allowing the generation of antibodies resembling those in spontaneous autoimmunity.

Authors
Tran, TT; Reich, CF; Alam, M; Pisetsky, DS
MLA Citation
Tran, TT, Reich, CF, Alam, M, and Pisetsky, DS. "Specificity and immunochemical properties of anti-DNA antibodies induced in normal mice by immunization with mammalian DNA with a CpG oligonucleotide as adjuvant." Clin Immunol 109.3 (December 2003): 278-287.
PMID
14697742
Source
pubmed
Published In
Clinical Immunology
Volume
109
Issue
3
Publish Date
2003
Start Page
278
End Page
287

Role of macrophages in the generation of circulating blood nucleosomes from dead and dying cells.

After apoptosis or necrosis, macrophages clear dead cells by phagocytosis. Although this process is efficient, circulating nucleosomes can occur in certain diseases, presumably reflecting either increased production or impaired clearance. To investigate the generation of blood nucleosomes, graded numbers of apoptotic and necrotic cells were administered to healthy mice, and levels of blood nucleosomes and DNA were determined. Using Jurkat cells as a model, nucleosomes and DNA were detected in the blood after the administration of 108 apoptotic or necrotic cells per mouse by the intraperitoneal route. The kinetics of the response were similar for both types of cells. The role of macrophages was assessed by eliminating these cells with clodronate liposomes or silica. Although clodronate treatment alone produced a peak level of blood DNA, the subsequent administration of dead cells caused no change in DNA levels. In contrast, silica treatment alone did not elicit a blood DNA response, though this treatment limited the rise in DNA from administered cells. Molecular studies showed that the blood DNA following the administration of apoptotic or necrotic cells arose from the mouse and the Jurkat cells, and its size distribution was consistent with apoptosis. Together, these findings suggest that the generation of blood nucleosomes depends on macrophages, with apoptosis a concomitant of a high burden of dead and dying cells.

Authors
Jiang, N; Reich, CF; Pisetsky, DS
MLA Citation
Jiang, N, Reich, CF, and Pisetsky, DS. "Role of macrophages in the generation of circulating blood nucleosomes from dead and dying cells." Blood 102.6 (September 15, 2003): 2243-2250.
PMID
12775567
Source
pubmed
Published In
Blood
Volume
102
Issue
6
Publish Date
2003
Start Page
2243
End Page
2250
DOI
10.1182/blood-2002-10-3312

B lymphocytes and systemic lupus erythematosus.

Systemic lupus erythematosus (SLE) is a prototypic autoimmune disease characterized by B cell hyperactivity in association with autoantibodies, most prominently those directed to components of the cell nucleus. The source of the antigens that drive B cell responses in SLE is unknown, although recent studies suggest mechanisms by which the self-antigens become immunogenic and stimulate responses. Among these mechanisms, abnormalities in the generation of apoptotic cells or their clearance may increase the availability of nuclear antigens to drive responses. In addition, autoantibody crossreactivity may promote induction of responses to disparate antigens, foreign and self, and enable a single autoantibody to cause disease by crossreactive binding. In addition to reflecting increased exposure to self-antigen, autoantibody responses in SLE may result from abnormalities in B cell signaling and regulation by cytokines. New approaches to therapy aim to abrogate autoantibody production by targeting specific steps in B cell activation, including blockade of T cell costimulation.

Authors
Criscione, LG; Pisetsky, DS
MLA Citation
Criscione, LG, and Pisetsky, DS. "B lymphocytes and systemic lupus erythematosus." Curr Rheumatol Rep 5.4 (August 2003): 264-269. (Review)
PMID
14531953
Source
pubmed
Published In
Current Rheumatology Reports
Volume
5
Issue
4
Publish Date
2003
Start Page
264
End Page
269

Regulation of matrix turnover in meniscal explants: role of mechanical stress, interleukin-1, and nitric oxide.

The meniscus is an intra-articular fibrocartilaginous structure that serves essential biomechanical roles in the knee. With injury or arthritis, the meniscus may be exposed to significant changes in its biochemical and biomechanical environments that likely contribute to the progression of joint disease. The goal of this study was to examine the influence of mechanical stress on matrix turnover in the meniscus in the presence of interleukin-1 (IL-1) and to determine the role of nitric oxide (NO) in these processes. Explants of porcine menisci were subjected to dynamic compressive stresses at 0.1 MPa for 24 h at 0.5 Hz with 1 ng/ml IL-1, and the synthesis of total protein, proteoglycan, and NO was measured. The effects of a nitric oxide synthase 2 (NOS2) inhibitor were determined. Dynamic compression significantly increased protein and proteoglycan synthesis by 68 and 58%, respectively, compared with uncompressed explants. This stimulatory effect of mechanical stress was prevented by the presence of IL-1 but was restored by specifically inhibiting NOS2. Release of proteoglycans into the medium was increased by IL-1 or mechanical compression and further enhanced by IL-1 and compression together. Stimulation of proteoglycan release in response to compression was dependent on NOS2 regardless of the presence of IL-1. These finding suggest that IL-1 may modulate the effects of mechanical stress on extracellular matrix turnover through a pathway that is dependent on NO.

Authors
Shin, S-J; Fermor, B; Weinberg, JB; Pisetsky, DS; Guilak, F
MLA Citation
Shin, S-J, Fermor, B, Weinberg, JB, Pisetsky, DS, and Guilak, F. "Regulation of matrix turnover in meniscal explants: role of mechanical stress, interleukin-1, and nitric oxide." J Appl Physiol (1985) 95.1 (July 2003): 308-313.
PMID
12665533
Source
pubmed
Published In
Journal of applied physiology (Bethesda, Md. : 1985)
Volume
95
Issue
1
Publish Date
2003
Start Page
308
End Page
313
DOI
10.1152/japplphysiol.00131.2003

Informed consent in a clinical trial of a novel treatment for rheumatoid arthritis.

OBJECTIVE: To evaluate the informed consent process for a clinical trial of intravenous doxycycline for rheumatoid arthritis. METHODS: Participants completed a self-administered questionnaire about the consent process at baseline and 16 weeks following enrollment in a clinical trial. RESULTS: Respondents (n = 30) affirmed voluntary participation in the parent trial. Participants acknowledged hope and altruism as reasons for entering the trial more than expectation of personal benefit or outside influences. Many respondents did not understand randomization (14/30), placebos (15/30), or risks of study medications; 11/30 respondents believed that the study drug was completely safe. CONCLUSION: Respondents generally understood the experimental nature of the trial and confirmed their participation was voluntary. However, gaps existed in participants understanding of trial design, raising the question of whether they were adequately informed about the research study prior to enrollment. Further education of potential participants in clinical trials may be required to achieve valid informed consent.

Authors
Criscione, LG; Sugarman, J; Sanders, L; Pisetsky, DS; St Clair, EW
MLA Citation
Criscione, LG, Sugarman, J, Sanders, L, Pisetsky, DS, and St Clair, EW. "Informed consent in a clinical trial of a novel treatment for rheumatoid arthritis." Arthritis Rheum 49.3 (June 15, 2003): 361-367.
PMID
12794792
Source
pubmed
Published In
Arthritis and Rheumatism
Volume
49
Issue
3
Publish Date
2003
Start Page
361
End Page
367
DOI
10.1002/art.11057

Effect of cytofectins on the immune response of murine macrophages to mammalian DNA.

DNA, depending on base sequence, can induce a wide range of immune responses. While bacterial DNA is stimulatory, mammalian DNA is inactive alone and can, moreover, inhibit the response to bacterial DNA. To determine whether the mode of cell entry affects the immune properties of mammalian DNA, we have investigated the effects of the cytofectin agents Fugene 6 (Roche Diagnostics Corp., Indianapolis, IN), Lipofectin and Lipofectamine (Life Technologies, Grand Island, NY) on the responses of murine macrophages to DNA from calf thymus and human placenta. Whereas calf thymus and human placenta DNA alone failed to stimulate J774 or RAW264.7 cell lines or bone marrow-derived macrophages, these DNAs in complexes with cytofectin agents stimulated macrophages to produce nitric oxide but not interleukin 12. Both single-stranded and double-stranded DNAs were active in the presence of cytofectins. Macrophage activation by the DNA-cytofectin complexes was reduced by chloroquine, suggesting a role of endosomal acidification in activation. As shown by flow cytometry and confocal microscopy, the cytofectins caused an increase in the uptake of DNA into cells. Our findings indicate that macrophages vary in their response to DNA depending on uptake pathway, suggesting that activation by DNA reflects not only sequence but also context or intracellular location.

Authors
Zhu, F-G; Reich, CF; Pisetsky, DS
MLA Citation
Zhu, F-G, Reich, CF, and Pisetsky, DS. "Effect of cytofectins on the immune response of murine macrophages to mammalian DNA." Immunology 109.2 (June 2003): 255-262.
PMID
12757621
Source
pubmed
Published In
Immunology
Volume
109
Issue
2
Publish Date
2003
Start Page
255
End Page
262

The use of fluorometric assays to assess the immune response to DNA in murine systemic lupus erythematosus.

Antibodies to DNA (anti-DNA) play an important role in the pathogenesis of systemic lupus erythematosus (SLE). In blood, these antibodies may exist in a free, unbound state or as part of complexes with DNA. Furthermore, circulating DNA may be either complexed or free. Because of the central role of these immunoreactants (anti-DNA and DNA) in the disease, monitoring of their levels could provide valuable information for both clinical and investigative purposes. In these studies, we have explored the use of a DNA-binding dye, PicoGreen, for the detection of circulating DNA, either total or immune complex bound. In addition, we have used this dye for Farr-type antibody assays. Using autoimmune MRL/lpr mice as a model, we have shown that, while the levels of free DNA in the plasma of these mice were comparable with those of normal BALB/c mice, the amounts in complexes precipitable by ammonium sulfate were significantly greater. Furthermore, we showed that Farr assays using PicoGreen reliably detect levels of free anti-DNA, with values correlated with anti-DNA levels by an enzyme-linked immunosorbent assay. Together, our results suggest that a fluorometric dye can accurately monitor DNA and anti-DNA antibody levels in SLE and may provide important information on immunopathogenesis.

Authors
Björkman, L; Reich, CF; Pisetsky, DS
MLA Citation
Björkman, L, Reich, CF, and Pisetsky, DS. "The use of fluorometric assays to assess the immune response to DNA in murine systemic lupus erythematosus." Scand J Immunol 57.6 (June 2003): 525-533.
PMID
12791090
Source
pubmed
Published In
Scandinavian Journal of Immunology
Volume
57
Issue
6
Publish Date
2003
Start Page
525
End Page
533

New policy on disclosure of interest for American College of Rheumatology Journals.

Authors
Pisetsky, DS; Hunder, GG; Gravallese, EM
MLA Citation
Pisetsky, DS, Hunder, GG, and Gravallese, EM. "New policy on disclosure of interest for American College of Rheumatology Journals." Arthritis Rheum 49.2 (April 15, 2003): 149-150.
PMID
12687503
Source
pubmed
Published In
Arthritis and Rheumatism
Volume
49
Issue
2
Publish Date
2003
Start Page
149
End Page
150
DOI
10.1002/art.10992

Enhancing immunogenicity by CpG DNA.

Bacterial DNA and oligonucleotides containing unmethylated CpG dinucleotides (CpG DNA) can stimulate immune responses and have potential for use as novel agents to enhance immunogenicity. CpG DNA can interact with toll-like receptor 9 and cause activation through a myeloid differentiation primary response gene (MyD88)-dependent signaling pathway. Due to its pattern of immune cell activation, CpG DNA can induce a cytokine milieu to promote T-helper cell responses and serve as an adjuvant. Furthermore, CpG DNA can provide protection against pathogens in animal models and has therapeutic applications in clinical settings such as in cancer and allergy.

Authors
Jiang, W; Pisetsky, DS
MLA Citation
Jiang, W, and Pisetsky, DS. "Enhancing immunogenicity by CpG DNA." Curr Opin Mol Ther 5.2 (April 2003): 180-185. (Review)
PMID
12772509
Source
pubmed
Published In
Current Opinion in Molecular Therapeutics
Volume
5
Issue
2
Publish Date
2003
Start Page
180
End Page
185

New policy on disclosure of interest for American College of Rheumatology journals.

Authors
Pisetsky, DS; Hunder, GG; Gravallese, EM
MLA Citation
Pisetsky, DS, Hunder, GG, and Gravallese, EM. "New policy on disclosure of interest for American College of Rheumatology journals." Arthritis Rheum 48.4 (April 2003): 873-875.
PMID
12687527
Source
pubmed
Published In
Arthritis and Rheumatism
Volume
48
Issue
4
Publish Date
2003
Start Page
873
End Page
875
DOI
10.1002/art.10946

Screening the genome for rheumatoid arthritis susceptibility genes: a replication study and combined analysis of 512 multicase families.

OBJECTIVE: A number of non-HLA loci that have shown evidence (P < 0.05) for linkage with rheumatoid arthritis (RA) have been previously identified. The present study attempts to confirm these findings. METHODS: We performed a second genome-wide screen of 256 new multicase RA families recruited from across the United States by the North American Rheumatoid Arthritis Consortium. Affected sibling pair analysis on the new data set was performed using SIBPAL. We subsequently combined our first and second data sets in an attempt to enhance the evidence for linkages in a larger sample size. We also evaluated the impact of covariates on the support for linkage, using LODPAL. RESULTS: Evidence of linkage at 1p13 (D1S1631), 6p21.3 (the HLA complex), and 18q21 (D18S858) (P < 0.05) was replicated in this independent data set. In addition, there was new evidence for linkage at 9p22 (D9S1121 [P = 0.001]) and 10q21 (D10S1221 [P = 0.0002] and D10S1225 [P = 0.0038]) in the current data set. The combined analysis of both data sets (512 families) showed evidence for linkage at the level of P < 0.005 at 1p13 (D1S1631), 1q43 (D1S235), 6q21 (D6S2410), 10q21 (D10S1221), 12q12 (D12S398), 17p13 (D17S1298), and 18q21 (D18S858). Linkage at HLA was also confirmed (P < 5 x 10(-12)). Inclusion of DRB1*04 as a covariate significantly increased the probability of linkage on chromosome 6. In addition, some linkages on chromosome 1 showed improved significance when modeling DRB1*04 or rheumatoid factor positivity as covariates. CONCLUSION: These results provide a rational basis for pursuing high-density linkage and association studies of RA in several regions outside of the HLA region, particularly on chromosomes 1p, 1q, and 18q.

Authors
Jawaheer, D; Seldin, MF; Amos, CI; Chen, WV; Shigeta, R; Etzel, C; Damle, A; Xiao, X; Chen, D; Lum, RF; Monteiro, J; Kern, M; Criswell, LA; Albani, S; Nelson, JL; Clegg, DO; Pope, R; Schroeder, HW; Bridges, SL; Pisetsky, DS; Ward, R; Kastner, DL; Wilder, RL; Pincus, T; Callahan, LF; Flemming, D; Wener, MH; Gregersen, PK; North American Rheumatoid Arthritis Consortium,
MLA Citation
Jawaheer, D, Seldin, MF, Amos, CI, Chen, WV, Shigeta, R, Etzel, C, Damle, A, Xiao, X, Chen, D, Lum, RF, Monteiro, J, Kern, M, Criswell, LA, Albani, S, Nelson, JL, Clegg, DO, Pope, R, Schroeder, HW, Bridges, SL, Pisetsky, DS, Ward, R, Kastner, DL, Wilder, RL, Pincus, T, Callahan, LF, Flemming, D, Wener, MH, Gregersen, PK, and North American Rheumatoid Arthritis Consortium, . "Screening the genome for rheumatoid arthritis susceptibility genes: a replication study and combined analysis of 512 multicase families." Arthritis Rheum 48.4 (April 2003): 906-916.
PMID
12687532
Source
pubmed
Published In
Arthritis and Rheumatism
Volume
48
Issue
4
Publish Date
2003
Start Page
906
End Page
916
DOI
10.1002/art.10989

The expression of plasma nucleosomes in mice undergoing in vivo apoptosis.

Nucleosomes occur in the blood of patients with systemic lupus erythematosus and are thought to result from in vivo cell death. To determine the conditions for the release of nucleosomes into the blood, normal mice were treated with four agents that have the potential to induce apoptosis or immune cell activation in vivo: LPS, CpG DNA, anti-Fas antibody, and dexamethasone. Blood nucleosomes were measured by a capture ELISA immunoassay, with the DNA component assessed by fluorimetry with the dye PicoGreen. Following treatment with LPS and a monoclonal anti-Fas antibody, nucleosomes and DNA appeared in the plasma in a dose-dependent fashion. In contrast, dexamethasone treatment, despite causing significant thymocyte loss, did not elicit plasma nucleosomes. Similarly, CpG DNA, while inducing an IL-12 response comparable to that of LPS, also did not elicit plasma nucleosomes. These results suggest that plasma nucleosome levels reflect specific patterns of cell death and are not an invariable consequence of in vivo apoptosis or immune cell activation.

Authors
Jiang, N; Reich, CF; Monestier, M; Pisetsky, DS
MLA Citation
Jiang, N, Reich, CF, Monestier, M, and Pisetsky, DS. "The expression of plasma nucleosomes in mice undergoing in vivo apoptosis." Clin Immunol 106.2 (February 2003): 139-147.
PMID
12672404
Source
pubmed
Published In
Clinical Immunology
Volume
106
Issue
2
Publish Date
2003
Start Page
139
End Page
147

Etanercept-induced lupus-like syndrome in a patient with rheumatoid arthritis [6] (multiple letters)

Authors
Carlson, E; Rothfield, N; Pisetsky, DS
MLA Citation
Carlson, E, Rothfield, N, and Pisetsky, DS. "Etanercept-induced lupus-like syndrome in a patient with rheumatoid arthritis [6] (multiple letters)." Arthritis and Rheumatism 48.4 (2003): 1165-1166.
PMID
12687569
Source
scival
Published In
Arthritis and Rheumatism
Volume
48
Issue
4
Publish Date
2003
Start Page
1165
End Page
1166
DOI
10.1002/art.11033

Minimum Information About a Microarray Experiment: Comment on the editorial by Firestein and Pisetsky [5]

Authors
Distler, O; Gay, S; Neumann, E; Müller-Ladner, U; Firestein, GS; Pisetsky, DS
MLA Citation
Distler, O, Gay, S, Neumann, E, Müller-Ladner, U, Firestein, GS, and Pisetsky, DS. "Minimum Information About a Microarray Experiment: Comment on the editorial by Firestein and Pisetsky [5]." Arthritis and Rheumatism 48.3 (2003): 861-862.
PMID
12632450
Source
scival
Published In
Arthritis and Rheumatism
Volume
48
Issue
3
Publish Date
2003
Start Page
861
End Page
862
DOI
10.1002/art.10756

The pathogenesis of systemic lupus erythematosus

Authors
Criscione, LG; Pisetsky, DS
MLA Citation
Criscione, LG, and Pisetsky, DS. "The pathogenesis of systemic lupus erythematosus." Bulletin on the Rheumatic Diseases 52.6 (2003).
Source
scival
Published In
Bulletin on the rheumatic diseases
Volume
52
Issue
6
Publish Date
2003

Inhibition of murine dendritic cell activation by synthetic phosphorothioate oligodeoxynucleotides.

Depending on sequence and backbone structure, DNA can inhibit as well as stimulate immune responses. As previously shown, single-base phosphorothioate (Ps) oligodeoxynucleotides (ODN) can inhibit murine macrophage activation. To determine whether these compounds can also affect dendritic cells (DC), the effects of 30-mer Ps ODN (SdA, SdT, SdG, and SdC) on DC activation were assessed in an in vitro system. With DC preparations obtained from murine bone marrow cultured in granulocyte macrophage-colony stimulating factor, the Ps ODN blocked the production of interleukin-12 and nitric oxide induced by bacterial DNA, an immunostimulatory cytosine phosphate guanosine dinucleotide (CpG) ODN and lipopolysaccharide (LPS). Furthermore, these compounds inhibited up-regulation of costimulatory molecules CD40 and CD86 as well as major histocompatibility complex-II molecules, indicating an effect on DC maturation. Although the Ps ODN limited uptake of CpG ODN as assessed by flow cytometry, the Ps ODN did not affect LPS uptake, suggesting that these compounds inhibit DC responses by effects on downstream signaling pathways. Together, these observations extend the range of action of inhibitory ODN to DC and suggest a role of these compounds as immunomodulatory agents.

Authors
Zhu, F-G; Reich, CF; Pisetsky, DS
MLA Citation
Zhu, F-G, Reich, CF, and Pisetsky, DS. "Inhibition of murine dendritic cell activation by synthetic phosphorothioate oligodeoxynucleotides." J Leukoc Biol 72.6 (December 2002): 1154-1163.
PMID
12488497
Source
pubmed
Published In
Journal of leukocyte biology
Volume
72
Issue
6
Publish Date
2002
Start Page
1154
End Page
1163

Induction of cyclooxygenase-2 by mechanical stress through a nitric oxide-regulated pathway.

OBJECTIVE: Biomechanical signals play important roles in regulating the homeostasis of articular cartilage, but under abnormal conditions may be a critical factor in the onset and progression of arthritis. Prostaglandin E(2) (PGE(2)) and nitric oxide (NO), derived from the enzymes cyclo-oxygenase 2 (COX2) and NO synthase 2 (NOS2), are inflammatory mediators that modulate numerous physiological and pathophysiological processes and are potentially important pharmacological targets in osteoarthritis. The goal of this study was to determine the effect of mechanical compression on PGE(2) production in the presence of selective NOS2 and COX2 inhibitors. METHODS: Articular cartilage explants harvested from 2-3-year-old pigs were subjected to intermittent compression at 0.5Hz over a range of stress magnitudes. PGE(2) and NO production into the media were determined in the presence and absence of the NOS2 inhibitor 1400W or the COX2 inhibitor NS398. COX2 protein levels were determined by immunoblot analysis. RESULTS: Mechanical compression significantly increased NO and PGE(2) synthesis in a manner that was dependent on the magnitude of stress. The selective COX2 inhibitor blocked compression-induced NO and PGE(2) production. Compression in the presence of 1400W further increased COX2 expression resulting in a 10-fold increase in PGE(2) production compared to uncompressed explants with 1400W and a 40-fold increase in PGE(2) compared to uncompressed explants without 1400W. CONCLUSION: Mechanical compression of articular cartilage increased COX2 and PGE(2) production through a NO-dependent pathway, and therefore pharmacological agents that target the NOS2 pathway in cartilage may have a significant influence on prostanoid production in the joint.

Authors
Fermor, B; Weinberg, JB; Pisetsky, DS; Misukonis, MA; Fink, C; Guilak, F
MLA Citation
Fermor, B, Weinberg, JB, Pisetsky, DS, Misukonis, MA, Fink, C, and Guilak, F. "Induction of cyclooxygenase-2 by mechanical stress through a nitric oxide-regulated pathway." Osteoarthritis Cartilage 10.10 (October 2002): 792-798.
PMID
12359165
Source
pubmed
Published In
Osteoarthritis and Cartilage
Volume
10
Issue
10
Publish Date
2002
Start Page
792
End Page
798

A walk on the beach.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "A walk on the beach." Ann Intern Med 137.5 Part 1 (September 3, 2002): 366-367.
PMID
12204025
Source
pubmed
Published In
Annals of internal medicine
Volume
137
Issue
5 Part 1
Publish Date
2002
Start Page
366
End Page
367

DNA microarrays: boundless technology or bound by technology? Guidelines for studies using microarray technology.

Authors
Firestein, GS; Pisetsky, DS
MLA Citation
Firestein, GS, and Pisetsky, DS. "DNA microarrays: boundless technology or bound by technology? Guidelines for studies using microarray technology." Arthritis Rheum 46.4 (April 2002): 859-861. (Review)
PMID
11953960
Source
pubmed
Published In
Arthritis and Rheumatism
Volume
46
Issue
4
Publish Date
2002
Start Page
859
End Page
861

Inhibition of murine macrophage nitric oxide production by synthetic oligonucleotides.

Synthetic 30-mer phosphorothioate (Ps) oligonucleotides (ODN) comprised of single bases (SdA30, SdC30, SdG30, and SdT30) were assessed for their effects on nitric oxide (NO) production by murine bone marrow macrophages (BMMC) and macrophage cell lines J774 and RAW264.7. Pretreatment of these cells with any of the four Ps ODN inhibited NO production induced by CpG ODN, E. coli DNA (EC DNA), or LPS. This inhibition was time- and dose-dependent and was observed even if the Ps ODN were added as long as 12 h after stimulation. As in the case of stimulatory ODN, inhibition was dependent on backbone structure and length. Thus, all four 30-mer, single-base Ps ODN were inhibitory, and only dG30 among phosphodiester ODN was inhibitory. Together, these observations indicate that Ps ODN can inhibit macrophage production of inflammatory mediators, suggesting a role of these compounds as immunomodulatory agents.

Authors
Zhu, F-G; Reich, CF; Pisetsky, DS
MLA Citation
Zhu, F-G, Reich, CF, and Pisetsky, DS. "Inhibition of murine macrophage nitric oxide production by synthetic oligonucleotides." J Leukoc Biol 71.4 (April 2002): 686-694.
PMID
11927656
Source
pubmed
Published In
Journal of leukocyte biology
Volume
71
Issue
4
Publish Date
2002
Start Page
686
End Page
694

Influence of hypoxia and reoxygenation on cytokine-induced production of proinflammatory mediators in articular cartilage.

OBJECTIVE: Articular cartilage is an avascular tissue that functions at a lower oxygen tension than do most tissues. With mobilization, arthritic joints may undergo cycles of hypoxia and reoxygenation. The goal of this study was to determine the effects of hypoxia and reoxygenation on cytokine-induced nitric oxide (NO) and prostaglandin E(2) (PGE(2)) production in articular cartilage. METHODS: Porcine cartilage explants were incubated at 37 degrees C for 72 hours in either 1% O(2) (hypoxia) or 20% O(2) (normoxia) in media supplemented with interleukin-1alpha (IL-1alpha) or tumor necrosis factor alpha (TNFalpha), with or without the NO synthase 2 (NOS2) selective inhibitor 1400W. Culture media were then removed and replaced with freshly prepared media and incubated for a further 24 hours in normoxia. RESULTS: NO levels were significantly higher in explants supplemented with IL-1alpha and TNFalpha compared with controls, in both hypoxia and normoxia. Compared with normoxia, hypoxia decreased IL-1alpha- and TNFalpha-induced NO production significantly. Reoxygenation of hypoxic explants resulted in sustained significant NO production in response to either cytokine. However, comparably high levels of NO production were not sustained in explants cultured continuously in normoxia. Although IL-1alpha alone did not significantly increase PGE(2) production, significant PGE(2) superinduction occurred in cartilage stimulated with IL-1alpha and the NOS2 inhibitor 1400W compared with stimulation with IL-1alpha alone in hypoxia, but not in normoxia. CONCLUSION: Oxygen tension significantly affects cytokine-induced proinflammatory mediator production in articular cartilage. Furthermore, hypoxia alters NO mediation of PGE(2) production. Hypoxia and reoxygenation can affect cytokine-induced proinflammatory mediator production, suggesting that oxygen tension may influence inflammation associated with cartilage injury and disease.

Authors
Cernanec, J; Guilak, F; Weinberg, JB; Pisetsky, DS; Fermor, B
MLA Citation
Cernanec, J, Guilak, F, Weinberg, JB, Pisetsky, DS, and Fermor, B. "Influence of hypoxia and reoxygenation on cytokine-induced production of proinflammatory mediators in articular cartilage." Arthritis Rheum 46.4 (April 2002): 968-975.
PMID
11953974
Source
pubmed
Published In
Arthritis and Rheumatism
Volume
46
Issue
4
Publish Date
2002
Start Page
968
End Page
975

A medical education.

Authors
Pisetsky, D
MLA Citation
Pisetsky, D. "A medical education." Lancet 359.9306 (2002): 636--.
PMID
11867161
Source
scival
Published In
Lancet
Volume
359
Issue
9306
Publish Date
2002
Start Page
636-

Progress in the treatment of rheumatoid arthritis.

Authors
Pisetsky, DS; St Clair, EW
MLA Citation
Pisetsky, DS, and St Clair, EW. "Progress in the treatment of rheumatoid arthritis." JAMA 286.22 (December 12, 2001): 2787-2790. (Review)
PMID
11735734
Source
pubmed
Published In
JAMA : the journal of the American Medical Association
Volume
286
Issue
22
Publish Date
2001
Start Page
2787
End Page
2790

The role of surface ig binding in the activation of human B cells by phosphorothioate oligodeoxynucleotides.

Phosphorothioate oligodeoxynucleotides (sODNs) can induce T-cell-independent polyclonal activation of human B cells by a mechanism that depends on both sequence and back-bone structure. Because matrix-bound as well as soluble sODNs are mitogenic, this stimulation may result from the engagement of surface receptor(s). In order to investigate whether surface immunoglobin (Ig) could be a receptor for sODNs, the interaction of sODNs-fluorescein isothiocyanate (FITC) with Ig-coated beads was examined. sODNs specifically bound to human IgM and IgG. Moreover, binding of sODN to human B cells induced temperature-dependent capping of bound receptors and colocalization of FITC-sODN and IgM into aggregated caps on the surface of human B cells. A role of surface Ig was furthermore shown by observations that antibody-mediated capping of B-cell surface IgM or IgD inhibited subsequent binding of sODNs and that the capacity of sODN to stimulate human B cells was blocked by excess IgM or IgG, by nonstimulatory antibodies to sIgM, as well as by a variety of negatively charged molecules. Together, these results indicate that sODNs engage surface Ig by charge-charge interactions that lead to activation of human B cells.

Authors
Liang, H; Reich, CF; Pisetsky, DS; Lipsky, PE
MLA Citation
Liang, H, Reich, CF, Pisetsky, DS, and Lipsky, PE. "The role of surface ig binding in the activation of human B cells by phosphorothioate oligodeoxynucleotides." Scand J Immunol 54.6 (December 2001): 551-563.
PMID
11902330
Source
pubmed
Published In
Scandinavian Journal of Immunology
Volume
54
Issue
6
Publish Date
2001
Start Page
551
End Page
563

Host response to infection: the role of CpG DNA in induction of cyclooxygenase 2 and nitric oxide synthase 2 in murine macrophages.

Depending on sequence, bacterial and synthetic DNAs can activate the host immune system and influence the host response to infection. The purpose of this study was to determine the abilities of various phosphorothioate oligonucleotides with cytosine-guanosine-containing motifs (CpG DNA) to activate macrophages to produce nitric oxide (NO) and prostaglandin E(2) (PGE(2)) and to induce expression of NO synthase 2 (NOS2) and cyclooxygenase 2 (COX2). As little as 0.3 microg of CpG DNA/ml increased NO and PGE(2) production in a dose- and time-dependent fashion in cells of the mouse macrophage cell line J774. NO and PGE(2) production was noted by 4 to 8 h after initiation of cultures with the CpG DNA, with the kinetics of NO production induced by CpG DNA being comparable to that induced by a combination of lipopolysaccharide and gamma interferon. CpG DNA-treated J774 cells showed enhanced expression of NOS2 and COX2 proteins as determined by immunoblotting, with the relative potencies of the CpG DNAs generally corresponding to those noted for the induction of NO and PGE(2) production as well as to those noted for the induction of interleukin-6 (IL-6), IL-12, and tumor necrosis factor. Extracts from CpG DNA-treated cells converted L-arginine to L-citrulline, but the NOS inhibitor N(G)-monomethyl-L-arginine (NMMA) inhibited this reaction. The COX2-specific inhibitor NS398 inhibited CpG DNA-induced PGE(2) production and inhibited NO production to various degrees. The NOS inhibitors NMMA, 1400W, and N-iminoethyl-L-lysine effectively blocked NO production and increased the production of PGE(2) in a dose-dependent fashion. Thus, analogues of microbial DNA (i.e., CpG DNA) activate mouse macrophage lineage cells for the expression of NOS2 and COX2, with the production of NO and that of PGE(2) occurring in an interdependent manner.

Authors
Ghosh, DK; Misukonis, MA; Reich, C; Pisetsky, DS; Weinberg, JB
MLA Citation
Ghosh, DK, Misukonis, MA, Reich, C, Pisetsky, DS, and Weinberg, JB. "Host response to infection: the role of CpG DNA in induction of cyclooxygenase 2 and nitric oxide synthase 2 in murine macrophages." Infect Immun 69.12 (December 2001): 7703-7710.
PMID
11705951
Source
pubmed
Published In
Infection and immunity
Volume
69
Issue
12
Publish Date
2001
Start Page
7703
End Page
7710
DOI
10.1128/IAI.69.12.7703-7710.2001

Immune response to DNA in systemic lupus erythematosus.

Antibodies to DNA occur prominently in systemic lupus erythematosus and have been extensively studied as probes for underlying immune disturbances. These antibodies have features of DNA antigen drive. While previous models for this response posited DNA as simple and inert, recent studies have indicated that DNA is immunologically diverse and, depending upon sequence and backbone structure, can stimulate or suppress immune responses. In particular, bacterial DNA is immunologically potent and can function as both an adjuvant and immunogen, eliciting in normal individuals antibodies to sites exclusive to bacterial DNA. In mice genetically predisposed to autoimmunity, however, bacterial DNA can elicit anti-DNA autoantibodies under conditions in which mammalian DNA is inactive. These findings suggest that foreign DNA can serve as a trigger for anti-DNA responses, with SLE reflecting a disturbance in antibody specificity and a shift from binding of sequential to backbone determinants. In contrast to bacterial DNA, mammalian DNA can suppress certain immune responses and prevent macrophage cytokine production. To the extent that self-DNA drives responses in SLE, anti-DNA production in this disease may reflect a failure of this suppression. The recognition of DNA's immune activities thus suggests novel possibilities for disease pathogenesis.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Immune response to DNA in systemic lupus erythematosus." Isr Med Assoc J 3.11 (November 2001): 850-853. (Review)
PMID
11729584
Source
pubmed
Published In
The Israel Medical Association journal : IMAJ
Volume
3
Issue
11
Publish Date
2001
Start Page
850
End Page
853

Role of the heat shock protein 90 in immune response stimulation by bacterial DNA and synthetic oligonucleotides.

To elucidate the mechanisms of immunostimulation by bacterial DNA and synthetic oligonucleotides, the effects of heat shock protein 90 (Hsp90) inhibitors on the activation of murine spleen cells and macrophages by these molecules were investigated. Murine spleen cells and J774 and RAW264.7 macrophages responded to a CpG-containing oligodeoxynucleotide (CpG ODN) and Escherichia coli DNA by increased production of interleukin 6 (IL-6), IL-12, tumor necrosis factor alpha, and nitric oxide (NO). Pretreatment with any of the three Hsp90 inhibitors geldanamycin, radicicol, and herbimycin A resulted in a dose-dependent suppression of cytokine production from the spleen cells and macrophages and of NO from macrophages stimulated with CpG ODN or E. coli DNA. These Hsp90 inhibitors, however, had no effect on Staphylococcus aureus Cowan strain 1-induced IL-12 production from either the murine spleen cells or macrophages. CpG ODN and E. coli DNA induced increased intracellular levels of phosphorylated extracellular signal-regulated kinases (ERK1 and -2), which are members of the mitogen-activated protein (MAP) kinase family, while geldanamycin and radicicol blocked the phosphorylation of ERK1 and -2 in J774 and RAW264.7 cells. These data indicate that DNA-induced activation of murine spleen cells and macrophages is mediated by Hsp90 and that Hsp90 inhibitor suppression of DNA-induced macrophage activation is associated with disruption of the MAP kinase signaling pathway. Our findings suggest that Hsp90 inhibitors may provide a useful means of elucidating the mechanisms of immunostimulation by bacterial DNA and CpG ODN as well as a strategy for preventing adverse effects of bacterial DNA as well as lipopolysaccharide.

Authors
Zhu, FG; Pisetsky, DS
MLA Citation
Zhu, FG, and Pisetsky, DS. "Role of the heat shock protein 90 in immune response stimulation by bacterial DNA and synthetic oligonucleotides." Infect Immun 69.9 (September 2001): 5546-5552.
PMID
11500428
Source
pubmed
Published In
Infection and immunity
Volume
69
Issue
9
Publish Date
2001
Start Page
5546
End Page
5552

Interleukin-1, tumor necrosis factor alpha, and interleukin-17 synergistically up-regulate nitric oxide and prostaglandin E2 production in explants of human osteoarthritic knee menisci.

OBJECTIVE: In osteoarthritis (OA), a combination of biochemical and biomechanical factors may damage both menisci and articular cartilage. Nitric oxide (NO) and prostaglandin E2 (PGE2) have been implicated as mediators of inflammation in OA. The goals of this study were to determine if menisci from patients with OA produce NO and PGE2, and if the proinflammatory cytokines interleukin-1beta (IL-1beta), tumor necrosis factor a (TNFalpha), and IL-17 augment NO and PGE2 production by these tissues. METHODS: Menisci were obtained from 17 patients (age 47-75 years) undergoing total knee replacement for OA. Tissue explants were cultured alone or with IL-1beta, IL-17, or TNFalpha, and the release of NO and PGE2 from the tissue as well as the presence of type 2 nitric oxide synthase (NOS2) and cyclooxygenase 2 (COX-2) antigens were measured. RESULTS: All menisci constitutively produced NO, and significant increases in NO production were observed in the presence of IL-1beta, TNFalpha, or IL-17 (P < 0.05). The combination of IL-17 and TNFalpha significantly increased NO production compared with either cytokine alone. Basal and cytokine-stimulated NO synthesis was inhibited by the NOS inhibitors NG-monomethyl-L-arginine or N-3-aminoethylbenzylacetamidine (1400W). IL-1beta significantly increased PGE2 production. The combination of IL-1beta and TNFalpha had an additive effect on PGE2 production, while addition of IL-17 to TNFalpha or IL-1beta synergistically enhanced PGE2 production. Inhibition of NO production by 1400W significantly increased IL-1beta-stimulated PGE2 production, and inhibition of PGE2 production by the COX-2 inhibitor N-[2-(cyclohexyloxy)-4-nitrophenyl]-methanesulfonamide significantly increased IL-17-stimulated NO production. CONCLUSION: Menisci from humans with OA spontaneously produced NO and PGE2 in a manner that was synergistically or additively augmented by cytokines. NO and PGE2 exhibited reciprocal regulatory effects on one another, suggesting that pharmaceutical agents designed to inhibit NOS2 or COX-2 production may in fact be influencing both pathways.

Authors
LeGrand, A; Fermor, B; Fink, C; Pisetsky, DS; Weinberg, JB; Vail, TP; Guilak, F
MLA Citation
LeGrand, A, Fermor, B, Fink, C, Pisetsky, DS, Weinberg, JB, Vail, TP, and Guilak, F. "Interleukin-1, tumor necrosis factor alpha, and interleukin-17 synergistically up-regulate nitric oxide and prostaglandin E2 production in explants of human osteoarthritic knee menisci." Arthritis Rheum 44.9 (September 2001): 2078-2083.
PMID
11592370
Source
pubmed
Published In
Arthritis and Rheumatism
Volume
44
Issue
9
Publish Date
2001
Start Page
2078
End Page
2083
DOI
10.1002/1529-0131(200109)44:9<2078::AID-ART358>3.0.CO;2-J

The role of cpg sequences in the induction of anti-DNA antibodies.

To investigate the role of CpG sequences in anti-DNA induction, immunization experiments were performed in mice to assess the immunogenicity of native Escherichia coli (EC) and calf thymus (CT) in incomplete Freund's adjuvant. The effects of CpG sequences were further tested by comparing the adjuvant properties of a synthetic phosphorothioate oligonucleotide with a CpG motif to one with a GpC sequence. Both EC and CT DNA alone induced a limited anti-DNA response. For CT DNA, the addition of a CpG ODN significantly enhanced responses whereas for EC DNA, the presence of a CpG oligonucleotide (ODN) or control GpC ODN did not increase responses compared to EC DNA alone. Specificity analysis by ELISA indicated that these immunizations led to the generation of cross-reactive anti-DNA autoantibodies. These results thus extend the adjuvant effects of CpG sequences to self antigens and suggest mechanisms by which self and foreign antigens can interact in the generation of autoimmunity.

Authors
Pisetsky, DS; Wenk, KS; Reich, CF
MLA Citation
Pisetsky, DS, Wenk, KS, and Reich, CF. "The role of cpg sequences in the induction of anti-DNA antibodies." Clin Immunol 100.2 (August 2001): 157-163.
PMID
11465944
Source
pubmed
Published In
Clinical Immunology
Volume
100
Issue
2
Publish Date
2001
Start Page
157
End Page
163
DOI
10.1006/clim.2001.5064

Mechanical stress and nitric oxide influence leukotriene production in cartilage.

Nitric oxide (NO) and leukotrienes regulate a variety of processes in joint tissues and are frequently elevated in arthritis. Mechanical stress can induce biochemical and functional changes in cartilage that may influence mediator production. To investigate the relationship between mechanical stress and the production of leukotriene B(4) (LTB(4)) and NO, explants of porcine articular cartilage were subjected to mechanical compression for 1 h followed by 23 h recovery in the presence or absence of the NOS2 inhibitor 1400W. Dynamic compression significantly increased LTB(4) and LOX protein production in the presence of 1400W. The induced LTB(4) was functional as evidenced by its ability to promote chemotaxis of RBL-2H3 cells expressing the LTB(4) receptor. Increased LOX protein but not LTB(4) occurred in response to compression alone. These findings provide a direct link between mechanical stress and inflammation in cartilage and may have implications in the pathogenesis and treatment of arthritis.

Authors
Fermor, B; Haribabu, B; Weinberg, JB; Pisetsky, DS; Guilak, F
MLA Citation
Fermor, B, Haribabu, B, Weinberg, JB, Pisetsky, DS, and Guilak, F. "Mechanical stress and nitric oxide influence leukotriene production in cartilage." Biochem Biophys Res Commun 285.3 (July 20, 2001): 806-810.
PMID
11453664
Source
pubmed
Published In
Biochemical and Biophysical Research Communications
Volume
285
Issue
3
Publish Date
2001
Start Page
806
End Page
810
DOI
10.1006/bbrc.2001.5237

The effects of static and intermittent compression on nitric oxide production in articular cartilage explants.

Nitric oxide (NO) production and NO synthase (NOS) expression are increased in osteoarthritis and rheumatoid arthritis, suggesting that NO may play a role in the destruction of articular cartilage. To test the hypothesis that mechanical stress may increase NO production by chondrocytes, we measured the effects of physiological levels of static and intermittent compression on NOS activity, NO production, and NOS antigen expression by porcine articular cartilage explants. Static compression significantly increased NO production at 0.1 MPa stress for 24 h (P < 0.05). Intermittent compression at 0.5 Hz for 6 h followed by 18 h recovery also increased NO production and NOS activity at 1.0 MPa stress (P < 0.05). Intermittent compression at 0.5 Hz for 24 h at a magnitude of 0.1 or 0.5 MPa caused an increase in NO production and NOS activity (P < 0.05). Immunoblot analysis showed stress-induced upregulation of NOS2, but not NOS1 or NOS3. There was no loss in cell viability following any of the loading regimens. Addition of 2 mM 1400 W (a specific NOS2 inhibitor) reduced NO production by 51% with no loss of cell viability. These findings indicate that NO production by chondrocytes is influenced by mechanical compression in vitro and suggest that biomechanical factors may in part regulate NO production in vivo.

Authors
Fermor, B; Weinberg, JB; Pisetsky, DS; Misukonis, MA; Banes, AJ; Guilak, F
MLA Citation
Fermor, B, Weinberg, JB, Pisetsky, DS, Misukonis, MA, Banes, AJ, and Guilak, F. "The effects of static and intermittent compression on nitric oxide production in articular cartilage explants." J Orthop Res 19.4 (July 2001): 729-737.
PMID
11518285
Source
pubmed
Published In
Journal of Orthopaedic Research
Volume
19
Issue
4
Publish Date
2001
Start Page
729
End Page
737
DOI
10.1016/S0736-0266(00)00049-8

The effect of dynamic mechanical compression on nitric oxide production in the meniscus.

OBJECTIVE: The menisci play an important role in the biomechanics of the knee, and loss of meniscal function has been associated with progressive degenerative changes of the joint in rheumatoid arthritis as well as in osteoarthritis. However, little is known about the underlying mechanisms that link meniscal injury or degeneration to arthritis. Meniscal fibrochondrocytes respond to environmental mediators such as growth factors and cytokines, but the influence of mechanical stress on their metabolic activity is not well understood. Nitric oxide (NO) is believed to play a role in mechanical signal transduction, and there is also significant evidence of its role in cartilage and meniscus degeneration. The goal of this study was to determine if meniscal fibrochondrocytes respond to mechanical stress by increasing NO production in vitro. DESIGN: Explants of lateral and medial porcine menisci were dynamically compressed in a precisely controlled manner, and NO production, nitric oxide synthase antigen expression and cell viability were measured. The relative responses of the meniscal surface and deep layers to dynamic compression were also investigated separately. RESULTS: Meniscal NO production was significantly (P< 0.01) increased by dynamic compression in both the medial and lateral menisci. Dynamically compressed menisci contained inducible nitric oxide synthase antigen, while uncompressed menisci did not. Significant (P< 0.05) zonal differences were observed in basal and compression-induced NO production. DISCUSSION: Our findings provide direct evidence that dynamic mechanical stress influences the biological activity of meniscal cells. These results suggest that NO production in vivo may be in part regulated by mechanical stress acting upon the menisci. Since NO affects matrix metabolism in various intraarticular tissues, alterations in the distribution and magnitude of stress in the menisci may have important metabolic as well as biomechanical consequences on joint physiology and function.

Authors
Fink, C; Fermor, B; Weinberg, JB; Pisetsky, DS; Misukonis, MA; Guilak, F
MLA Citation
Fink, C, Fermor, B, Weinberg, JB, Pisetsky, DS, Misukonis, MA, and Guilak, F. "The effect of dynamic mechanical compression on nitric oxide production in the meniscus." Osteoarthritis Cartilage 9.5 (July 2001): 481-487.
PMID
11467897
Source
pubmed
Published In
Osteoarthritis and Cartilage
Volume
9
Issue
5
Publish Date
2001
Start Page
481
End Page
487
DOI
10.1053/joca.2001.0415

The role of the macrophage scavenger receptor in immune stimulation by bacterial DNA and synthetic oligonucleotides.

To assess the role of the macrophage scavenger receptor type A (SRA) in immune activation by CpG DNA, cytokine induction and DNA uptake were tested in vitro and in vivo using SRA knockout (SRA-/-) and wild type (WT) mice. As a source of CpG DNA, Escherichia coli DNA (EC DNA) and a 20-mer phosphorothioate oligodeoxynucleotide with two CpG motifs (CpG ODN) were used. In vitro, both EC DNA and the CpG ODN induced dose-dependent increases of interleukin (IL)-12 production by spleen cells and bone-marrow-derived macrophages (BMMPhi) from both SRA-/- and WT mice. The levels of cytokines produced by SRA-/- spleen cells and BMMPhi were similar to those of WT spleen cells and BMMPhi. When injected intravenously with CpG ODN and EC DNA, both SRA-/- and WT mice showed elevated serum levels of IL-12. To investigate further the role of the SRA, flow cytometry and confocal microscopy were performed to examine the uptake of fluorescently labelled oligonucleotides. SRA-/- and WT BMMPhi showed similarity in the extent of uptake and distribution of oligonucleotides as assessed by these two techniques. Together, these findings indicate that, while the SRA may bind DNA, this receptor is not essential for the uptake of CpG DNA or its immunostimulatory activity.

Authors
Zhu, FG; Reich, CF; Pisetsky, DS
MLA Citation
Zhu, FG, Reich, CF, and Pisetsky, DS. "The role of the macrophage scavenger receptor in immune stimulation by bacterial DNA and synthetic oligonucleotides." Immunology 103.2 (June 2001): 226-234.
PMID
11412310
Source
pubmed
Published In
Immunology
Volume
103
Issue
2
Publish Date
2001
Start Page
226
End Page
234

The effects of intravenous doxycycline therapy for rheumatoid arthritis: a randomized, double-blind, placebo-controlled trial.

OBJECTIVE: To determine the feasibility, safety, and potential clinical efficacy of intravenous (IV) doxycycline therapy for rheumatoid arthritis (RA), as well as its possible effects on serum and urinary markers of collagen breakdown. METHODS: The exploratory trial was designed as a 16-week, single-center, randomized, double-blind, placebo-controlled trial. Eligible subjects with active seropositive or erosive RA were randomly allocated into 3 treatment groups: doxycycline 200 mg IV, azithromycin 250 mg orally, or placebo. The blinded IV study drug was administered once daily for the first 3 weeks by home self-infusion and then weekly for the next 8 weeks, concurrent with the blinded oral study drug at the prescribed doses. The primary end points were the change between baseline and week 4 in the tender joint count, erythrocyte sedimentation rate, and urinary excretion of pyridinoline. RESULTS: The trial was stopped prematurely after enrollment of 31 patients. Three subjects were withdrawn because of worsening arthritis, and 1 patient was withdrawn when newly diagnosed with breast cancer. Infusion-related events occurred in 13 (42%) of 31 patients, but none were serious. There were 4 serious adverse events unrelated to the study drug, including a new diagnosis of breast cancer in 3 cases and hospitalization for abdominal pain in 1 case. No significant differences were observed across treatment groups in any of the 3 primary clinical end points. CONCLUSION: Although IV doxycycline therapy was generally well-tolerated by patients in this trial, it did not show any evidence of reducing disease activity or collagen crosslink production.

Authors
St Clair, EW; Wilkinson, WE; Pisetsky, DS; Sexton, DJ; Drew, R; Kraus, VB; Greenwald, RA
MLA Citation
St Clair, EW, Wilkinson, WE, Pisetsky, DS, Sexton, DJ, Drew, R, Kraus, VB, and Greenwald, RA. "The effects of intravenous doxycycline therapy for rheumatoid arthritis: a randomized, double-blind, placebo-controlled trial." Arthritis Rheum 44.5 (May 2001): 1043-1047.
PMID
11357896
Source
pubmed
Published In
Arthritis and Rheumatism
Volume
44
Issue
5
Publish Date
2001
Start Page
1043
End Page
1047
DOI
10.1002/1529-0131(200105)44:5<1043::AID-ANR183>3.0.CO;2-C

A genomewide screen in multiplex rheumatoid arthritis families suggests genetic overlap with other autoimmune diseases.

Rheumatoid arthritis (RA) is an autoimmune/inflammatory disorder with a complex genetic component. We report the first major genomewide screen of multiplex families with RA gathered in the United States. The North American Rheumatoid Arthritis Consortium, using well-defined clinical criteria, has collected 257 families containing 301 affected sibling pairs with RA. A genome screen for allele sharing was performed, using 379 microsatellite markers. A nonparametric analysis using SIBPAL confirmed linkage of the HLA locus to RA (P < .00005), with lambdaHLA = 1.79. However, the analysis also revealed a number of non-HLA loci on chromosomes 1 (D1S235), 4 (D4S1647), 12 (D12S373), 16 (D16S403), and 17 (D17S1301), with evidence for linkage at a significance level of P<.005. Analysis of X-linked markers using the MLOD method from ASPEX also suggests linkage to the telomeric marker DXS6807. Stratifying the families into white or seropositive subgroups revealed some additional markers that showed improvement in significance over the full data set. Several of the regions that showed evidence for nominal significance (P < .05) in our data set had previously been implicated in RA (D16S516 and D17S1301) or in other diseases of an autoimmune nature, including systemic lupus erythematosus (D1S235), inflammatory bowel disease (D4S1647, D5S1462, and D16S516), multiple sclerosis (D12S1052), and ankylosing spondylitis (D16S516). Therefore, genes in the HLA complex play a major role in RA susceptibility, but several other regions also contribute significantly to overall genetic risk.

Authors
Jawaheer, D; Seldin, MF; Amos, CI; Chen, WV; Shigeta, R; Monteiro, J; Kern, M; Criswell, LA; Albani, S; Nelson, JL; Clegg, DO; Pope, R; Schroeder, HW; Bridges, SL; Pisetsky, DS; Ward, R; Kastner, DL; Wilder, RL; Pincus, T; Callahan, LF; Flemming, D; Wener, MH; Gregersen, PK
MLA Citation
Jawaheer, D, Seldin, MF, Amos, CI, Chen, WV, Shigeta, R, Monteiro, J, Kern, M, Criswell, LA, Albani, S, Nelson, JL, Clegg, DO, Pope, R, Schroeder, HW, Bridges, SL, Pisetsky, DS, Ward, R, Kastner, DL, Wilder, RL, Pincus, T, Callahan, LF, Flemming, D, Wener, MH, and Gregersen, PK. "A genomewide screen in multiplex rheumatoid arthritis families suggests genetic overlap with other autoimmune diseases." Am J Hum Genet 68.4 (April 2001): 927-936.
PMID
11254450
Source
pubmed
Published In
The American Journal of Human Genetics
Volume
68
Issue
4
Publish Date
2001
Start Page
927
End Page
936

Role of tumor necrosis factor α and potential benefit of tumor necrosis factor blockade treatment in systemic lupus erythematosus: Comment on the editorial by Pisetsky [2]

Authors
Aringer, M; Steiner, G; Graninger, W; Smolen, J; Pisetsky, DS
MLA Citation
Aringer, M, Steiner, G, Graninger, W, Smolen, J, and Pisetsky, DS. "Role of tumor necrosis factor α and potential benefit of tumor necrosis factor blockade treatment in systemic lupus erythematosus: Comment on the editorial by Pisetsky [2]." Arthritis and Rheumatism 44.7 (2001): 1721-1722.
PMID
11465728
Source
scival
Published In
Arthritis and Rheumatism
Volume
44
Issue
7
Publish Date
2001
Start Page
1721
End Page
1722
DOI
10.1002/1529-0131(200107)44:7<1721::AID-ART302>3.0.CO;2-J

The American College of Rheumatology Journals: New directions for Y2K + 1

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "The American College of Rheumatology Journals: New directions for Y2K + 1." Arthritis and Rheumatism 44.1 (2001): 1-2.
Source
scival
Published In
Arthritis and Rheumatism
Volume
44
Issue
1
Publish Date
2001
Start Page
1
End Page
2
DOI
10.1002/1529-0131(200101)44:1<1::AID-ANR1>3.0.CO;2-2

Tumor necrosis factor alpha blockers and the induction of anti-DNA autoantibodies.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Tumor necrosis factor alpha blockers and the induction of anti-DNA autoantibodies." Arthritis and rheumatism 43.11 (November 2000): 2381-2382. (Academic Article)
Source
manual
Published In
Arthritis and Rheumatism
Volume
43
Issue
11
Publish Date
2000
Start Page
2381
End Page
2382
DOI
10.1002/1529-0131(200011)43:113.0.CO;2-M

Tumor necrosis factor alpha blockers and the induction of anti-DNA autoantibodies.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Tumor necrosis factor alpha blockers and the induction of anti-DNA autoantibodies." Arthritis Rheum 43.11 (November 2000): 2381-2382.
PMID
11083257
Source
pubmed
Published In
Arthritis and Rheumatism
Volume
43
Issue
11
Publish Date
2000
Start Page
2381
End Page
2382
DOI
10.1002/1529-0131(200011)43:11<2381::AID-ANR1>3.0.CO;2-M

Inhibition of murine macrophage IL-12 production by natural and synthetic DNA.

DNA is a complex macromolecule whose immunological properties vary with sequence and structure. To determine whether DNA can inhibit immune responses, the effects of mammalian DNA and synthetic phosphodiester (Po) and phosphorothioate (Ps) oligonucleotides (ODNs) on IL-12 production were tested using murine macrophages. With bacterial DNA as a stimulant, calf thymus DNA and human placenta DNA blocked IL-12 production by splenic and bone marrow macrophages. A (dG)(30) Po ODN and all single-base Ps 30 mer ODNs were also effective inhibitors. The Ps ODNs also blocked IL-12 production induced by lipopolysaccharide (LPS) and a stimulatory Ps ODN. With the J774 cell line, single-base Ps ODNs inhibited IL-12 production induced by bacterial DNA, LPS, and a stimulatory Ps ODN. Together, these results indicate that DNA has inhibitory properties, suggesting that mammalian DNA could limit immune activation during inflammation and counteract the effects of bacterial DNA.

Authors
Pisetsky, DS; Reich, CF
MLA Citation
Pisetsky, DS, and Reich, CF. "Inhibition of murine macrophage IL-12 production by natural and synthetic DNA." Clin Immunol 96.3 (September 2000): 198-204.
PMID
10964537
Source
pubmed
Published In
Clinical Immunology
Volume
96
Issue
3
Publish Date
2000
Start Page
198
End Page
204
DOI
10.1006/clim.2000.4897

Anti-DNA and autoantibodies.

Systemic lupus erythematosus is a prototypic autoimmune disease characterized by antinuclear antibodies (ANAs), including pathogenic specificities to DNA. As shown by recent research, ANA production is a genetically determined process in which self antigens drive B and T cells that have escaped the normal mechanisms of tolerance. Although antibodies can bind isolated protein or nucleic acid species, the in vivo driving antigens likely exist as complexes that have been released from apoptotic cells. The clinical measurement of ANAs, although valuable in assessing diagnosis and prognosis, must nevertheless be interpreted with caution because ANAs, despite their disease associations, can occur in healthy individuals.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Anti-DNA and autoantibodies." Curr Opin Rheumatol 12.5 (September 2000): 364-368. (Review)
PMID
10990170
Source
pubmed
Published In
Current Opinion in Rheumatology
Volume
12
Issue
5
Publish Date
2000
Start Page
364
End Page
368

The role of cell surface receptors in the activation of human B cells by phosphorothioate oligonucleotides.

Phosphorothioate oligodeoxynucleotides (sODN) containing the CpG motif or TCG repeats induce T cell-independent polyclonal activation of human B cells. To elucidate the mechanism of this response, the role of cell surface receptors was investigated. Sepharose beads coated with stimulatory but not nonstimulatory sODNs induced B cell proliferation comparably with soluble sODNs. The B cell stimulatory activity of Sepharose-bound sODN did not result from free sODN released from the beads since media incubated with coated beads were inactive. Using FITC-labeled sODNs as probes, binding to human B cells could be detected by flow cytometry. Binding was rapid, saturable, initially temperature independent, but with a rapid off-rate. Competition studies indicated that both stimulatory sODNs and minimally stimulatory sODNs bound to the same receptor. By contrast, phosphodiester oligonucleotides with the same nucleotide sequence as sODNs and bacterial DNA inhibited the binding of sODNs to B cells minimally. Charge appeared to contribute to the binding of sODNs to B cells since binding of sODNs was competitively inhibited by negatively charged molecules, including fucoidan, poly I, and polyvinyl sulfate. These data indicate that human B cells bind sODNs by a receptor-mediated mechanism that is necessary but not sufficient for polyclonal activation.

Authors
Liang, H; Reich, CF; Pisetsky, DS; Lipsky, PE
MLA Citation
Liang, H, Reich, CF, Pisetsky, DS, and Lipsky, PE. "The role of cell surface receptors in the activation of human B cells by phosphorothioate oligonucleotides." J Immunol 165.3 (August 1, 2000): 1438-1445.
PMID
10903748
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
165
Issue
3
Publish Date
2000
Start Page
1438
End Page
1445

Stimulation of thymocyte proliferation by phosphorothioate DNA oligonucleotides.

DNA is a complex macromolecule the immunological properties of which depend on short sequence motifs called CpG motifs or immunostimulatory sequences (ISS). These sequences are mitogenic for B cells and can stimulate macrophage cytokine production. While these sequences do not directly activate T cells, they can augment effects of stimulation via the TCR. Furthermore, ISS can affect T cells because of macrophage production of IL-12 and IFN-alpha/beta. In these studies, we further evaluated the immune effects of DNA on T cells, testing the possibility that certain T cell populations can respond directly to this stimulus. We therefore tested the in vitro responses of thymocytes to a series of phosphodiester (Po) and phosphorothioate (Ps) oligonucleotides (ODNs) varying in sequence. In in vitro cultures, phosphorothioate ODNs (sODNs) containing CpG motifs induced significant proliferation of murine thymocytes, although phosphodiester compounds lacked activity. The magnitude of stimulation varied with sequences flanking the CpG motifs, as both dA and dT sequences enhanced the stimulatory capacity of the CpG motif. Furthermore, CpG sODNs were strong costimulators of anti-CD3-mediated thymocyte activation, increasing proliferation compared to anti-CD3 in the absence of DNA. This activation was only partially inhibited by cyclosporine A and was not dependent on a calcium influx. Together, these results indicate that phosphorothioate oligonucleotides containing CpG motifs can directly induce thymocyte proliferation as well as augment TCR activation. These observations thus extend the range of actions of CpG DNA and suggest additional mechanisms for its function as an immunomodulatory agent or adjuvant.

Authors
Mannon, RB; Nataraj, C; Pisetsky, DS
MLA Citation
Mannon, RB, Nataraj, C, and Pisetsky, DS. "Stimulation of thymocyte proliferation by phosphorothioate DNA oligonucleotides." Cell Immunol 201.1 (April 10, 2000): 14-21.
PMID
10805969
Source
pubmed
Published In
Cellular Immunology
Volume
201
Issue
1
Publish Date
2000
Start Page
14
End Page
21
DOI
10.1006/cimm.2000.1635

Tumor necrosis factor blockers in rheumatoid arthritis.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Tumor necrosis factor blockers in rheumatoid arthritis." N Engl J Med 342.11 (March 16, 2000): 810-811.
PMID
10717018
Source
pubmed
Published In
The New England journal of medicine
Volume
342
Issue
11
Publish Date
2000
Start Page
810
End Page
811
DOI
10.1056/NEJM200003163421110

The role of bacterial DNA in autoantibody induction.

Bacterial DNA has potent immunological properties that can stimulate the immune system in SLE in both specific and non-specific ways. As such, this molecule may play an important role in disease pathogenesis, because it can exert immunomodulatory activity and function as a molecular mimic. Future studies will hopefully both determine the role of foreign nucleic acids in the induction of autoantibodies and lead to strategies for their elimination.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "The role of bacterial DNA in autoantibody induction." Curr Top Microbiol Immunol 247 (2000): 143-155. (Review)
PMID
10689785
Source
pubmed
Published In
Current topics in microbiology and immunology
Volume
247
Publish Date
2000
Start Page
143
End Page
155

The antigenic properties of bacterial DNA in normal and aberrant immunity.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "The antigenic properties of bacterial DNA in normal and aberrant immunity." Springer Semin Immunopathol 22.1-2 (2000): 153-166. (Review)
PMID
10944810
Source
pubmed
Published In
Springer seminars in immunopathology
Volume
22
Issue
1-2
Publish Date
2000
Start Page
153
End Page
166

In vitro assay of immunostimulatory activities of plasmid vectors.

DNA vaccination represents a powerful new approach for the elicitation of long-lived protective immunity against a broad range of protein antigens (1,2). In this approach, the vaccine is a plasmid DNA vector that encodes a foreign protein to be targeted for the induction of humoral or cellular responses. Following administration by various routes, the plasmid is taken up by cells to allow intracellular production of the protein for presentation to the immune system. Although the trafficking of the plasmid and its protein product is not well understood, the generation of responses ultimately involves a bone marrow-derived antigen presenting cell (3).

Authors
Reich, CF; Pisetsky, DS
MLA Citation
Reich, CF, and Pisetsky, DS. "In vitro assay of immunostimulatory activities of plasmid vectors." Methods Mol Med 29 (2000): 173-184.
PMID
21374319
Source
pubmed
Published In
Methods in Molecular Medicine
Volume
29
Publish Date
2000
Start Page
173
End Page
184
DOI
10.1385/1-59259-688-6:173

Immune responses to DNA in normal and aberrant immunity.

Because of structural microheterogeneity, DNA can exert powerful effects that lead to immune system activation as well as antibody induction. These activating effects resemble those ofendotoxin and result from sequences that occur much more commonly in bacterial DNA than in mammalian DNA. In contrast, mammalian DNA can inhibit the response to bacterial DNA as well as other stimuli and may serve a counterregulatory role during infection. The recognition of the immune effects of DNA is relevant to the pathogenesis of a variety of infectious and inflammatory diseases including systemic lupus erythematosus, a prototypic autoimmune disease characterized by anti-DNA autoantibodies.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Immune responses to DNA in normal and aberrant immunity." Immunol Res 22.2-3 (2000): 119-126. (Review)
PMID
11339349
Source
pubmed
Published In
Immunologic Research
Volume
22
Issue
2-3
Publish Date
2000
Start Page
119
End Page
126
DOI
10.1385/IR:22:2-3:119

Mechanisms of immune stimulation by bacterial DNA.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Mechanisms of immune stimulation by bacterial DNA." Springer Semin Immunopathol 22.1-2 (2000): 21-33. (Review)
PMID
10944797
Source
pubmed
Published In
Springer seminars in immunopathology
Volume
22
Issue
1-2
Publish Date
2000
Start Page
21
End Page
33

Influence of backbone chemistry on immune activation by synthetic oligonucleotides.

Depending on base sequence, DNA displays immunological activities relevant to the design of novel therapeutic agents. To determine the influence of backbone structure on these activities, we tested a series of synthetic phosphodiester and phosphorothioate oligonucleotides in in vitro cultures of murine spleen cells. These compounds were 30 bases long and consisted of either a single base or an immunostimulatory sequence (AACGTT) flanked on 5' and 3' ends by 12 nucleotides of each base. Cell activation was assessed by both thymidine incorporation and expression of cell surface CD69; production of interleukin-6 and interleukin-12 was used as a measure of cytokine stimulation. In these assays, phosphorothioate oligonucleotides induced much higher levels of proliferation, CD69 expression, and cytokine production than the comparable phosphodiester compounds and had activity at lower concentrations. The sequence for optimal stimulation by phosphorothioates varied among responses, however. For example, whereas compounds containing an immunostimulatory sequence all induced similar levels of proliferation and CD69 expression, cytokine production was greatest with compounds with dA and dT flanks. Furthermore, while single base dG oligonucleotides stimulated proliferation as both phosphodiesters and phosphorothioates, they failed to stimulate cytokine production. Together, these findings indicate that base sequence as well as backbone chemistry influence immune activation by synthetic oligonucleotides, with the effects varying among responses. While suggesting differences in the structure-function relationships of nucleic acids in their immune activities, these findings also raise the possibility of the design of agents with specific patterns of immune modulation.

Authors
Pisetsky, DS; Reich, CF
MLA Citation
Pisetsky, DS, and Reich, CF. "Influence of backbone chemistry on immune activation by synthetic oligonucleotides." Biochem Pharmacol 58.12 (December 15, 1999): 1981-1988.
PMID
10591154
Source
pubmed
Published In
Biochemical Pharmacology
Volume
58
Issue
12
Publish Date
1999
Start Page
1981
End Page
1988

Multivalent cross-linking of membrane Ig sensitizes murine B cells to a broader spectrum of CpG-containing oligodeoxynucleotide motifs, including their methylated counterparts, for stimulation of proliferation and Ig secretion.

We have previously reported that B cells that are activated by multivalent but not bivalent membrane Ig cross-linking ligands synergize with various B cell activators culminating in enhanced B cell proliferation. In this study we asked whether B cells that are activated by a multivalent mIg cross-linking agonist could respond to oligodeoxynucleotides (ODN) containing non-stimulatory motifs. Earlier reports have shown that ODN containing a CpG motif in which the cytosine is unmethylated and is flanked by two 5' purines and two 3' pyrimidines induce high levels of B cell activation, while ODN whose CpG are methylated or flanked by sequences other than the optimal two 5' purines and two 3' pyrimidines were non-stimulatory. In this manuscript we show that when B cells are stimulated in vitro with dextran-conjugated anti-IgD antibodies (anti-IgD-dex), as the multivalent mIg ligand, their proliferation is enhanced and they can be induced to secrete Ig in response to ODN containing various non-optimal motifs, both methylated and non-methylated. Furthermore we could induce synergistic levels of proliferation with concentrations of anti-IgD-dex that were in the picomolar concentration range and with concentrations of ODN that were 10- to 100-fold less than previously reported to be necessary for mitogenic activity. These data provided a model to explain how low concentrations of a multi-epitope-expressing microorganism in the context of mammalian (methylated) or microorganism (non-methylated) DNA can lead to dysregulated B cell proliferation and Ig secretion.

Authors
Goeckeritz, BE; Flora, M; Witherspoon, K; Vos, Q; Lees, A; Dennis, GJ; Pisetsky, DS; Klinman, DM; Snapper, CM; Mond, JJ
MLA Citation
Goeckeritz, BE, Flora, M, Witherspoon, K, Vos, Q, Lees, A, Dennis, GJ, Pisetsky, DS, Klinman, DM, Snapper, CM, and Mond, JJ. "Multivalent cross-linking of membrane Ig sensitizes murine B cells to a broader spectrum of CpG-containing oligodeoxynucleotide motifs, including their methylated counterparts, for stimulation of proliferation and Ig secretion." Int Immunol 11.10 (October 1999): 1693-1700.
PMID
10508187
Source
pubmed
Published In
International Immunology
Volume
11
Issue
10
Publish Date
1999
Start Page
1693
End Page
1700

The influence of lipofectin on the in vitro stimulation of murine spleen cells by bacterial DNA and plasmid DNA vectors.

Lipofectin is a mixture of two cationic lipids, N-[1-(2,3-dioleyloxy)propyl]-n,n,n-trimethylammonium chloride (DOTMA) and dioleoyl phosphotidylethanolamine (DOPE), and has been commonly used to promote transfection of plasmid vectors in vitro and in vivo. In these experiments, the effect of lipofectin on in vitro immunostimulation by bacterial and plasmid DNA was tested to determine if these lipids can also influence immune properties of DNA. As a model, spleen cells from BALB/c and C3H/HeJ mice were cultured with DNA from either Escherichia coli DNA or the pEGFP-N1 plasmid at various ratios with lipofectin. As an index of immune stimulation, in vitro proliferation as well as production of interleukin-12 (IL-12) and interferon-gamma (IFN-gamma) were assessed. For both bacterial DNA and plasmid DNA, the presence of lipofectin led to a marked increase in the production of IFN-gamma under conditions in which increases in IL-12 production were limited. The IFN-gamma production was nevertheless dependent on IL-12, as shown by the effects of anti-IL-12 antibodies. Under these culture conditions, lipofectin did not significantly augment proliferation induced by DNA. These findings indicate that lipofectin can increase the in vitro immunostimulatory effects of bacterial and plasmid DNA, although the magnitude of the increase may vary among responses.

Authors
Pisetsky, DS; Reich, CF
MLA Citation
Pisetsky, DS, and Reich, CF. "The influence of lipofectin on the in vitro stimulation of murine spleen cells by bacterial DNA and plasmid DNA vectors." J Interferon Cytokine Res 19.10 (October 1999): 1219-1226.
PMID
10547162
Source
pubmed
Published In
Journal of Interferon & Cytokine Research
Volume
19
Issue
10
Publish Date
1999
Start Page
1219
End Page
1226
DOI
10.1089/107999099313163

A piece of my mind. What we are made of.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "A piece of my mind. What we are made of." JAMA 282.12 (September 22, 1999): 1112-.
PMID
10501098
Source
pubmed
Published In
JAMA : the journal of the American Medical Association
Volume
282
Issue
12
Publish Date
1999
Start Page
1112

Deficiency of 5-lipoxygenase abolishes sex-related survival differences in MRL-lpr/lpr mice.

Leukotrienes, the 5-lipoxygenase (5LO) products of arachidonic acid metabolism, have many proinflammatory actions that have been implicated in the pathogenesis of a variety of inflammatory diseases. To investigate the role of LTs in autoimmune disease, we generated an MRL-lpr/lpr mouse line with a targeted disruption of the 5lo gene. MRL-lpr/lpr mice spontaneously develop autoimmune disease that has many features resembling human systemic lupus erythematosus, including sex-related survival differences; female MRL-lpr/lpr mice experience significant early mortality compared with males. Unexpectedly, we found that mortality was accelerated in male 5LO-deficient MRL-lpr/lpr mice compared with male wild-type MRL-lpr/lpr animals. In contrast, the 5lo mutation had no effect on survival in females. Mortality was also accelerated in male MRL-lpr/lpr mice that were treated chronically with a pharmacological inhibitor of LT synthesis. Furthermore, LT-dependent inflammatory responses are enhanced in male MRL-lpr/lpr mice compared with females, and the 5lo mutation has greater impact on these responses in males. Because immune complex-mediated glomerulonephritis is the major cause of death in MRL-lpr/lpr mice and has been related to arachidonic acid metabolites, we also assessed kidney function and histopathology. In male MRL-lpr/lpr mice, renal plasma flow was significantly reduced in the 5lo-/- compared with the 5lo+/+ group, although there were no differences in the severity of renal histopathology, lymphoid hyperplasia, or arthritis between the groups. These findings suggest that the presence of a functional 5lo gene confers a survival advantage on male MRL-lpr/lpr mice and that, when 5LO function is inhibited, either genetically or pharmacologically, this advantage is abolished.

Authors
Goulet, JL; Griffiths, RC; Ruiz, P; Spurney, RF; Pisetsky, DS; Koller, BH; Coffman, TM
MLA Citation
Goulet, JL, Griffiths, RC, Ruiz, P, Spurney, RF, Pisetsky, DS, Koller, BH, and Coffman, TM. "Deficiency of 5-lipoxygenase abolishes sex-related survival differences in MRL-lpr/lpr mice." J Immunol 163.1 (July 1, 1999): 359-366.
PMID
10384136
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
163
Issue
1
Publish Date
1999
Start Page
359
End Page
366

The influence of DNA size on the binding of antibodies to DNA in the sera of normal human subjects and patients with systemic lupus erythematosus (SLE).

To elucidate antibody recognition of DNA in normal and aberrant immunity, the binding of sera of normal human subjects (NHS) and patients with SLE was tested with mammalian and bacterial DNA varying in size. Klebsiella pneumoniae (KP) and calf thymus (CT) single-stranded DNA (ssDNA) were investigated as model antigens using the restriction enzyme HinfI to generate fragments with the size range of 800-5000 base pairs. The influence of size on activity was assessed by ELISA by both titration of serum as well as coating antigen concentration. In both assay formats, SLE sera bound equivalently to intact CT and KP DNA, but had dramatically reduced reactivity to fragments of both antigens. In contrast, NHS bound similarly to intact KP DNA and its fragments but had low reactivity to CT DNA. These results suggest that SLE and NHS anti-DNA react with different antigenic determinants on DNA, as shown by cross-reactivity as well as size dependency in solid-phase assays.

Authors
Pisetsky, DS; Gonzalez, TC
MLA Citation
Pisetsky, DS, and Gonzalez, TC. "The influence of DNA size on the binding of antibodies to DNA in the sera of normal human subjects and patients with systemic lupus erythematosus (SLE)." Clin Exp Immunol 116.2 (May 1999): 354-359.
PMID
10337030
Source
pubmed
Published In
Clinical & Experimental Immunology
Volume
116
Issue
2
Publish Date
1999
Start Page
354
End Page
359

A college for its teachers.

Authors
Pisetsky, DS; White, B
MLA Citation
Pisetsky, DS, and White, B. "A college for its teachers." Arthritis and rheumatism 42.4 (April 1999): 595-598. (Academic Article)
Source
manual
Published In
Arthritis and Rheumatism
Volume
42
Issue
4
Publish Date
1999
Start Page
595
End Page
598
DOI
10.1002/1529-0131(199904)42:43.0.CO;2-U

A college for its teachers.

Authors
Pisetsky, DS; White, B
MLA Citation
Pisetsky, DS, and White, B. "A college for its teachers." Arthritis Rheum 42.4 (April 1999): 595-598.
PMID
10211872
Source
pubmed
Published In
Arthritis and Rheumatism
Volume
42
Issue
4
Publish Date
1999
Start Page
595
End Page
598
DOI
10.1002/1529-0131(199904)42:4<595::AID-ANR1>3.0.CO;2-U

Pain in the rheumatic diseases. Practical aspects of diagnosis and treatment.

Patients with rheumatic disease experience pain that can be intense, persistent, and disabling. This pain is frequently multifactorial in origin and has both central and peripheral components. Because of the array of conditions that can cause musculoskeletal pain, patient management must begin with a complete clinical assessment that identifies possible etiologies and measures objective findings against subjective complaints. Especially in patients with known rheumatic disease, the possibility of concurrent pain of central origin must be considered and appropriate treatment given. By applying a comprehensive therapy plan of drugs, physical therapy, and patient education, significant benefits can often be achieved in this prevalent group of painful diseases.

Authors
Rice, JR; Pisetsky, DS
MLA Citation
Rice, JR, and Pisetsky, DS. "Pain in the rheumatic diseases. Practical aspects of diagnosis and treatment." Rheum Dis Clin North Am 25.1 (February 1999): 15-30. (Review)
PMID
10083957
Source
pubmed
Published In
Rheumatic Disease Clinics of North America
Volume
25
Issue
1
Publish Date
1999
Start Page
15
End Page
30

Immunostimulatory properties of genomic DNA from different bacterial species.

Bacterial DNA has potent immunological properties because of its content of immunostimulatory sequences centering on CpG motifs. To investigate whether DNA from various bacterial species differ in these properties, the activity of a panel of DNA was assessed in in vitro cultures of murine spleen cells. This panel varied in base composition and included DNA from Clostridium perfringens (CP), Escherichia coli (EC), Micrococcus lysodeikticus (MC), Staphylococcus aureus (SA), and, as a mammalian DNA control, calf thymus (CT) DNA. In assays of IL-12 and IFN-gamma production as well as B cell mitogenesis, these DNA showed marked differences in their immunostimulatory activity. For both cytokine and B cell responses, EC DNA demonstrated the highest activity while CP DNA had the lowest activity among the bacterial DNA. To determine whether differences in stimulatory capacity resulted from differences in cell uptake, the activity of DNA complexed with lipofectin was tested. While the addition of lipofectin to DNA increased stimulation by all DNA, it did not change the relative potency of the DNA tested. These results indicate that bacterial DNA differ in their immunostimulatory capacity, most likely reflecting their content of CpG motifs. These differences could affect the induction of innate immunity as well as the consequences of infection.

Authors
Neujahr, DC; Reich, CF; Pisetsky, DS
MLA Citation
Neujahr, DC, Reich, CF, and Pisetsky, DS. "Immunostimulatory properties of genomic DNA from different bacterial species." Immunobiology 200.1 (February 1999): 106-119.
PMID
10084699
Source
pubmed
Published In
Immunobiology
Volume
200
Issue
1
Publish Date
1999
Start Page
106
End Page
119
DOI
10.1016/S0171-2985(99)80036-9

The influence of base sequence on the immunostimulatory properties of DNA.

DNA is a complex macromolecule whose immunological properties vary with base sequences. As shown with synthetic oligonucleotides, potent immune stimulation results from six base motifs called CpG motifs or immunostimulatory sequences (ISS). These sequences center on an unmethylated CpG dinucleotide and occur much more commonly in bacterial DNA than mammalian DNA. As such, CpG motifs may function as a danger signal to stimulate B cell activation and cytokine production. In addition to CpG motifs, runs of deoxyguanosine (dG) residues in DNA can induce B cell activation and promote macrophage cytokine expression by adjacent CpG motifs. The array of these sequences may determine the overall immune activity of a DNA molecule and affect such processes as host defense against infection as well as the use of plasmids and synthetic oligonucleotides to treat disease.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "The influence of base sequence on the immunostimulatory properties of DNA." Immunol Res 19.1 (1999): 35-46. (Review)
PMID
10374694
Source
pubmed
Published In
Immunologic Research
Volume
19
Issue
1
Publish Date
1999
Start Page
35
End Page
46
DOI
10.1007/BF02786475

Doing everything [6]

Authors
Knudsen, NS; Pisetsky, DS
MLA Citation
Knudsen, NS, and Pisetsky, DS. "Doing everything [6]." Annals of Internal Medicine 130.2 (1999): 165-166.
PMID
10068376
Source
scival
Published In
Annals of internal medicine
Volume
130
Issue
2
Publish Date
1999
Start Page
165
End Page
166

Specificity of antibodies to bacterial DNA in the sera of healthy human subjects and patients with systemic lupus erythematosus

Objective. To elucidate the epitope structure to DNA by identifying antigenic determinants on bacterial DNA bound by anti-DNA antibodies from normal human subjects (NHS) and patients with systemic lupus erythematosus (SLE). Methods. Sera from NHS and patients with SLE were tested by ELISA for the presence of antibodies to single stranded DNA from calf thymus, Micrococcus lysodeikticus, Staphylococcus epidermidis, Clostridium perfringens, and Klebsiella pneumoniae. To assess binding to conserved and nonconserved determinants, sera were absorbed on DNA-cellulose affinity columns bearing each of the bacterial DNA and then tested for binding to the other DNA antigens. Results. Absorption of SLE sera with any of the bacterial DNA caused a loss of binding to all other bacterial DNA as well as calf thymus DNA. In contrast, absorption of NHS sera with bacterial DNA caused a loss of binding to the DNA on the affinity column with much less effect on binding to the other DNA antigens. Conclusion. These results indicate a marked difference in the specificity of antibodies to bacterial DNA in NHS and patients with SLE. The binding of SLE anti-DNA to predominantly conserved determinants suggests that a shift in patterns of anti-DNA specificity may be associated with the autoimmune state.

Authors
Pisetsky, D; Drayton, D; Wu, Z-Q
MLA Citation
Pisetsky, D, Drayton, D, and Wu, Z-Q. "Specificity of antibodies to bacterial DNA in the sera of healthy human subjects and patients with systemic lupus erythematosus." Journal of Rheumatology 26.9 (1999): 1934-1938.
PMID
10493672
Source
scival
Published In
The Journal of rheumatology
Volume
26
Issue
9
Publish Date
1999
Start Page
1934
End Page
1938

Preface

Authors
Pisetsky, DS; Bradley, LA
MLA Citation
Pisetsky, DS, and Bradley, LA. "Preface." Rheumatic Disease Clinics of North America 25.1 (1999): XI-XII.
Source
scival
Published In
Rheumatic Disease Clinics of North America
Volume
25
Issue
1
Publish Date
1999
Start Page
XI
End Page
XII
DOI
10.1016/S0889-857X(05)70050-8

The binding of anti-DNA antibodies to phosphorothioate oligonucleotides in a solid phase immunoassay.

Phosphorothioate oligonucleotides (S-oligos) are nucleic acid derivatives that are commonly used as antisense agents. These compounds, similar to bacterial DNA and CpG oligonucleotides, display a variety of immunological activities in vitro and in vivo. To assess further these activities, the antigenicity of a series of S-oligos was assessed in the solid phase using anti-DNA antibodies from sera of patients with systemic lupus erythematosus. By ELISA, S-oligos bound well to anti-DNA antibodies under the same conditions as calf thymus DNA antigen. The specificity for anti-DNA was established by competition assays showing cross-inhibition of binding by DNA and S-oligos. Reactivity with anti-DNA was observed with S-oligos varying in sequence, suggesting interaction with a conserved determinant not strictly dependent on the bases. Furthermore. in comparison with a phosphodiester oligomer of the same sequence, a phosphorothioate showed dramatically increased activity. These findings indicate that structural features associated with the S-oligo backbone promote specific binding to anti-DNA antibodies and influence the size requirement for antigenicity in the solid phase. These observations thus extend the immunological properties of S-oligos and suggest uses of these compounds in the diagnosis and treatment of autoimmune disease.

Authors
Pisetsky, DS; Reich, CF
MLA Citation
Pisetsky, DS, and Reich, CF. "The binding of anti-DNA antibodies to phosphorothioate oligonucleotides in a solid phase immunoassay." Mol Immunol 35.18 (December 1998): 1161-1170.
PMID
10199390
Source
pubmed
Published In
Molecular Immunology
Volume
35
Issue
18
Publish Date
1998
Start Page
1161
End Page
1170

The influence of base sequence on the immunological properties of defined oligonucleotides.

To assess the influence of base sequence on the immunostimulatory activities of DNA, cell binding and mitogenicity of a series of 30-mer phosphodiester oligonucleotides were tested using murine spleen cells. These compounds consisted of either a single base or a six base CpG motif in the context of 5' and 3' flanking sequences of each base. Among fluoresceinated oligonucleotides, (dG)30 had the highest binding of single base compounds tested while the presence of dG flanks increased binding of compounds with six base motifs, whether active on inactive. In assays of mitogenesis including incorporation of thymidine and uridine as well as expression of cell surface CD69, (dG)30 induced the highest responses among single base compounds. Among compounds with an active six base motif, the extent of proliferation varied with flanking sequence, with dG flanks producing the greatest stimulation in all assays tested. Together, these findings indicate that a variety of base sequences may affect the immunomodulatory properties of DNA, with the activity of dG sequences perhaps resulting from the formation of variant DNA structures.

Authors
Pisetsky, DS; Reich, CF
MLA Citation
Pisetsky, DS, and Reich, CF. "The influence of base sequence on the immunological properties of defined oligonucleotides." Immunopharmacology 40.3 (November 1998): 199-208.
PMID
9858063
Source
pubmed
Published In
Immunopharmacology
Volume
40
Issue
3
Publish Date
1998
Start Page
199
End Page
208

Effects of bacterial DNA on cytokine production by (NZB/NZW)F1 mice.

Microbial DNA has multiple immune effects including the capacity to induce polyclonal B cell activation and cytokine production in normal mice. We recently described the accelerated induction of anti-DNA Abs in NZB/NZW mice immunized with Escherichia coli (EC) dsDNA; paradoxically these mice developed less renal disease than unimmunized mice or mice immunized with calf thymus DNA. We postulated that alterations in cytokine production induced by bacterial DNA may play a key role in renal protection. To determine the effect of bacterial DNA on cytokine production in NZB/NZW mice, we measured the serum cytokine levels, cell culture supernatant cytokine levels, and number of cytokine-producing splenocytes in NZB/NZW mice injected with EC DNA, calf thymus DNA, or an immune active oligonucleotide. There was a 10- to 25-fold increase in the number of cells secreting IFN-gamma compared with IL-4 in mice immunized with EC DNA. IL-12-secreting cells were also increased by bacterial DNA immunization. In parallel with the increase in IFN-gamma secreting cells, there was a significant rise in serum IFN-gamma levels in mice receiving EC DNA. These results indicate that EC DNA modulates systemic cytokine levels in NZB/NZW mice, selectively increasing IL-12 and IFN-gamma while decreasing IL-4 production. The cytokine response of NZB/NZW mice to bacterial DNA may be of significance in disease pathogenesis and relevant to the treatment of lupus-like disease.

Authors
Gilkeson, GS; Conover, J; Halpern, M; Pisetsky, DS; Feagin, A; Klinman, DM
MLA Citation
Gilkeson, GS, Conover, J, Halpern, M, Pisetsky, DS, Feagin, A, and Klinman, DM. "Effects of bacterial DNA on cytokine production by (NZB/NZW)F1 mice." J Immunol 161.8 (October 15, 1998): 3890-3895.
PMID
9780154
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
161
Issue
8
Publish Date
1998
Start Page
3890
End Page
3895

The influence of DNA sequence on the immunostimulatory properties of plasmid DNA vectors.

To determine the influence of DNA sequence on immunostimulatory properties of vaccine vectors, we tested the induction of in vitro and in vivo immune responses by plasmids modified to contain extended runs of dG sequences. Studies with oligonucleotides indicate that dG sequences can directly stimulate B cells as well as enhance the activity of immunostimulatory CpG motifs because of interaction with the macrophage scavenger receptor (MSR); this receptor can bind a variety of polyanions including dG sequences. To modify vectors, we introduced stretches of 20-60 dG residues into the pCMV-beta and pSG5rab.gp vectors and measured the ability of these plasmids to induce IL-12 and IFN-gamma production by murine splenocytes. The induction of in vivo antibody responses to rabies glycoprotein was also assessed with the pSG5rab.gp vectors. In in vitro cultures, cytokine production induced by plasmids with and without dG sequences was similar. Furthermore, the addition of dG sequences to pSG5rab.gp vectors failed to enhance the anti-rabies glycoprotein response to immunization. To assess further mechanisms by which plasmids stimulate macrophages, we measured the effects of MSR ligands on in vitro cytokine induction. In in vitro cultures, poly(G), dG30, and fucoidan inhibited IL-12 induction by plasmids. IL-12 induction was also inhibited by mammalian DNA but was unaffected by polyanions that are not MSR ligands. Together, these results suggest that the addition of 20 to 60-base dG sequences to plasmids does not significantly affect their properties as immunostimulators or vaccines. Furthermore, these results suggest that MSR ligands can block cytokine induction by plasmid DNA whether or not the plasmid contains extended runs of dG.

Authors
Wloch, MK; Pasquini, S; Ertl, HC; Pisetsky, DS
MLA Citation
Wloch, MK, Pasquini, S, Ertl, HC, and Pisetsky, DS. "The influence of DNA sequence on the immunostimulatory properties of plasmid DNA vectors." Hum Gene Ther 9.10 (July 1, 1998): 1439-1447.
PMID
9681415
Source
pubmed
Published In
Human Gene Therapy
Volume
9
Issue
10
Publish Date
1998
Start Page
1439
End Page
1447
DOI
10.1089/hum.1998.9.10-1439

Doing everything.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Doing everything." Ann Intern Med 128.10 (May 15, 1998): 869-870.
PMID
9599202
Source
pubmed
Published In
Annals of internal medicine
Volume
128
Issue
10
Publish Date
1998
Start Page
869
End Page
870

Anti-DNA antibodies in systemic lupus erythematosus: a case of mistaken identity?

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Anti-DNA antibodies in systemic lupus erythematosus: a case of mistaken identity?." J Rheumatol 25.2 (February 1998): 195-197.
PMID
9489804
Source
pubmed
Published In
The Journal of rheumatology
Volume
25
Issue
2
Publish Date
1998
Start Page
195
End Page
197

Antibody responses to DNA in normal immunity and aberrant immunity.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Antibody responses to DNA in normal immunity and aberrant immunity." Clin Diagn Lab Immunol 5.1 (January 1998): 1-6. (Review)
PMID
9455870
Source
pubmed
Published In
Clinical and diagnostic laboratory immunology
Volume
5
Issue
1
Publish Date
1998
Start Page
1
End Page
6

Immune recognition of DNA in SLE.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Immune recognition of DNA in SLE." Clin Immunol Immunopathol 86.1 (January 1998): 1-2.
PMID
9434790
Source
pubmed
Published In
Clinical Immunology and Immunopathology
Volume
86
Issue
1
Publish Date
1998
Start Page
1
End Page
2

New treatments for rheumatoid arthritis.

Authors
Pisetsky, D
MLA Citation
Pisetsky, D. "New treatments for rheumatoid arthritis." Health news (Waltham, Mass.) 4.14 (1998): 3--.
PMID
9825708
Source
scival
Published In
Health news (Waltham, Mass.)
Volume
4
Issue
14
Publish Date
1998
Start Page
3-

The influence of susceptibility factors on the immune response to DNA.

Susceptibility to autoimmune disease results from genetic factors that determine the pattern of immune responsiveness to self as well as foreign antigens. These factors may influence the immune response to DNA, a complex macromolecule that can induce antibody responses in normal as well as aberrant immunity. In systemic lupus erythematosus, anti-DNA antibodies target conserved sites present on all DNA and appear to arise by a T dependent mechanism. In contrast, in normal humans, anti-DNA antibodies react to non-conserved sites on certain bacterial DNA and have features suggesting induction by a T independent mechanism. The activity of bacterial DNA reflects the presence of base sequence motifs centering on an unmethylated CpG core. Because of susceptibility factors in patients with systemic lupus erythematosus, bacterial DNA may drive a response crossreactive with self DNA instead of a response specific for the foreign antigen.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "The influence of susceptibility factors on the immune response to DNA." Environ Toxicol Pharmacol 4.3-4 (December 1997): 295-298.
PMID
21781836
Source
pubmed
Published In
Environmental Toxicology and Pharmacology
Volume
4
Issue
3-4
Publish Date
1997
Start Page
295
End Page
298

Till death do us part.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Till death do us part." J Gen Intern Med 12.11 (November 1997): 705-706.
PMID
17764022
Source
pubmed
Published In
Journal of General Internal Medicine
Volume
12
Issue
11
Publish Date
1997
Start Page
705
End Page
706
DOI
10.1046/j.1525-1497.1997.07144.x

Clinical and serologic manifestations of autoimmune disease in MRL-lpr/lpr mice lacking nitric oxide synthase type 2.

Nitric oxide (NO) is an important mediator of the inflammatory response. MRL-lpr/lpr mice overexpress inducible nitric oxide synthase (NOS2) and overproduce NO in parallel with the development of an autoimmune syndrome with a variety of inflammatory manifestations. In previous studies, we showed that inhibiting NO production with the nonselective nitric oxide synthase (NOS) inhibitor NG-monomethyl-arginine reduced glomerulonephritis, arthritis, and vasculitis in MRL-lpr/lpr mice. To define further the role of NO and NOS2 in disease in MRL-lpr/lpr mice, mice with targeted disruption of NOS2 were produced by homologous recombination and bred to MRL-lpr/lpr mice to the N4 generation. MRL-lpr/lpr littermates homozygous for disrupted NOS2 (-/-), heterozygous for disrupted NOS2 (+/-), or wildtype (+/+) were derived for this study. Measures of NO production were markedly decreased in the MRL-lpr/lpr (-/-) mice compared with MRL-lpr/lpr (+/+) mice, with intermediate production by the MRL-lpr/lpr (+/-) mice. There was no detectable NOS2 protein by immunoblot analysis of the spleen, liver, kidney, and peritoneal macrophages of the (-/-) animals, whereas that of (+/+) was high and (+/-) intermediate. The (-/-) mice developed glomerular and synovial pathology similar to that of the (+/-) and (+/+) mice. However, (-/-) mice and (+/-) mice had significantly less vasculitis of medium-sized renal vessels than (+/+) mice. IgG rheumatoid factor levels were significantly lower in the (-/-) mice as compared with (+/+) mice, but levels of anti-DNA antibodies were comparable in all groups. Our findings show that NO derived from NOS2 has a variable impact on disease manifestations in MRL-lpr/lpr mice, suggesting heterogeneity in disease mechanisms.

Authors
Gilkeson, GS; Mudgett, JS; Seldin, MF; Ruiz, P; Alexander, AA; Misukonis, MA; Pisetsky, DS; Weinberg, JB
MLA Citation
Gilkeson, GS, Mudgett, JS, Seldin, MF, Ruiz, P, Alexander, AA, Misukonis, MA, Pisetsky, DS, and Weinberg, JB. "Clinical and serologic manifestations of autoimmune disease in MRL-lpr/lpr mice lacking nitric oxide synthase type 2." J Exp Med 186.3 (August 4, 1997): 365-373.
PMID
9236188
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
186
Issue
3
Publish Date
1997
Start Page
365
End Page
373

Immunostimulatory DNA: a clear and present danger?

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Immunostimulatory DNA: a clear and present danger?." Nat Med 3.8 (August 1997): 829-831.
PMID
9256266
Source
pubmed
Published In
Nature Medicine
Volume
3
Issue
8
Publish Date
1997
Start Page
829
End Page
831

Specificity and immunochemical properties of antibodies to bacterial DNA in sera of normal human subjects and patients with systemic lupus erythematosus (SLE).

To elucidate the mechanisms of anti-DNA production, we assessed the binding of sera of normal human subjects (NHS) and patients with SLE to a panel of bacterial and mammalian DNA. Using single-stranded DNA as antigens in an ELISA, NHS showed significant binding to some but not all bacterial DNA, while lacking reactivity to calf thymus DNA. Among bacterial DNA, the highest levels of binding were observed with DNA from Micrococcus lysodeikticus and Staphylococcus aureus. In contrast, SLE sera showed high levels of binding to all DNA tested. To evaluate further immunochemical properties of the anti-DNA antibodies, the subclass distribution of these responses was evaluated by subclass-specific reagents. While NHS showed a predominance of IgG2 antibodies to bacterial DNA, SLE sera had a predominance of IgG1 antibodies to these antigens. Together, these results provide further evidence for the antigenicity of bacterial DNA and suggest that NHS and SLE anti-DNA differ in the patterns of epitope recognition as well as mechanisms of induction.

Authors
Wu, ZQ; Drayton, D; Pisetsky, DS
MLA Citation
Wu, ZQ, Drayton, D, and Pisetsky, DS. "Specificity and immunochemical properties of antibodies to bacterial DNA in sera of normal human subjects and patients with systemic lupus erythematosus (SLE)." Clin Exp Immunol 109.1 (July 1997): 27-31.
PMID
9218820
Source
pubmed
Published In
Clinical & Experimental Immunology
Volume
109
Issue
1
Publish Date
1997
Start Page
27
End Page
31

Molecular properties of anti-DNA induced in preautoimmune NZB/W mice by immunization with bacterial DNA.

To elucidate the mechanism of Ag drive in the anti-DNA response, the Ab response to bacterial DNA has been analyzed in normal and autoimmune mice. Preautoimmune NZB/W mice immunized with Escherichia coli dsDNA produce Abs that resemble spontaneous autoantibodies and bind mammalian dsDNA. In contrast, normal mice, when immunized similarly, produce Abs that bind only bacterial dsDNA. To characterize further the responsiveness of NZB/W mice to bacterial DNA, we determined the molecular properties of mAbs from preautoimmune NZB/W mice immunized with E. coli DNA. Of nine Abs studied, all were IgM and all bound mammalian ssDNA, while four had appreciable reactivity with mammalian dsDNA. The induced anti-dsDNA resembled spontaneous anti-DNA from autoimmune mice in V gene utilization and V(H) CDR3 arginine content. These Abs lacked evidence of somatic mutation, however, indicating that affinity maturation via somatic mutation is not essential for dsDNA reactivity. The findings suggest that preautoimmune NZB/W mice have immunoregulatory defects that allow activation of mammalian dsDNA reactive B cells by bacterial DNA.

Authors
Wloch, MK; Alexander, AL; Pippen, AM; Pisetsky, DS; Gilkeson, GS
MLA Citation
Wloch, MK, Alexander, AL, Pippen, AM, Pisetsky, DS, and Gilkeson, GS. "Molecular properties of anti-DNA induced in preautoimmune NZB/W mice by immunization with bacterial DNA." J Immunol 158.9 (May 1, 1997): 4500-4506.
PMID
9127017
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
158
Issue
9
Publish Date
1997
Start Page
4500
End Page
4506

Deficient expression of antibodies specific for bacterial DNA by patients with systemic lupus erythematosus.

Antibodies to DNA occur commonly in the sera of normal human subjects and bind to nonconserved sites exclusive to DNA from certain bacterial species. These antibodies are primarily IgG2 and differ from anti-DNA antibodies in systemic lupus erythematosus (SLE) sera, which are primarily IgG1 and bind conserved sites on all DNA. To investigate the immune response to bacterial DNA in SLE, antibodies to single-stranded DNA from Micrococcus lysodeikticus (MC) were measured in normal human subjects and in SLE sera by enzyme-linked immunosorbent assay either directly or following absorption by cellulose bearing calf thymus (CT) DNA; isotype-specific reagents were used to assess the IgG2 response. In these assays, SLE sera had a marked deficiency in antibodies specific for MC single-stranded DNA as demonstrated by the reduction of antibody levels by CT DNA cellulose absorption. Furthermore, IgG2 anti-DNA in SLE sera were cross-reactive with MC and CT single-stranded DNA, whereas IgG2 anti-DNA in normal human subjects' sera were specific for the bacterial DNA antigen. Together, these findings indicate that SLE patients have a marked deficiency in the production of antibodies specific for bacterial DNA antigen. This deficiency could predispose to anti-DNA autoantibody production and result from tolerance defects that distort the array of specificities in the B-cell repertoire.

Authors
Pisetsky, DS; Drayton, DM
MLA Citation
Pisetsky, DS, and Drayton, DM. "Deficient expression of antibodies specific for bacterial DNA by patients with systemic lupus erythematosus." Proc Assoc Am Physicians 109.3 (May 1997): 237-244.
PMID
9154640
Source
pubmed
Published In
Proceedings of the Association of American Physicians
Volume
109
Issue
3
Publish Date
1997
Start Page
237
End Page
244

DNA and the immune system.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "DNA and the immune system." Ann Intern Med 126.2 (January 15, 1997): 169-171.
PMID
9005754
Source
pubmed
Published In
Annals of internal medicine
Volume
126
Issue
2
Publish Date
1997
Start Page
169
End Page
171

Systemic lupus erythematosus. Diagnosis and treatment.

Systemic lupus erythematosus is a multisystem inflammatory disease characterized by antinuclear antibody production. The diagnosis of this disease is established on the basis of a constellation of clinical and serologic features. Therapy is directed to specific disease manifestations and involves an array of anti-inflammatory and immunosuppressive agents that are administered judiciously to limit toxicity.

Authors
Pisetsky, DS; Gilkeson, G; St Clair, EW
MLA Citation
Pisetsky, DS, Gilkeson, G, and St Clair, EW. "Systemic lupus erythematosus. Diagnosis and treatment." Med Clin North Am 81.1 (January 1997): 113-128. (Review)
PMID
9012757
Source
pubmed
Published In
Medical Clinics of North America
Volume
81
Issue
1
Publish Date
1997
Start Page
113
End Page
128

Specificity and immunochemical properties of antibodies to bacterial DNA.

DNA is a structurally heterogeneous molecule that elicits antibody production in both normal and aberrant immunity. In the prototypic autoimmune disease systemic lupus erythematosus, anti-DNA antibodies occur prominently and are serological markers for diagnosis and prognosis. These antibodies target conserved sites on both single-stranded (ss) and double-stranded (ds) DNA from essentially all species. In contrast, sera of normal human subjects (NHS) contain antibodies that selectively bind to DNA from certain bacteria. The NHS antibodies differ from lupus anti-DNA in their exclusive binding to foreign DNA as well as in their isotype, light chain distribution, and mode of DNA interaction. The properties of these antibodies suggest that they arise as a specific response to bacterial DNA introduced during infection or colonization. This conclusion is supported by studies demonstrating that bacterial DNA is a potent immunogen in normal mice and can induce antibodies specific for determinants on bacterial ss and dsDNA. Furthermore, immunization of normal animals with bacterial DNA elicits antibodies that bind mammalian as well as bacterial ssDNA; this immunization can also provoke glomerulonephritis. In addition to its immunogenicity, bacterial DNA is mitogenic, stimulating polyclonal activation of murine B cells; bacterial DNA can also induce cytokine production in the mouse. The range of immunological activities of bacterial DNA suggests an important role of this molecule in stimulating host defense in normal individuals and provoking autoimmunity in individuals predisposed to SLE.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Specificity and immunochemical properties of antibodies to bacterial DNA." Methods 11.1 (January 1997): 55-61.
PMID
8990089
Source
pubmed
Published In
Methods
Volume
11
Issue
1
Publish Date
1997
Start Page
55
End Page
61
DOI
10.1006/meth.1996.0387

CD19 regulates B lymphocyte signaling thresholds critical for the development of B-1 lineage cells and autoimmunity.

CD19 serves as a cell surface response regulator that establishes signaling thresholds critical for B lymphocyte development and activation. B lymphocytes from CD19-deficient mice are hyporesponsive to transmembrane signals, while B lymphocytes from mice that overexpress CD19 to even a small extent (25% increase) become hyperresponsive. The B-1 subpopulation of B lymphocytes is particularly sensitive to CD19 regulation, since their development is severely decreased in CD19-deficient mice. The effect of altered CD19 expression levels on the development of B cells was therefore examined using transgenic mice that express varying levels of cell surface CD19. In this study, immature B cells within the bone marrow of wild-type mice were found to specifically up-regulate CD19 expression levels by twofold as they mature, while CD5+ B cells within the spleen and peritoneum expressed even higher levels of CD19. The development of CD5+ B cells was severely decreased in CD19-deficient mice, while there was a linear correlation between increased CD19 expression levels and the increased frequency of CD5+ B cells within the peritoneum and spleen. In fact, CD5+ B cells became a major B lymphocyte population in mice that overexpressed CD19. Increased expression of CD19 also correlated with increased levels of endogenous anti-DNA Abs and rheumatoid factor. These results indicate that up-regulated expression of CD19 is functionally important for B cell development and that CD19 establishes signaling thresholds that regulate the generation of B-1 lymphocytes as well as the development of autoantibodies.

Authors
Sato, S; Ono, N; Steeber, DA; Pisetsky, DS; Tedder, TF
MLA Citation
Sato, S, Ono, N, Steeber, DA, Pisetsky, DS, and Tedder, TF. "CD19 regulates B lymphocyte signaling thresholds critical for the development of B-1 lineage cells and autoimmunity." J Immunol 157.10 (November 15, 1996): 4371-4378.
PMID
8906812
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
157
Issue
10
Publish Date
1996
Start Page
4371
End Page
4378

Interleukin-2 receptor levels in sera of patients with rheumatoid arthritis treated with sulfasalazine, parenteral gold, or placebo.

OBJECTIVE: To evaluate the associations of soluble serum interleukin-2 receptor (sIL-2R) levels in patients with rheumatoid arthritis (RA) with clinical and laboratory measures of disease activity and the predicted response to therapy. METHODS: sIL-2R levels were determined by ELISA in 137 patients with RA, not previously treated with 2nd line therapy. Patients were enrolled in a prospective, randomized, placebo controlled trial of sulfasalazine (SSZ) versus gold sodium thiomalate (GSTM), sponsored by the Cooperative Systematic Studies of Rheumatic Diseases. Using correlation analysis and regression modeling, the clinical utility of sIL-2R as a measure of disease activity and predictor of outcome was assessed. RESULTS: 91 women and 46 men with a mean age of 51 +/- 13 years and mean duration of disease of 64 +/- 78 months participated in this study. The mean sIL-2R level in all patients with RA was markedly elevated (980 +/- 589 U/ml) compared with that in healthy control subjects (446 +/- 196 U/ml; p = < 0.0001). There was no correlation between the sIL-2R levels and the joint pain/tenderness count, either at study entry or completion. There were significant positive correlations between the baseline sIL-2R and baseline erythrocyte sedimentation rate (ESR) and between the change in sIL-2R and the change in ESR. Both a multiple linear regression model and a multiple logistic regression model showed that baseline sIL-2R levels were not predictive of clinical outcome. CONCLUSION: sIL-2R levels are significantly elevated in patients with active RA and correlate positively with ESR. However, these results indicate that in patients with early RA, sIL-2R levels are neither associated with standard disease activity criteria nor predictive of the response to therapy with SSZ or GSTM, even after controlling for the simultaneous effects of other important clinical variables.

Authors
Merkel, PA; Dooley, MA; Dawson, DV; Pisetsky, DS; Polisson, RP
MLA Citation
Merkel, PA, Dooley, MA, Dawson, DV, Pisetsky, DS, and Polisson, RP. "Interleukin-2 receptor levels in sera of patients with rheumatoid arthritis treated with sulfasalazine, parenteral gold, or placebo." J Rheumatol 23.11 (November 1996): 1856-1861.
PMID
8923356
Source
pubmed
Published In
The Journal of rheumatology
Volume
23
Issue
11
Publish Date
1996
Start Page
1856
End Page
1861

Immune activation by bacterial DNA: a new genetic code.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Immune activation by bacterial DNA: a new genetic code." Immunity 5.4 (October 1996): 303-310. (Review)
PMID
8885863
Source
pubmed
Published In
Immunity
Volume
5
Issue
4
Publish Date
1996
Start Page
303
End Page
310

Activation of human B cells by phosphorothioate oligodeoxynucleotides.

To investigate the potential of DNA to elicit immune responses in man, we examined the capacity of a variety of oligodeoxynucleotides (ODNs) to stimulate highly purified T cell-depleted human peripheral blood B cells. Among 47 ODNs of various sequences tested, 12 phosphorothioate oligodeoxynucleotides (sODNs) induced marked B cell proliferation and Ig production. IL-2 augmented both proliferation and production of IgM, IgG, and IgA, as well as IgM anti-DNA antibodies, but was not necessary for B cell stimulation. Similarly, T cells enhanced stimulation, but were not necessary for B cell activation. After stimulation with the active sODNs, more than 95% of B cells expressed CD25 and CD86. In addition, B cells stimulated with sODNs expressed all six of the major immunoglobulin VH gene families. These results indicate that the human B cell response to sODN is polyclonal. Active sODN coupled to Sepharose beads stimulated B cells as effectively as the free sODN, suggesting that stimulation resulted from engagement of surface receptors. These data indicate that sODNs can directly induce polyclonal activation of human B cells in a T cell-independent manner by engaging as yet unknown B cell surface receptors.

Authors
Liang, H; Nishioka, Y; Reich, CF; Pisetsky, DS; Lipsky, PE
MLA Citation
Liang, H, Nishioka, Y, Reich, CF, Pisetsky, DS, and Lipsky, PE. "Activation of human B cells by phosphorothioate oligodeoxynucleotides." J Clin Invest 98.5 (September 1, 1996): 1119-1129.
PMID
8787674
Source
pubmed
Published In
Journal of Clinical Investigation
Volume
98
Issue
5
Publish Date
1996
Start Page
1119
End Page
1129
DOI
10.1172/JCI118894

Increased expression of blood mononuclear cell nitric oxide synthase type 2 in rheumatoid arthritis patients.

Nitric oxide (NO) is an important inflammatory mediator in nonhuman animal models of rheumatoid arthritis (RA). The purpose of the present study was to determine whether blood mononuclear cells from patients with active RA (as compared to control subjects) have higher levels of NO synthase type 2 (NOS2) and produce more NO in vitro. Leukocytes from 25 RA patients and 20 normal subjects were examined. Arthritis activity was assessed by tender and swollen joint counts, duration of morning stiffness, patient assessment of pain, physician and patient global assessment of disease activity, the modified Stanford Health Assessment Questionnaire, and by blood levels of acute phase reactants. Blood mononuclear cell NOS enzyme activity/antigen content and nitrite/nitrate formation in vitro were measured. Blood mononuclear cells from RA patients had increased NOS activity and increased NOS2 antigen content as compared to those from normal subjects, and responded to interferon-gamma with increased NOS expression and nitrite/nitrate production in vitro. NOS activity of freshly isolated blood mononuclear cells correlated significantly with disease activity, as assessed by render and swollen joint counts. Our results demonstrate that patients with RA have systemic activation for NOS2 expression, and that the degree of activation correlates with disease activity. Increased NOS2 expression and NO generation may be important in the pathogenesis of RA.

Authors
St Clair, EW; Wilkinson, WE; Lang, T; Sanders, L; Misukonis, MA; Gilkeson, GS; Pisetsky, DS; Granger, DI; Weinberg, JB
MLA Citation
St Clair, EW, Wilkinson, WE, Lang, T, Sanders, L, Misukonis, MA, Gilkeson, GS, Pisetsky, DS, Granger, DI, and Weinberg, JB. "Increased expression of blood mononuclear cell nitric oxide synthase type 2 in rheumatoid arthritis patients." J Exp Med 184.3 (September 1, 1996): 1173-1178.
PMID
9064335
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
184
Issue
3
Publish Date
1996
Start Page
1173
End Page
1178

Differences in V kappa gene utilization and VH CDR3 sequence among anti-DNA from C3H-lpr mice and lupus mice with nephritis.

To investigate the molecular properties of anti-DNA from lpr mice that express high levels of anti-DNA without immune-mediated nephritis, the sequences of VH and V kappa genes encoding 11 monoclonal anti-DNA antibodies derived from C3H-lpr/lpr (C3H-lpr) mice were studied. All of the C3H-lpr monoclonal anti-DNA bound single-stranded DNA while five also bound double-stranded DNA. Two of the hybridomas were clonally related as determined by Southern analysis and sequencing. Sequence analysis of C3H-lpr anti-DNA revealed the use of VH genes that encode anti-DNA from the MRL-lpr/lpr and (NZB X NZW) F1 mouse models of lupus, although differences occurred in the VH CDR3 amino acid content. In contrast, the V kappa genes from C3H-lpr mice lacked significant identity with previously reported V kappa genes for anti-DNA from lupus models. These results indicate that anti-DNA from C3H-lpr mice differ from anti-DNA from lupus mice with nephritis in patterns of V gene expression and suggest a molecular basis for the lack of pathogenicity of anti-DNA in these mice.

Authors
Wloch, MK; Alexander, AL; Pippen, AM; Pisetsky, DS; Gilkeson, GS
MLA Citation
Wloch, MK, Alexander, AL, Pippen, AM, Pisetsky, DS, and Gilkeson, GS. "Differences in V kappa gene utilization and VH CDR3 sequence among anti-DNA from C3H-lpr mice and lupus mice with nephritis." Eur J Immunol 26.9 (September 1996): 2225-2233.
PMID
8814271
Source
pubmed
Published In
European Journal of Immunology
Volume
26
Issue
9
Publish Date
1996
Start Page
2225
End Page
2233
DOI
10.1002/eji.1830260939

Final act.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Final act." Arthritis Care Res 9.4 (August 1996): 248-250.
PMID
8997908
Source
pubmed
Published In
Arthritis Care and Research
Volume
9
Issue
4
Publish Date
1996
Start Page
248
End Page
250

Modulation of renal disease in autoimmune NZB/NZW mice by immunization with bacterial DNA.

Preautoimmune New Zealand Black/White (NZB/NZW) mice immunized with Escherichia coli (EC) double standard (ds) DNA produce antibodies that bind mammalian dsDNA and display specificities similar to spontaneous lupus anti-DNA. Since calf thymus (CT) dsDNA fails to induce these antibodies, these results suggest a special potency of foreign DNA in inducing serological manifestations of lupus in a susceptible host. To assess the effects of DNA immunization on clinical manifestations in NZB/NZW mice, we measured renal disease and survival of mice immunized with either (a) EC dsDNA as complexes with methylated bovine serum albumin (mBSA) in adjuvant; (b) CT dsDNA with mBSA in adjuvant; (c)mBSA alone in adjuvant; or (d) unimmunized. After immunization with EC dsDNA, NZB/NZW mice developed significant levels of anti-dsDNA antibodies. Nevertheless, these mice had less proteinuria, nitrate/nitrite excretion, and glomerular pathology than mice immunized with either mBSA alone, CT dsDNA/mBSA complexes, or unimmunized mice. Survival of the EC dsDNA immunized mice was significantly increased compared with the other mice. Furthermore, immunization of mice after the onset of anti-DNA production and proteinuria stabilized nephritis and prolonged survival. The improvement in renal disease occurred despite the expression of autoantibodies that bound mammalian dsDNA as well as glomerular antigens. These results suggest that bacterial DNA has immunological properties that attenuate murine lupus despite the induction of pathogenic antibodies.

Authors
Gilkeson, GS; Ruiz, P; Pippen, AM; Alexander, AL; Lefkowith, JB; Pisetsky, DS
MLA Citation
Gilkeson, GS, Ruiz, P, Pippen, AM, Alexander, AL, Lefkowith, JB, and Pisetsky, DS. "Modulation of renal disease in autoimmune NZB/NZW mice by immunization with bacterial DNA." J Exp Med 183.4 (April 1, 1996): 1389-1397.
PMID
8666897
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
183
Issue
4
Publish Date
1996
Start Page
1389
End Page
1397

A multicenter trial of recombinant human interferon gamma in patients with systemic sclerosis: effects on cutaneous fibrosis and interleukin 2 receptor levels.

OBJECTIVE: To evaluate the acute toxicity, potential efficacy, and effects on the soluble interleukin 2 receptor (sIL-2R) of recombinant human interferon gamma (rIFN-gamma) in patients with systemic sclerosis (SSc). METHODS: A multicentered, pilot clinical trial of rIFN-gamma was performed on 20 patients (15 women, 5 men, mean age 45 years) with active cutaneous SSc (mean disease duration 36 months) to evaluate it potential as a novel therapy for this untreatable disorder. After one week of rIFN-gamma 0.01 mg/m2/day, subjects self-administered rIFN-gamma 0.1 mg/m2/day intramuscularly for a total of 18 weeks. The major outcome variable was a modified skin score (0 = normal skin, 3 = hidebound skin) measured and summed from 15 anatomic areas of the body. sIL-2R levels were measured by ELISA at entry and exit from the study. RESULTS: The clinical results were modest at best. Nine of 20 patients achieved at least a 20% reduction in skin score, with one patient showing almost total remission of all skin abnormalities. The mean skin score at entry for all subjects was 22.8 +/- 8.9 and over the course of the trial improved marginally compared to baseline (mean change -4.72 +/- 6.62; p = 0.008). However, 8 subjects did not change appreciably while in the trial. Antibodies to Scl-70 were observed in only 5 patients (all with diffuse scleroderma) and were not associated with either response to or complications from therapy. The adverse reactions were frequent and occasionally severe. Ten subjects were withdrawn because of exacerbation of Raynaud's symptoms (n = 5), constitutional symptoms (n = 2), development of renal crises (n = 2), and mild pancytopenia (n = 1). Minor laboratory abnormalities were common and included elevation of cholesterol, triglycerides, hepatic transaminases, and reduction in white blood cell count. Compared to controls, mean sIL-2R was markedly elevated at entry (1309 +/- 495 U/ml; p = 0.0001) and did not change appreciably at exit. Spearman correlation analysis showed a trend but no statistically significant association of skin score with sIL-2R (R = 0.408; p = 0.074). However, sIL-2R was significantly correlated with erythrocyte sedimentation rate (R = 0.542; p = 0.0165). A subset analysis revealed that skin score (p = 0.0001) and sIL-2R (p = 0.00170) were significantly higher at baseline for patients with diffuse scleroderma compared to patients with limited disease. CONCLUSION: rIFN-gamma may be beneficial for some patients with SSc, but the benefit appears marginal for most individuals and the side effects frequent. Although sIL-2R was elevated in many of the patients with SSc, it did not appear to be correlated with activity of cutaneous disease or response to therapy.

Authors
Polisson, RP; Gilkeson, GS; Pyun, EH; Pisetsky, DS; Smith, EA; Simon, LS
MLA Citation
Polisson, RP, Gilkeson, GS, Pyun, EH, Pisetsky, DS, Smith, EA, and Simon, LS. "A multicenter trial of recombinant human interferon gamma in patients with systemic sclerosis: effects on cutaneous fibrosis and interleukin 2 receptor levels." J Rheumatol 23.4 (April 1996): 654-658.
PMID
8730122
Source
pubmed
Published In
The Journal of rheumatology
Volume
23
Issue
4
Publish Date
1996
Start Page
654
End Page
658

A piece of my mind. Moments of love.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "A piece of my mind. Moments of love." JAMA 275.6 (February 14, 1996): 433-434.
PMID
8627953
Source
pubmed
Published In
JAMA : the journal of the American Medical Association
Volume
275
Issue
6
Publish Date
1996
Start Page
433
End Page
434

The breakthrough.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "The breakthrough." Ann Intern Med 124.3 (February 1, 1996): 345-347.
PMID
8554234
Source
pubmed
Published In
Annals of internal medicine
Volume
124
Issue
3
Publish Date
1996
Start Page
345
End Page
347

Restoring our health.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Restoring our health." Arthritis Care Res 9.1 (February 1996): 9-12.
PMID
8945107
Source
pubmed
Published In
Arthritis Care and Research
Volume
9
Issue
1
Publish Date
1996
Start Page
9
End Page
12

The immunologic properties of DNA.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "The immunologic properties of DNA." J Immunol 156.2 (January 15, 1996): 421-423. (Review)
PMID
8543788
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
156
Issue
2
Publish Date
1996
Start Page
421
End Page
423

Bacterial DNA induces murine interferon-gamma production by stimulation of interleukin-12 and tumor necrosis factor-alpha.

Bacterial, but not mammalian DNA, can induce interferon-gamma (IFN-gamma) in murine splenocytes. To elucidate the basis of this activity, we have assessed in vitro cytokine production by C3H/HeJ splenocytes stimulated with either DNA from Escherichia coli or a synthetic oligonucleotide containing an active palindromic sequence identified from DNA. Both DNAs induced IFN-gamma production, with the requirement for intact DNA shown by sensitivity to DNase digestion. Fractionated cell populations were evaluated to determine direct or indirect cellular effects of the DNA. Although bacterial DNA failed to induce IFN-gamma in the nonadherent cell population, supernatants from adherent cells stimulated by DNA induced IFN-gamma production by these cells. Interleukin-12 (IL-12) was detectable in supernatants from DNA-stimulated splenocytes before IFN-gamma, and neutralizing antibodies directed against IL-12 markedly inhibited the induction of IFN-gamma. Anti-tumor necrosis factor-alpha (TNF-alpha) antibodies also inhibited IFN-gamma production, and the combination of both anti-IL-12 and anti-TNF-alpha could totally inhibit production of IFN-gamma. Taken together, these results indicate that the stimulation of IFN-gamma production by bacterial DNA is mediated by IL-12 and TNF-alpha and point to macrophages/monocytes as targets of action of this macromolecule.

Authors
Halpern, MD; Kurlander, RJ; Pisetsky, DS
MLA Citation
Halpern, MD, Kurlander, RJ, and Pisetsky, DS. "Bacterial DNA induces murine interferon-gamma production by stimulation of interleukin-12 and tumor necrosis factor-alpha." Cell Immunol 167.1 (January 10, 1996): 72-78.
PMID
8548847
Source
pubmed
Published In
Cellular Immunology
Volume
167
Issue
1
Publish Date
1996
Start Page
72
End Page
78
DOI
10.1006/cimm.1996.0009

American Dream

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "American Dream." Journal of Medical Humanities 17.3 (1996): 203-211.
Source
scival
Published In
Journal of Medical Humanities
Volume
17
Issue
3
Publish Date
1996
Start Page
203
End Page
211

Human b cell activation induced by specific oligodeoxynucleotides

To investigate the potential of DNA to elicit immune responses, we examined the capacity of a variety of oligodeoxynucleotides (ODNs) to stimulate highly purified T cell depleted human peripheral blood B cells. Among 34 ODNs tested, three specific phosphorothioates (27bp antisense to rev gene of HIV, 21 bp antisense to HSV, and 20bp randomer) induced B cell proliferation and Ig production at low concentrations (1-5 ng/ml). IL2 augmented both proliferation and production of IgM, IgG and IgA, as well as IgM anti-DNA antibodies, but was not necessary for B cell stimulation. Similarly, intact T cells enhanced stimulation, but were not necessary for B cell activation. After stimulation with the active ODNs, more than 90% of B cells expressed CD25 and CD86. In addition, B cells stimulated with ODNs expressed all 6 of the major immune-globulin VH gene families. These results indicate that the human B cell response to ODNs is polyclonal. Active ODNs coupled to sepharose beads stimulated B cells as effectively as the free ODNs, indicating that stimulation resulted from engagement of surface receptors. These data indicate that specific ODNs can directly induce polyclonal activation of human B celts in a T cell-independent manner by engaging as yet unknown B cell surface receptors.

Authors
Liang, H; Nishioka, Y; Reich, C; Pisetsky, D; Lipsky, PE
MLA Citation
Liang, H, Nishioka, Y, Reich, C, Pisetsky, D, and Lipsky, PE. "Human b cell activation induced by specific oligodeoxynucleotides." FASEB Journal 10.6 (1996): A1207-.
Source
scival
Published In
The FASEB journal : official publication of the Federation of American Societies for Experimental Biology
Volume
10
Issue
6
Publish Date
1996
Start Page
A1207

Fact or fiction? [4] (multiple letters)

Authors
Twombly, R; Pisetsky, DS
MLA Citation
Twombly, R, and Pisetsky, DS. "Fact or fiction? [4] (multiple letters)." Annals of Internal Medicine 124.11 (1996): 1016-1017.
PMID
8624058
Source
scival
Published In
Annals of Internal Medicine
Volume
124
Issue
11
Publish Date
1996
Start Page
1016
End Page
1017

Consultation at twilight.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Consultation at twilight." N C Med J 56.12 (December 1995): 608-610.
PMID
8584058
Source
pubmed
Published In
North Carolina Medical Journal
Volume
56
Issue
12
Publish Date
1995
Start Page
608
End Page
610

Immunological properties of bacterial DNA.

Authors
Pisetsky, DS; Reich, C; Crowley, SD; Halpern, MD
MLA Citation
Pisetsky, DS, Reich, C, Crowley, SD, and Halpern, MD. "Immunological properties of bacterial DNA." Ann N Y Acad Sci 772 (November 27, 1995): 152-163. (Review)
PMID
8546388
Source
pubmed
Published In
Annals of the New York Academy of Sciences
Volume
772
Publish Date
1995
Start Page
152
End Page
163

The influence of variable-region somatic mutations on the specificity and pathogenicity of murine monoclonal anti-DNA antibodies.

Antibodies to DNA (anti-DNA) occur prominently in systemic lupus erythematosus and provoke inflammatory damage in the kidneys. To determine the factors that confer pathogenicity on antibodies of this specificity, we investigated the in vitro and in vivo glomerular binding by members of four clonally related sets of monoclonal anti-DNA antibodies from lupus mice. Somatic mutations within the clonal sets enhanced binding to double-stranded DNA (dsDNA). Binding to permeabilized glomeruli in vitro was observed among affinity-purified preparations of these antibodies independent of specificity for dsDNA. In normal mice injected with hybridoma cell lines, nephritis as assessed by histology and immunofluorescence did not correlate with antibody affinity for DNA. By multivariate analysis, in vitro glomerular binding was the most predictive parameter of histologic outcome. These findings indicate that somatic mutations occurring during maturation of the autoimmune response do not necessarily enhance pathogenicity.

Authors
Gilkeson, GS; Bernstein, K; Pippen, AM; Clarke, SH; Marion, T; Pisetsky, DS; Ruiz, P; Lefkowith, JB
MLA Citation
Gilkeson, GS, Bernstein, K, Pippen, AM, Clarke, SH, Marion, T, Pisetsky, DS, Ruiz, P, and Lefkowith, JB. "The influence of variable-region somatic mutations on the specificity and pathogenicity of murine monoclonal anti-DNA antibodies." Clin Immunol Immunopathol 76.1 Pt 1 (July 1995): 59-67.
PMID
7606869
Source
pubmed
Published In
Clinical Immunology and Immunopathology
Volume
76
Issue
1 Pt 1
Publish Date
1995
Start Page
59
End Page
67
DOI
10.1006/clin.1995.1088

The effects of short-term treatment with the prostaglandin E1 (PGE1) analog misoprostol on inflammatory mediator production in murine lupus nephritis.

MRL-lpr/lpr mice spontaneously develop an autoimmune disease with nephritis similar to human systemic lupus erythematosus. In these animals, treatment with E-series prostaglandins ameliorates renal disease and prolongs survival, perhaps by modulating production of cytokines or eicosanoids. To further define the mechanisms of action of E-series prostaglandins in established murine lupus nephritis, we administered the prostaglandin E1 (PGE1) analog misoprostol to 20-week-old MRL-lpr/lpr mice by twice-daily subcutaneous injection. After 2 days of treatment, misoprostol reduced renal cortical interleukin-1 beta (IL-1 beta) mRNA levels compared to vehicle-treated controls (0.19 +/- 0.06 (misoprostol) vs 0.50 +/- 0.04 (vehicle) densitometry units; P < 0.005). A similar reduction in cortical IL-1 beta mRNA levels was found in left kidneys harvested from MRL-lpr/lpr mice following 2 days of treatment with misoprostol compared to right kidneys harvested from the same animal prior to the first dose of PGE1 analog (0.12 +/- .05 (left) vs 0.39 +/- 0.18 (right) densitometry units; P < 0.05). This reduction in cortical IL-1 beta mRNA levels was not associated with alterations in renal production of thromboxane B2, PGE2, or leukotriene B4 or with significant changes in the severity of renal inflammatory cell infiltrates. Time-course studies indicated that IL-1 beta mRNA levels were decreased within 24 hr of initiating misoprostol therapy. This reduction in IL-1 beta mRNA levels was transient because levels were not reduced after 1 week of treatment with the PGE1 analog.(ABSTRACT TRUNCATED AT 250 WORDS)

Authors
Fan, PY; Ruiz, P; Pisetsky, DS; Spurney, RF
MLA Citation
Fan, PY, Ruiz, P, Pisetsky, DS, and Spurney, RF. "The effects of short-term treatment with the prostaglandin E1 (PGE1) analog misoprostol on inflammatory mediator production in murine lupus nephritis." Clin Immunol Immunopathol 75.2 (May 1995): 125-130.
PMID
7704969
Source
pubmed
Published In
Clinical Immunology and Immunopathology
Volume
75
Issue
2
Publish Date
1995
Start Page
125
End Page
130

Induction of cross-reactive anti-dsDNA antibodies in preautoimmune NZB/NZW mice by immunization with bacterial DNA.

To investigate the role of antigen drive in anti-double-stranded (ds) DNA production, the antibody response induced in lupus-prone NZB/NZW mice by E. coli (EC) dsDNA was evaluated. Preautoimmune NZB/NZW female mice were immunized with complexes of EC dsDNA with methylated bovine serum albumin (mBSA) in complete Freund's adjuvant; control mice received either mBSA complexes with calf thymus (CT) dsDNA or mBSA alone in adjuvant. IgG antibody responses were assessed by ELISA. Similar to normal mice, immunized NZB/NZW mice produced significant levels of anti-dsDNA when measured with EC dsDNA as antigen. Whereas normal mice produce antibodies which are specific for the immunizing bacterial DNA, NZB/NZW mice produced antibodies that bound crossreactively to CT dsDNA by ELISA. Furthermore, the induced antibodies resembled lupus anti-DNA in their fine specificity for polynucleotide antigens and reactivity with Crithidia luciliae DNA. Despite their response to EC dsDNA, NZB/NZW mice immunized with CT dsDNA failed to generate significant anti-dsDNA responses. These results provide further evidence for the enhanced immunogenicity of bacterial DNA and suggest that immune cell abnormalities in NZB/NZW mice promote the generation of crossreactive autoantibody responses when confronted with a foreign DNA.

Authors
Gilkeson, GS; Pippen, AM; Pisetsky, DS
MLA Citation
Gilkeson, GS, Pippen, AM, and Pisetsky, DS. "Induction of cross-reactive anti-dsDNA antibodies in preautoimmune NZB/NZW mice by immunization with bacterial DNA." J Clin Invest 95.3 (March 1995): 1398-1402.
PMID
7883986
Source
pubmed
Published In
Journal of Clinical Investigation
Volume
95
Issue
3
Publish Date
1995
Start Page
1398
End Page
1402
DOI
10.1172/JCI117793

In vitro inhibition of murine IFN gamma production by phosphorothioate deoxyguanosine oligomers.

Phosphorothioate (PT) oligonucleotides are designed as specific agents for antisense therapy although they have been reported to exert non-specific immunomodulatory effects. To elucidate further their actions, the effect of PT deoxyguanosine oligomers (S-oligo(dG)) on in vitro cytokine production by mouse splenocytes was studied. S-oligo(dG)20 inhibited production of INF gamma induced by Con A, E. coli DNA or the combination of PMA and calcium ionophore A23187. The diester analogue was inactive, and of PT homo-oligomers tested, S-oligo(dG)20 was the most active. PT compounds with as few as 5 dG residues could also block INF gamma production. These results indicate that base composition and length, as well as the PT backbone, contribute to the inhibition of INF gamma production and extend the range of immunomodulatory effects of PT compounds.

Authors
Halpern, MD; Pisetsky, DS
MLA Citation
Halpern, MD, and Pisetsky, DS. "In vitro inhibition of murine IFN gamma production by phosphorothioate deoxyguanosine oligomers." Immunopharmacology 29.1 (February 1995): 47-52.
PMID
7768671
Source
pubmed
Published In
Immunopharmacology
Volume
29
Issue
1
Publish Date
1995
Start Page
47
End Page
52

Immunologic consequences of nucleic acid therapy.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Immunologic consequences of nucleic acid therapy." Antisense Res Dev 5.3 (1995): 219-225. (Review)
PMID
8785478
Source
pubmed
Published In
Antisense Research and Development
Volume
5
Issue
3
Publish Date
1995
Start Page
219
End Page
225

Letters to the editor [1]

Authors
Barrera, P; Ward, MM; Pyun, E; Pisetsky, DS
MLA Citation
Barrera, P, Ward, MM, Pyun, E, and Pisetsky, DS. "Letters to the editor [1]." Clinical Immunology and Immunopathology 76.1 I (1995): 102-104.
Source
scival
Published In
Clinical Immunology and Immunopathology
Volume
76
Issue
1 I
Publish Date
1995
Start Page
102
End Page
104
DOI
10.1006/clin.1995.1094

The mechanism of autoantibody production in an autoimmune MRL/lpr mouse.

Rheumatoid factors (RF) and anti-DNA Abs from MRL/lpr mice have features similar to Abs directed toward foreign Ags, indicating a role of specific activation by Ags during disease. But our previous studies and analogous studies from others concentrated on a limited subset of hybridomas selected on the basis of Ag binding to well characterized target autoantigens. Thus, it has been unclear to what extent clonal expansion is restricted to identifiable autospecificities. To obtain a more complete picture of disease-associated autoantibody production, we designed the following experiment. A large number of B cell hybridomas were generated from the spleen of an MRL/lpr mouse and then analyzed for self-specificity, sequence, and clonal relationship. Surprisingly, we found that clonal expansion was limited to only a few autospecificities, implying a unique property of this response. In addition, we used Southern blotting with heavy and L chain constant region probes to screen both RF and non-RF hybridomas for membership in clones, one of which was first identified among RF hybridomas. We found no non-RF members of this clone. The size and number of mutations of this clone were sufficient for us to conclude that nonspecific (i.e., non-RF) mutant members are rapidly lost. Had an Ag other than IgG2a been driving clonal expansion, we should have seen mutants that retained spectificity for that Ag but that lost specificity for IgG2a. This observation, along with the restriction of clonal expansion to a few autospecificities, provides strong evidence that normal autoantigens themselves drive autoantibody clonal expansion.

Authors
Shan, H; Shlomchik, MJ; Marshak-Rothstein, A; Pisetsky, DS; Litwin, S; Weigert, MG
MLA Citation
Shan, H, Shlomchik, MJ, Marshak-Rothstein, A, Pisetsky, DS, Litwin, S, and Weigert, MG. "The mechanism of autoantibody production in an autoimmune MRL/lpr mouse." J Immunol 153.11 (December 1, 1994): 5104-5120.
PMID
7525723
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
153
Issue
11
Publish Date
1994
Start Page
5104
End Page
5120

Serial measurement of serum interleukin-2 receptor levels in patients with rheumatoid arthritis: limited evidence for a role of T cell activation in clinical exacerbations.

To investigate the association of T cell activation with clinical exacerbations of RA, we measured serum levels of soluble interleukin-2 receptors (sIL2R), a marker of T cell activation, in serial samples obtained from 23 patients with RA. sIL2R measurements were performed on sera obtained from each patient every 2 weeks for up to 60 weeks, and levels were correlated with swollen joint counts, tender joint counts, physician global assessments, patient global assessments, pain scores, Health Assessment Questionnaire Disability Index scores, and Westergren erythrocyte sedimentation rates measured simultaneously. There were no significant correlations between changes in sIL2R levels and changes in any of the other measures, nor were lead-lag relationships detected, for the group as a whole. Examination of the time courses of individual patients revealed significant positive correlations between changes in sIL2R levels and changes in swollen joint counts in five patients; significant correlations with other measures were present in three or fewer patients. sIL2R levels also varied little over the 2-week time interval of greatest clinical change in each patient. These results suggest either that clinical exacerbations of RA are not associated with changes in T cell activation or that sIL2R levels do not accurately reflect such changes.

Authors
Ward, MM; Pyun, E; Pisetsky, DS
MLA Citation
Ward, MM, Pyun, E, and Pisetsky, DS. "Serial measurement of serum interleukin-2 receptor levels in patients with rheumatoid arthritis: limited evidence for a role of T cell activation in clinical exacerbations." Clin Immunol Immunopathol 73.3 (December 1994): 296-304.
PMID
7661913
Source
pubmed
Published In
Clinical Immunology and Immunopathology
Volume
73
Issue
3
Publish Date
1994
Start Page
296
End Page
304

Fish oil feeding modulates leukotriene production in murine lupus nephritis.

Diets enriched with fish oil (FO) ameliorate kidney disease in the MRL-lpr/lpr murine model of lupus nephritis. Although the mechanisms of this effect are not known, FO is rich in the polyunsaturated fatty acid eicosapentaenoic acid (EPA) which may have profound effects on eicosanoid metabolism. In MRL-lpr/lpr mice, FO feeding reduces renal production of cyclooxygenase metabolites. However, EPA may also affect the metabolism of arachidonate by the 5-lipoxygenase (5-LO) pathway and enhanced production of 5-LO metabolites has been implicated in the pathogenesis of kidney disease in MRL-lpr/lpr mice. We therefore investigated the effects of FO feeding on production of 5-LO metabolites in 20 week old MRL-lpr/lpr mice. After 8 weeks of dietary supplementation with FO, both renal hemodynamic function and glomerular histology were improved compared to safflower oil (SO) controls. Amelioration of kidney disease was associated with alterations in the pattern of leukotriene production by macrophages and kidneys from FO fed mice. There was a significant decrease in the production of leukotriene B4 (LTB4) and tetraene peptidoleukotrienes by peritoneal macrophages isolated from mice given FO compared to control animals. Similarly, dietary supplementation with FO decreased renal production of LTB4. Reduced production of tetraene leukotrienes was accompanied by a modest increase in the production of pentaene leukotrienes by macrophages from FO fed mice. We speculate that this modulation of leukotriene production by FO feeding may have beneficial effects on renal disease in autoimmune nephritis.

Authors
Spurney, RF; Ruiz, P; Albrightson, CR; Pisetsky, DS; Coffman, TM
MLA Citation
Spurney, RF, Ruiz, P, Albrightson, CR, Pisetsky, DS, and Coffman, TM. "Fish oil feeding modulates leukotriene production in murine lupus nephritis." Prostaglandins 48.5 (November 1994): 331-348.
PMID
7855311
Source
pubmed
Published In
Prostaglandins
Volume
48
Issue
5
Publish Date
1994
Start Page
331
End Page
348

Characterization of antibodies to bacterial double-stranded DNA in the sera of normal human subjects.

To assess the human antibody response to bacterial double-stranded (ds) DNA, sera from normal human subjects (NHS) were tested by ELISA for binding to highly purified dsDNA from Micrococcus lysodeikticus (MC). Of 38 NHS tested, 19 demonstrated significant activity to MC dsDNA (defined as an OD380 > 1.0 at a 1:100 dilution of serum) without appreciable binding to dsDNA from Escherichia coli or calf thymus. In competition ELISA, antibodies in NHS to MC dsDNA showed equivalent inhibition by MC ssDNA or dsDNA. Antibody specificity was further evaluated by testing the effects of ionic strength on binding. By ELISA, antibodies in NHS had greater binding to MC dsDNA at high ionic strengths than those in systemic lupus erythematosus sera. These results indicate that NHS have antibodies which bind bacterial dsDNA and extend the range of DNA determinants that can be recognized as foreign by the normal immune system.

Authors
Bunyard, MP; Pisetsky, DS
MLA Citation
Bunyard, MP, and Pisetsky, DS. "Characterization of antibodies to bacterial double-stranded DNA in the sera of normal human subjects." Int Arch Allergy Immunol 105.2 (October 1994): 122-127.
PMID
7920012
Source
pubmed
Published In
International archives of allergy and immunology
Volume
105
Issue
2
Publish Date
1994
Start Page
122
End Page
127

The influence of DNA size on the binding of anti-DNA antibodies in the solid and fluid phase.

To elucidate the interaction of anti-DNA antibodies with DNA, the reactivity of lupus sera with single-stranded fragments from calf thymus, Escherichia coli, and salmon testes DNA was investigated. These fragments were generated by digestion with the restriction enzyme HinfI and ranged in size from approximately 100-4000 bases. By ELISA using polystyrene microtiter plates, fragments from all three species were weakly antigenic compared to intact DNA. These fragments, however, were all antigenic when tested as inhibitors in competition-binding assays. The weak antigenicity of fragments could not be explained by poor adherence to the plates since fragments and intact DNA showed similar levels of binding as assessed using biotinylated preparations. Together, these results demonstrate that the antigenicity of DNA fragments is dramatically altered by solid-phase binding and suggest that constraints on topological or conformational rearrangements of DNA in the solid phase limit antibody interaction.

Authors
Pisetsky, DS; Reich, CF
MLA Citation
Pisetsky, DS, and Reich, CF. "The influence of DNA size on the binding of anti-DNA antibodies in the solid and fluid phase." Clin Immunol Immunopathol 72.3 (September 1994): 350-356.
PMID
7520375
Source
pubmed
Published In
Clinical Immunology and Immunopathology
Volume
72
Issue
3
Publish Date
1994
Start Page
350
End Page
356

The anti-La response of a single MRL/Mp-lpr/lpr mouse: specificity for DNA and VH gene usage.

Autoantibodies to ribonucleoproteins (RNP) occur prominently in human systemic lupus erythematosus and murine lupus models. In previous studies we demonstrated a relationship in MRL/Mp-lpr/lpr (MRL/lpr) mice between antibodies to Sm, an RNP autoantigen, and antibodies to DNA. Thus, many anti-Sm monoclonals bound DNA and expressed the same V region genes as anti-DNA. In addition, many had multiple VHCDR3 Arg residues suggestive of selection by DNA, and some had somatic mutations suggesting selection for mutant B cells by DNA. To determine whether autoantibodies to other RNP antigens are also associated with the anti-DNA response, we have analyzed the response to the La RNP. Six anti-La B cell hybridomas were generated from a single MRL/lpr mouse. Southern blot analysis of Ig V gene rearrangements and V gene sequences indicated two clonally related pairs, suggesting an oligoclonal response. Antibodies from all six hybridomas bound single-stranded DNA, while antibodies from five hybridomas bound double-stranded DNA. Two hybridomas expressed a VH7183 gene which is used by members of two previously reported anti-DNA clones and two anti-Sm/DNA clones of MRL/lpr origin. These data demonstrate an association between the anti-La and anti-DNA responses in MRL/lpr mice, suggesting that cross-reactive anti-RNP and anti-DNA responses are a general feature of autoimmunity in this lupus model.

Authors
Bloom, DD; St Clair, EW; Pisetsky, DS; Clarke, SH
MLA Citation
Bloom, DD, St Clair, EW, Pisetsky, DS, and Clarke, SH. "The anti-La response of a single MRL/Mp-lpr/lpr mouse: specificity for DNA and VH gene usage." Eur J Immunol 24.6 (June 1994): 1332-1338.
PMID
8206093
Source
pubmed
Published In
European Journal of Immunology
Volume
24
Issue
6
Publish Date
1994
Start Page
1332
End Page
1338
DOI
10.1002/eji.1830240614

The role of nitric oxide in the pathogenesis of spontaneous murine autoimmune disease: increased nitric oxide production and nitric oxide synthase expression in MRL-lpr/lpr mice, and reduction of spontaneous glomerulonephritis and arthritis by orally administered NG-monomethyl-L-arginine.

MRL-lpr/lpr mice spontaneously develop various manifestations of autoimmunity including an inflammatory arthropathy and immune complex glomerulonephritis. This study examines the role of nitric oxide, a molecule with proinflammatory actions, in the pathogenesis of MRL-lpr/lpr autoimmune disease. MRL-lpr/lpr mice excreted more urinary nitrite/nitrate (an in vivo marker of nitric oxide production) than did mice of normal strains and MRL-(+/+) and B6-lpr/lpr congenic strains. In addition, MRL-lpr/lpr peritoneal macrophages had an enhanced capacity to produce nitric oxide in vitro as well as increased nitric oxide synthase activity, and certain tissues from MRL-lpr/lpr mice had increased expression of inducible nitric oxide synthase (NOS) mRNA and increased amounts of material immunoreactive for inducible NOS. Oral administration of NG-monomethyl-L-arginine, a nitric oxide synthase inhibitor, prevented the development of glomerulonephritis and reduced the intensity of inflammatory arthritis in MRL-lpr/lpr mice. By using interspecific backcross mice, the gene for inducible NOS (Nosi) was mapped to mouse chromosome 11. This chromosomal localization was different from those loci that we have previously demonstrated to be linked to enhanced susceptibility to renal disease in an MRL-lpr/lpr cross. However, the chromosomal location of the NOS gene was consistent with an insulin-dependent diabetes locus identified in an analysis of nonobese diabetic (NOD) mice. These results suggest that elevated nitric oxide production could be important in the pathogenesis of autoimmunity, and that treatments to block the production of nitric oxide or block its effects might be valuable therapeutically.

Authors
Weinberg, JB; Granger, DL; Pisetsky, DS; Seldin, MF; Misukonis, MA; Mason, SN; Pippen, AM; Ruiz, P; Wood, ER; Gilkeson, GS
MLA Citation
Weinberg, JB, Granger, DL, Pisetsky, DS, Seldin, MF, Misukonis, MA, Mason, SN, Pippen, AM, Ruiz, P, Wood, ER, and Gilkeson, GS. "The role of nitric oxide in the pathogenesis of spontaneous murine autoimmune disease: increased nitric oxide production and nitric oxide synthase expression in MRL-lpr/lpr mice, and reduction of spontaneous glomerulonephritis and arthritis by orally administered NG-monomethyl-L-arginine." J Exp Med 179.2 (February 1, 1994): 651-660.
PMID
7507509
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
179
Issue
2
Publish Date
1994
Start Page
651
End Page
660

Interleukin-2 receptor levels in the sera of rheumatoid arthritis patients treated with methotrexate.

OBJECTIVE: To evaluate the association of the level of soluble serum interleukin-2 receptor (sIL-2R) with disease activity and response to therapy in patients with rheumatoid arthritis (RA). METHODS: The sIL-2R levels of 148 patients with refractory RA were determined by enzyme-linked immunosorbent assay. This parameter was correlated with other clinical observations obtained during a prospective, randomized, placebo-controlled trial of methotrexate, sponsored by the Cooperative Systematic Studies of Rheumatic Diseases consortium. Using statistical modeling, the usefulness of sIL-2R as a measure of disease activity and a predictor of outcome was evaluated. RESULTS: The mean sIL-2R level in all RA patients was markedly elevated compared with that in normal control subjects, and decreased significantly during the trial. There was no correlation of the sIL-2R level and the joint pain/tenderness count either at study entry or study end. There was a significant correlation of the sIL-2R level and the erythrocyte sedimentation rate, both at study entry and study end. A multiple linear regression model showed that treatment with methotrexate, but not the sIL-2R level or the change in sIL-2R level, predicted a change in joint count. A stepwise multiple logistic regression model defined no significant predictive information for outcome for the level of sIL-2R at study entry. CONCLUSION: After controlling for the simultaneous effects of other important clinical variables, the level of sIL-2R does not appear to predict the response to methotrexate in patients with refractory RA. Further analysis of cohorts of patients with earlier RA needs to be performed.

Authors
Polisson, RP; Dooley, MA; Dawson, DV; Pisetsky, DS
MLA Citation
Polisson, RP, Dooley, MA, Dawson, DV, and Pisetsky, DS. "Interleukin-2 receptor levels in the sera of rheumatoid arthritis patients treated with methotrexate." Arthritis Rheum 37.1 (January 1994): 50-56.
PMID
8129764
Source
pubmed
Published In
Arthritis and Rheumatism
Volume
37
Issue
1
Publish Date
1994
Start Page
50
End Page
56

Stimulation of murine lymphocyte proliferation by a phosphorothioate oligonucleotide with antisense activity for herpes simplex virus.

To investigate further the immunological properties of nucleic acids, the mitogenicity of a phosphorothioate oligonucleotide (S-oligo 1082) with antisense activity for herpes simplex virus was tested. This compound stimulated proliferation and antibody production by murine lymphocytes in in vitro cultures. Proliferation was dose-dependent and unaffected by T cell depletion. Furthermore, inclusion of a non-mitogenic DNA in the medium did not block stimulation. Since 1082 does not have homology to a known gene involved in lymphocyte activation, these observations suggest that S-oligo antisense compounds may display non-specific activating effects, at least on murine B cells.

Authors
Pisetsky, DS; Reich, CF
MLA Citation
Pisetsky, DS, and Reich, CF. "Stimulation of murine lymphocyte proliferation by a phosphorothioate oligonucleotide with antisense activity for herpes simplex virus." Life Sci 54.2 (1994): 101-107.
PMID
8277816
Source
pubmed
Published In
Life Sciences
Volume
54
Issue
2
Publish Date
1994
Start Page
101
End Page
107

Antinuclear antibodies

ANAs are important serologic markers of the rheumatic diseases, although their role in pathogenesis has been difficult to conceptualize because their target antigens are ubiquitous among cells and seemingly well protected in the cell interior. Future directions of research will include the identification of ANA specificities most highly correlated with disease manifestations and the development of immunoassays that provide more accurate information for assessing diagnosis, prognosis, and pathogenicity.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Antinuclear antibodies." Immunology and Allergy Clinics of North America 14.2 (1994): 371-385.
Source
scival
Published In
Immunology and Allergy Clinics of North America
Volume
14
Issue
2
Publish Date
1994
Start Page
371
End Page
385

DNA vaccination. A clue to memory?

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "DNA vaccination. A clue to memory?." Hum Immunol 38.4 (December 1993): 241-242.
PMID
8138418
Source
pubmed
Published In
Human Immunology
Volume
38
Issue
4
Publish Date
1993
Start Page
241
End Page
242

The effects of nonsteroidal antiinflammatory drug therapy in early rheumatoid arthritis on serum levels of soluble interleukin 2 receptor, CD4, and CD8.

OBJECTIVE: Cell surface molecules can be shed by activated T lymphocytes and measured in serum to assess in vivo T cell activation. To evaluate the relationship between these serum markers and disease activity in rheumatoid arthritis (RA), we determined levels of soluble interleukin 2 receptor (sIL-2R), CD4 (sCD4), and CD8 (sCD8) in sera from a well characterized group of 26 patients with active RA treated with a nonsteroidal antiinflammatory drug (NSAID) alone. METHODS: A retrospective, blinded determination of sIL-2R, sCD4, and sCD8 levels in serum samples from patients with early, active RA participating in 2 trials of NSAID therapy. Commercially available enzyme linked immunosorbent assays were employed. Data analysis included nonparametric techniques and correction for multiple comparisons. RESULTS: The patients with RA had significantly elevated levels of sIL-2R at baseline compared with age matched healthy controls. During NSAID therapy, mean sIL-2R levels among responders decreased to lower levels while nonresponder levels increased, although these trends did not reach statistical significance. Patients with RA did not differ from controls in baseline measures of serum sCD4 or sCD8 levels. Moreover, the serum sIL-2R, sCD4, and sCD8 levels among patients did not vary significantly from their baseline measures during NSAID therapy, irrespective of response. CONCLUSION: Our results suggest that elevated levels of serum sIL-2R in early RA likely reflect generalized immune system activation, not always associated with elevated serum sCD4 or sCD8 levels or varying with other measures of disease activity in RA.

Authors
Dooley, MA; Cush, JJ; Lipsky, PE; Dawson, DV; Pisetsky, DS
MLA Citation
Dooley, MA, Cush, JJ, Lipsky, PE, Dawson, DV, and Pisetsky, DS. "The effects of nonsteroidal antiinflammatory drug therapy in early rheumatoid arthritis on serum levels of soluble interleukin 2 receptor, CD4, and CD8." J Rheumatol 20.11 (November 1993): 1857-1862.
PMID
8308770
Source
pubmed
Published In
The Journal of rheumatology
Volume
20
Issue
11
Publish Date
1993
Start Page
1857
End Page
1862

Stimulation of in vitro proliferation of murine lymphocytes by synthetic oligodeoxynucleotides.

To elucidate the properties of mitogenic nucleic acids, the ability of oligodeoxynucleotides to stimulate the in vitro proliferation of murine lymphocytes was investigated. The compounds tested were a series of oligodeoxynucleotides, synthesized with either phosphodiester or phosphorothioate chemistry and containing (dG) and (dC) alone or together. Among oligodeoxynucleotides tested, phosphorothioates were more active than phosphodiesters and stimulated thymidine incorporation under the same conditions as mitogenic non-mammalian DNA. Mitogenesis was unaffected by depletion of T cells, suggesting B cells as the predominant cell type stimulated. These results indicate that mitogenic nucleic acids need not have an extended polymeric structure and raise the possibility that antisense compounds have immunologic activity, at least in animal models.

Authors
Pisetsky, DS; Reich, C
MLA Citation
Pisetsky, DS, and Reich, C. "Stimulation of in vitro proliferation of murine lymphocytes by synthetic oligodeoxynucleotides." Mol Biol Rep 18.3 (October 1993): 217-221.
PMID
8114689
Source
pubmed
Published In
Molecular Biology Reports
Volume
18
Issue
3
Publish Date
1993
Start Page
217
End Page
221

Induction of immune-mediated glomerulonephritis in normal mice immunized with bacterial DNA.

Normal mice immunized with bacterial DNA produce high titers of anti-DNA antibodies and represent a new model for autoantibody production in systemic lupus erythematosus. To determine whether DNA immunization can also provoke clinical manifestations of lupus, the occurrence of nephritis in immunized mice was assessed and correlated with levels of anti-DNA as well as antibodies to glomerular antigens. BALB/c mice immunized with Escherichia coli single-stranded DNA in complexes with methylated bovine serum albumin in adjuvant showed increased proteinuria compared to control mice immunized with mBSA alone. Furthermore, DNA immunized mice had significantly greater glomerular proliferative changes and immunoglobulin deposition than control mice. In an in vitro assay, sera from DNA immunized mice exhibited greater binding to glomerular antigens than sera from control mice. Compared to sera, renal eluates from DNA-immunized mice were enriched for anti-DNA and glomerular binding activity. These data indicate that immunization of normal mice with E. coli DNA induces an immune-mediated proliferative glomerulonephritis that is likely secondary to the renal deposition of anti-DNA antibodies.

Authors
Gilkeson, GS; Ruiz, P; Howell, D; Lefkowith, JB; Pisetsky, DS
MLA Citation
Gilkeson, GS, Ruiz, P, Howell, D, Lefkowith, JB, and Pisetsky, DS. "Induction of immune-mediated glomerulonephritis in normal mice immunized with bacterial DNA." Clin Immunol Immunopathol 68.3 (September 1993): 283-292.
PMID
8370182
Source
pubmed
Published In
Clinical Immunology and Immunopathology
Volume
68
Issue
3
Publish Date
1993
Start Page
283
End Page
292

Autoantibodies and their significance.

In systemic lupus erythematosus, autoantibodies have structural features that indicate in vivo selection by a T cell-dependent, antigen-driven process. The B-cell component of these responses resembles a conventional antibody response, whereas the T-cell component may involve diverse stimulatory mechanisms and levels of regulatory control. Characterizing T-cell epitopes of autoantigens has been difficult because these molecules are ubiquitous and exist inside the cells as multicomponent, macromolecular complexes. Autoantibodies can mediate disease manifestations by various mechanisms, with variable region structures determining the pattern and severity of tissue injury.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Autoantibodies and their significance." Curr Opin Rheumatol 5.5 (September 1993): 549-556. (Review)
PMID
8398606
Source
pubmed
Published In
Current Opinion in Rheumatology
Volume
5
Issue
5
Publish Date
1993
Start Page
549
End Page
556

Molecular characterization of anti-DNA antibodies induced in normal mice by immunization with bacterial DNA. Differences from spontaneous anti-DNA in the content and location of VH CDR3 arginines.

Immunization of normal mice with bacterial DNA induces a significant anti-DNA response that includes antibodies resembling some lupus anti-DNA in their binding properties, although lacking specificity for mammalian dsDNA. To determine the structure of these induced antibodies and their relationship to anti-DNA from lupus mice, we have characterized the clonality and selected V-region sequences of a panel of 20 anti-DNA antibodies from 3 BALB/c mice immunized with ssDNA from Escherichia coli. Southern blot analysis of H and L chain rearrangements indicated that two of the animals expressed pairs of clonally related antibodies. Amino acid sequences of 10 of the induced antibodies demonstrated predominant utilization of J558 family VH genes and JH4 in association with various DH, J kappa and V kappa genes. Among the VH CDR3 of these 10 antibodies, 4 displayed arginine residues as a result of N region additions. None of these antibodies, however, had more than one arginine residue in VH CDR3 nor arginines at positions 100 or 100a, characteristic features of lupus antibodies to dsDNA. These results suggest that normal mice immunized with bacterial DNA display certain facets of DNA Ag drive, although lacking the mechanisms for the production of antibodies to mammalian dsDNA.

Authors
Gilkeson, GS; Bloom, DD; Pisetsky, DS; Clarke, SH
MLA Citation
Gilkeson, GS, Bloom, DD, Pisetsky, DS, and Clarke, SH. "Molecular characterization of anti-DNA antibodies induced in normal mice by immunization with bacterial DNA. Differences from spontaneous anti-DNA in the content and location of VH CDR3 arginines." J Immunol 151.3 (August 1, 1993): 1353-1364.
PMID
8335932
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
151
Issue
3
Publish Date
1993
Start Page
1353
End Page
1364

Effect of sex on the induction of anti-DNA antibodies in normal mice immunized with bacterial DNA.

Immunization of normal mice with bacterial DNA elicits a significant IgG anti-DNA response and has been explored as a model of systemic lupus erythematosus. To determine whether this induced response is influenced by sex, we have measured anti-DNA levels in normal male and female BALB/c mice immunized with single stranded DNA from E. coli as complexes with methylated bovine serum albumin (mBSA) in adjuvant. By ELISA assays, anti-DNA levels of immunized females were approximately 16-fold higher than those of immunized males; levels of antibodies to the mBSA carrier were similar, however. The antibodies from females and males showed a similar degree of cross-reactivity when assayed using other natural and synthetic DNA antigens, including mammalian DNA. These findings suggest the potentiation of anti-DNA production in females by antigen-specific mechanisms and provide further evidence that immunization with bacterial DNA replicates features of autoantibody production in SLE.

Authors
Palmer, SM; Gilkeson, GS; Pisetsky, DS
MLA Citation
Palmer, SM, Gilkeson, GS, and Pisetsky, DS. "Effect of sex on the induction of anti-DNA antibodies in normal mice immunized with bacterial DNA." Lupus 2.4 (August 1993): 251-255.
PMID
8268973
Source
pubmed
Published In
Lupus
Volume
2
Issue
4
Publish Date
1993
Start Page
251
End Page
255
DOI
10.1177/096120339300200408

Murine monoclonal antibodies specific for conserved and non-conserved antigenic determinants of the human and murine Ku autoantigens.

The Ku autoantigen is a DNA binding factor consisting of 70 and approximately 80 kDa proteins (p70 and p80, respectively) which form a heterodimer. The p70/p80 dimer appears to be crucial for the function of a 350 kDa DNA-dependent protein kinase (DNA-PK) that phosphorylates certain transcription factors in vitro. Previous studies have suggested that Ku is abundant in primate cells, but undetectable in most non-primate cells. However, it is unclear if this reflects low abundance of Ku (and possibly DNA-PK activity) in non-primate cells, a lack of antibodies crossreactive with non-primate Ku proteins, or both. Ku was first identified with human autoimmune sera, but the suitability of these sera for studying the distribution, abundance and function of Ku is limited by the polyclonal immune response to Ku and the presence of contaminating autoantibodies in most patients' sera. In the present studies, we determined the specificities of murine anti-Ku monoclonal antibodies (mAbs) using cellular Ku as well as recombinant human and murine Ku antigens. Immunofluorescence studies confirmed previous observations that Ku is undetectable in most nonprimate cells. However, small amounts of Ku could be detected in MOPC-315, but not L-929, cells by immunoprecipitating with mAb 162. In addition, autoantibodies to Ku were identified in the sera of approximately 1/3 of MRL/lpr mice. The murine autoantibodies also immunoprecipitated a small amount of Ku (comparable to that seen with 162) from MOPC-315, but not L-929, cell lysates. Characterization of the mAb specificities by immunoblot analysis with Ku fusion proteins revealed that mAbs 111, S10B1, and N9C1 bound to distinct epitopes of human p80 (amino acids 610-705, 8-221, and 1-374, respectively). All three mAbs were unreactive with murine p80. MAbs N3H10 and S5C11 bound immediately adjacent to the DNA binding site of p70 (amino acids 506-541). Only N3H10 displayed comparable reactivity with human and murine p70 on immunoblots, but it immunoprecipitated murine Ku poorly. S5C11 crossreacted more weakly with murine p70 on immunoblots, whereas 162 was completely unreactive with human or murine Ku on immunoblots, despite immunoprecipitating Ku efficiently. Studies with mAbs N3H10 and 162 suggest that the level of Ku is considerably lower in nonprimate cells than cells of primate origin, and that L-929 cells express little or no Ku protein.(ABSTRACT TRUNCATED AT 400 WORDS)

Authors
Wang, J; Chou, CH; Blankson, J; Satoh, M; Knuth, MW; Eisenberg, RA; Pisetsky, DS; Reeves, WH
MLA Citation
Wang, J, Chou, CH, Blankson, J, Satoh, M, Knuth, MW, Eisenberg, RA, Pisetsky, DS, and Reeves, WH. "Murine monoclonal antibodies specific for conserved and non-conserved antigenic determinants of the human and murine Ku autoantigens." Mol Biol Rep 18.1 (June 1993): 15-28.
PMID
7694076
Source
pubmed
Published In
Molecular Biology Reports
Volume
18
Issue
1
Publish Date
1993
Start Page
15
End Page
28

The influence of DNA structure on the in vitro stimulation of murine lymphocytes by natural and synthetic polynucleotide antigens.

Although DNA is generally considered to be a poor immunogen, recent evidence suggests that DNA from various species differ in their immunological activity and that bacterial DNA can induce the in vitro proliferation of normal murine B cells. To delineate structural features of DNA associated with mitogenic activity, the response of murine lymphocytes to various natural and synthetic polynucleotides was determined. Both ss and dsDNA from two different bacterial strains were equally effective in inducing proliferation. This response was independent of adenosine methylation, since DNA from dam- Escherichia coli stimulated proliferation. Among the synthetic polymers tested, only the duplexes poly(dG).poly(dC), and poly(dG.dC) were mitogenic, while polymers containing dA, dT, or dI alone or in combination with dG and dC were inactive. The mitogenic activity of poly(dG.dC) was eliminated, however, upon substitution of rG for dG or 5medC for dC. The mitogenic activity did not require high molecular weight DNA since active polymers ranged in size from approximately 260 to 800 base pairs. In addition, E. coli DNA fragments of 50-300 and 125-600 bases were mitogenic. Together, these data suggest that the mitogenic activity of DNA is dependent on sequence-specific determinants that can be presented by synthetic DNA duplexes as well as bacterial ss and dsDNA.

Authors
Messina, JP; Gilkeson, GS; Pisetsky, DS
MLA Citation
Messina, JP, Gilkeson, GS, and Pisetsky, DS. "The influence of DNA structure on the in vitro stimulation of murine lymphocytes by natural and synthetic polynucleotide antigens." Cell Immunol 147.1 (March 1993): 148-157.
PMID
8462107
Source
pubmed
Published In
Cellular Immunology
Volume
147
Issue
1
Publish Date
1993
Start Page
148
End Page
157
DOI
10.1006/cimm.1993.1055

V region gene analysis of anti-Sm hybridomas from MRL/Mp-lpr/lpr mice.

Anti-Sm autoantibodies are unique to SLE, but are present in only 25% of patients with this disease. This response also occurs at a similar frequency in mice of the autoimmune MRL strains. Previous analyses of the anti-Sm response in these mice indicate that its occurrence is controlled by stochastic events, and suggest that Sm is the driving Ag. To further elucidate the role of Ag in this response, and to test the hypothesis that the 25% incidence is due to a requirement for particular Ig gene rearrangements or somatic mutations, we have analyzed the specificity and V-region gene sequences of 41 anti-Sm B cell hybridomas derived from nine anti-Sm-positive MRL/Mp-lpr/lpr mice. The majority of hybridomas are specific for the D peptide of the Sm particle. Hybridomas of independent origin express unique VH/V kappa combinations with diverse junctional sequences and are variable in the extent of somatic mutation. Thus, the response does not appear to be dependent upon the occurrence of a rare Ig gene rearrangement or specific somatic mutation. The response exhibits restriction in JH and VH gene use, and in individual mice is oligoclonal, suggestive of Ag selection. In the few B cells for which mutations can be identified, the evidence for selection of mutant B lymphocytes, based on patterns of mutation, is ambiguous. However, there is remarkably little intraclonal diversity, suggesting that the overall mutation rates in these clones are low.

Authors
Bloom, DD; Davignon, JL; Retter, MW; Shlomchik, MJ; Pisetsky, DS; Cohen, PL; Eisenberg, RA; Clarke, SH
MLA Citation
Bloom, DD, Davignon, JL, Retter, MW, Shlomchik, MJ, Pisetsky, DS, Cohen, PL, Eisenberg, RA, and Clarke, SH. "V region gene analysis of anti-Sm hybridomas from MRL/Mp-lpr/lpr mice." J Immunol 150.4 (February 15, 1993): 1591-1610.
PMID
8432995
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
150
Issue
4
Publish Date
1993
Start Page
1591
End Page
1610

Characterization of glomerular thromboxane receptors in murine lupus nephritis.

Renal thromboxane (Tx) production is increased in the MRL-lpr murine model of lupus nephritis. To investigate the relationship between increased Tx production and number and affinity of Tx receptors, we measured binding of the Tx receptor antagonist [3H][SQ295481S-1 alpha,2 beta(5Z),3 beta,4 alpha]-7-(3-((2-((phenyl- amino)-carbonyl)hydrozino)methyl)-7-oxabicyclo-(2.2.1)heptan -2-yl)-5-heptenoic acid in glomerular preparations from MRL-lpr mice and both MRL(-)+/+ and LG/J controls. Renal Tx binding was first characterized in normal LG/J mice. In these animals, glomerular binding was specific, saturable and reversible. Scatchard analysis revealed a single class of high-affinity binding sites. We next evaluated Tx production and binding in 12- and 16-week-old MRL-lpr mice and MRL(-)+/+ controls. To assess renal Tx production, excretion of TxB2 was measured in urine. Urinary TxB2 was increased in MRL-lpr mice at 16 weeks of age. This increase in urinary TxB2 was associated with a reduction in density of glomerular Tx binding sites compared to either 12-week-old MRL-lpr mice or MRL(-)+/+ controls. Ligand binding affinity was similar in all groups. To investigate if this alteration in binding was specific for Tx, glomerular binding of [3H]angiotensin II was measured. In MRL-lpr mice, the number and affinity of glomerular angiotensin binding sites were similar at 12 and 16 weeks of age. Thus, in this murine model of lupus nephritis, enhanced renal Tx production is temporally associated with a decrease in glomerular Tx binding sites without a change in receptor affinity.(ABSTRACT TRUNCATED AT 250 WORDS)

Authors
Spurney, RF; Onorato, JJ; Ruiz, P; Pisetsky, DS; Coffman, TM
MLA Citation
Spurney, RF, Onorato, JJ, Ruiz, P, Pisetsky, DS, and Coffman, TM. "Characterization of glomerular thromboxane receptors in murine lupus nephritis." J Pharmacol Exp Ther 264.2 (February 1993): 584-590.
PMID
8437109
Source
pubmed
Published In
The Journal of pharmacology and experimental therapeutics
Volume
264
Issue
2
Publish Date
1993
Start Page
584
End Page
590

The fine specificity of monoclonal anti-DNA antibodies induced in normal mice by immunization with bacterial DNA.

To evaluate further bacterial DNA immunization as a model to study antigen drive in the anti-DNA response, the specificity of induced monoclonal anti-DNA antibodies was characterized. A panel of IgM and IgG monoclonal anti-DNA antibodies was produced from spleen cells of BALB/c mice immunized with single-stranded DNA from E. coli complexed to methylated bovine serum albumin in complete Freund's adjuvant. The binding of these antibodies to DNA and non-DNA antigens was tested by ELISA to assess their range of polyspecificity. These monoclonal antibodies were found to bind to nucleic acid as well as non-nucleic acid antigens, such as beta-galactosidase, cardiolipin, Ro, La and Sm. These studies demonstrate that anti-DNA antibodies from normal mice, although induced by bacterial DNA, may display a broad range of antigen recognition and thus resemble lupus anti-DNA antibodies, many of which are polyspecific, in their pattern of cross-reactivity.

Authors
Pyun, EH; Pisetsky, DS; Gilkeson, GS
MLA Citation
Pyun, EH, Pisetsky, DS, and Gilkeson, GS. "The fine specificity of monoclonal anti-DNA antibodies induced in normal mice by immunization with bacterial DNA." J Autoimmun 6.1 (February 1993): 11-26.
PMID
8457283
Source
pubmed
Published In
Journal of Autoimmunity
Volume
6
Issue
1
Publish Date
1993
Start Page
11
End Page
26
DOI
10.1006/jaut.1993.1002

Genetic analysis of MRL-lpr mice: relationship of the Fas apoptosis gene to disease manifestations and renal disease-modifying loci.

In MRL mice, the mostly recessive lpr mutation results in both the accumulation of CD4-, CD8-, CD3+ T cells in lymphoid tissue and many features of generalized autoimmune disease, including immune complex glomerulonephritis. To positionally clone the lpr mutation and analyze the effects of background genes, backcross offspring were examined from the cross: (MRL/MpJ-lpr x CAST/Ei)F1 x MRL/MpJ-lpr. The lpr gene was found to be closely linked to a mouse chromosome 19 marker defined by a variation of a Fas gene restriction fragment. Our results identified differences in RNA expression and differences in the genomic organization of the Fas gene between normal and lpr mice, and confirm the recent report that a mutation in the Fas apoptosis gene is the lpr mutation. However, our results also indicate that the Fas gene is expressed in spleen cells from normal mice, and spleen and lymph node cells from mice with a second mutation at the lpr locus (lprcg). Together these results suggest that altered Fas transcription results in the failure of lymphocytes to undergo programmed cell death and may lead to an altered immune cell repertoire. This mechanism may explain certain central and peripheral defects in tolerance that are present in autoimmune disease. The current study also demonstrates the profound effect of background genes on the degree of nephritis, lymphadenopathy, and anti-DNA antibody production. Of major note, our studies suggest the identification of chromosomal positions for genes that modify nephritis. Analysis of the backcross mice for markers covering most of the mouse genome suggests that over 50% of the variance in renal disease is attributable to quantitative trait loci on mouse chromosomes 7 and 12. Moreover, this study provides a model for dissecting the complex genetic interactions that result in manifestations of autoimmune disease.

Authors
Watson, ML; Rao, JK; Gilkeson, GS; Ruiz, P; Eicher, EM; Pisetsky, DS; Matsuzawa, A; Rochelle, JM; Seldin, MF
MLA Citation
Watson, ML, Rao, JK, Gilkeson, GS, Ruiz, P, Eicher, EM, Pisetsky, DS, Matsuzawa, A, Rochelle, JM, and Seldin, MF. "Genetic analysis of MRL-lpr mice: relationship of the Fas apoptosis gene to disease manifestations and renal disease-modifying loci." J Exp Med 176.6 (December 1, 1992): 1645-1656.
PMID
1460423
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
176
Issue
6
Publish Date
1992
Start Page
1645
End Page
1656

Specificity analysis of antibodies to single-stranded micrococcal DNA in the sera of normal human subjects and patients with systemic lupus erythematosus.

To evaluate the properties of antibodies to bacterial DNA in the sera of normal human subjects (NHS) and patients with systemic lupus erythematosus (SLE), the effects of ionic strength and pH on their binding to single-stranded DNA (ssDNA) from Micrococcus lysodeikticus (MC) were measured. By ELISA, antibodies to MC ssDNA in NHS showed greater activity at high ionic strength (0.2-1.0 M) than antibodies in lupus sera. Similarly, antibodies in NHS had higher activity at pH 9 than lupus anti-DNA. Competition binding assays indicated, moreover, that NHS anti-DNA showed greater inhibition by DNA than lupus anti-DNA at comparable inhibitor concentrations. Together, these results suggest that antibodies to MC ssDNA in NHS and SLE sera may differ in their mode of interaction with bacterial DNA and that NHS can generate high avidity antibodies to at least certain DNA determinants.

Authors
Robertson, CR; Pisetsky, DS
MLA Citation
Robertson, CR, and Pisetsky, DS. "Specificity analysis of antibodies to single-stranded micrococcal DNA in the sera of normal human subjects and patients with systemic lupus erythematosus." Clin Exp Rheumatol 10.6 (November 1992): 589-594.
PMID
1483310
Source
pubmed
Published In
Clinical and experimental rheumatology
Volume
10
Issue
6
Publish Date
1992
Start Page
589
End Page
594

Autoantibodies and their idiotypes.

Antinuclear antibodies occur prominently in systemic lupus erythematosus and serve as markers of underlying pathogenetic disturbances. Although these antibodies display features indicative of genetic control and in vivo selection by self-antigen, other mechanisms shaping the B-cell repertoire may influence their production. Provocative new animal models provide systems for analyzing the cellular and genetic disturbances promoting these responses, as well as the role of pathogenic specificities in inducing tissue injury.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Autoantibodies and their idiotypes." Curr Opin Rheumatol 4.5 (October 1992): 635-643. (Review)
PMID
1419497
Source
pubmed
Published In
Current Opinion in Rheumatology
Volume
4
Issue
5
Publish Date
1992
Start Page
635
End Page
643

Effect of anti-CD4 antibody treatment on inflammatory arthritis in MRL-lpr/lpr mice.

MRL-lpr/lpr mice develop an inflammatory arthritis in association with other manifestations of autoimmunity. Although a variety of immune cell disturbances have been described in these mice, the relationship of these abnormalities to the pathogenesis of arthritis has not yet been determined; the role of T cells is especially unclear since synovial hypertrophy and joint erosions have been noted in some studies in the absence of a significant T cell infiltrate. Therefore, to determine if T cells are required for arthritis in MRL-lpr/lpr mice, we evaluated the effects of prolonged treatment with a monoclonal anti-CD4 antibody. Knee joints from treated mice had markedly reduced arthritis compared to saline-treated control animals as measured by the degree of synovial hypertrophy and inflammation. Nephritis in these mice was concomitantly reduced. In contrast, rheumatoid factor levels were not affected by CD4+ cell depletion, despite significant effects on anti-DNA. These results indicate that in MRL-lpr/lpr mice anti-CD4 therapy can inhibit arthritis, suggesting an important role of T cells in the pathogenesis of this lesion.

Authors
Gilkeson, GS; Spurney, R; Coffman, TM; Kurlander, R; Ruiz, P; Pisetsky, DS
MLA Citation
Gilkeson, GS, Spurney, R, Coffman, TM, Kurlander, R, Ruiz, P, and Pisetsky, DS. "Effect of anti-CD4 antibody treatment on inflammatory arthritis in MRL-lpr/lpr mice." Clin Immunol Immunopathol 64.2 (August 1992): 166-172.
PMID
1353712
Source
pubmed
Published In
Clinical Immunology and Immunopathology
Volume
64
Issue
2
Publish Date
1992
Start Page
166
End Page
172

A defect in the humoral immune response to protein antigens and haptens in immunoglobulin mu heavy-chain transgenic mice.

We have examined the antibody response in mice expressing a functionally rearranged mu Ig heavy chain derived from a hybridoma antibody with specificity for the hapten 4-hydroxy-3-nitrophenyl (NP). Transgenic mice and their normal littermates were immunized with the antigens NP-OVA, the synthetic polypeptide (Tyr,Glu)-Ala-Lys ((T,G)-A-L), or saline. The presence of serum antibodies to NP-BSA, OVA, (T,G)-A-L, and BSA was examined by ELISA. Sera were evaluated prior to immunization and at periods of up to 4 months following immunization. Prior to immunization, transgenic mice had high levels of IgM anti-NP antibody but no detectable antibody to the other antigens. Both the primary and secondary antibody responses of transgenic mice to NP, OVA, and (T,G)-A-L were depressed when compared with the response of non-transgenic mice. Because of reports that these transgenic mice have increased proportions of CD5 + B-cells, a subpopulation associated with the production of autoantibodies, we examined these mice for the production of both IgG and IgM rheumatoid factors and anti-DNA antibodies. Transgenic mice had a modest increase in the spontaneous production of IgM anti-DNA. These data demonstrate a functional defect in the humoral immune response of mu transgenic mice.

Authors
Pincus, SH; Cole, R; Pisetsky, DS
MLA Citation
Pincus, SH, Cole, R, and Pisetsky, DS. "A defect in the humoral immune response to protein antigens and haptens in immunoglobulin mu heavy-chain transgenic mice." Mol Immunol 29.6 (June 1992): 801-806.
PMID
1603097
Source
pubmed
Published In
Molecular Immunology
Volume
29
Issue
6
Publish Date
1992
Start Page
801
End Page
806

Anti-DNA antibodies in systemic lupus erythematosus.

Anti-DNA antibodies are the serologic hallmark of systemic lupus erythematosus and important markers for diagnosis and prognosis. Although a number of mechanisms for anti-DNA production have been proposed, recent evidence from human as well as murine lupus indicate a prominent role of DNA antigen drive. To identify further the basis of this response, current research is focusing on the structural features of DNA associated with immunogenicity in both the normal and autoimmune settings. Elucidating these mechanisms should thus provide insights into the pathogenesis of systemic lupus erythematosus as well as the immunologic activity of nucleic acids.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Anti-DNA antibodies in systemic lupus erythematosus." Rheum Dis Clin North Am 18.2 (May 1992): 437-454. (Review)
PMID
1626077
Source
pubmed
Published In
Rheumatic Disease Clinics of North America
Volume
18
Issue
2
Publish Date
1992
Start Page
437
End Page
454

Thromboxane receptor blockade reduces renal injury in murine lupus nephritis.

To investigate the role of thromboxane A2 (TxA2) in murine lupus, we assessed the effects of the specific thromboxane receptor antagonist GR32191 on immune complex glomerulonephritis in MRL-lpr/lpr mice. Forty mg/kg/day GR32191 was given by twice daily subcutaneous injection for eight weeks beginning at 12 weeks of age. This dose completely blocked the renal vasoconstriction produced by the thromboxane agonist U46619. After eight weeks of treatment, both glomerular filtration rate (GFR) (8.9 +/- 0.6 vs. 6.8 +/- 1.1 ml/min/kg; P less than 0.05) and PAH clearance (CPAH) (37.4 +/- 2.5 vs. 29.9 +/- 3.3 ml/min/kg; P less than 0.05) were significantly higher in mice given GR32191 compared to vehicle treated animals. Administration of GR32191 also reduced proteinuria from 18.1 +/- 11.6 to 3.7 +/- 1.3 mg/24 hours (P less than 0.05). In GR32191 treated MRL-lpr/lpr mice, renal hemodynamic function and proteinuria were not significantly different from congenic MRL-+/+ controls. Thromboxane receptor blockade had striking affects on renal histomorphology reducing both hyaline thrombi in glomeruli (P = 0.022) and interstitial inflammation (P = 0.006). Glomerular crescents and severity of vasculitis also tended to be reduced in mice receiving the thromboxane receptor antagonist. The overall histopathologic score in mice given GR32191 was significantly lower than vehicle treated animals (4.7 +/- 0.5 vs. 8.4 +/- 1.5; P = 0.016). These effects of GR32191 were associated with decreased excretion of thromboxane B2 (TxB2) in urine (292 +/- 37 vs. 747 +/- 155 pg/24 hr; P less than 0.005) as well as a modest reduction in glomerular deposits of IgG (semiquantitative score 2.6 +/- 0.2 vs. 3.5 +/- 0.2; P less than 0.02). Thus, chronic thromboxane receptor blockade markedly altered the course of renal disease in MRL-lpr/lpr mice, suggesting that TxA2 is an important mediator of renal dysfunction and injury in this murine model of lupus nephritis.

Authors
Spurney, RF; Fan, PY; Ruiz, P; Sanfilippo, F; Pisetsky, DS; Coffman, TM
MLA Citation
Spurney, RF, Fan, PY, Ruiz, P, Sanfilippo, F, Pisetsky, DS, and Coffman, TM. "Thromboxane receptor blockade reduces renal injury in murine lupus nephritis." Kidney Int 41.4 (April 1992): 973-982.
PMID
1387435
Source
pubmed
Published In
Kidney international
Volume
41
Issue
4
Publish Date
1992
Start Page
973
End Page
982

Analysis of the expression of CD5 by human B cells and correlation with functional activity.

B cells expressing the CD5 marker in the mouse have been suggested to be a separate lineage and a major source of autoantibody production. In man, this relationship is less clear. Studies were therefore undertaken to determine whether human CD5+ B cells represent a distinct lineage of cells that differ in patterns of antibody production from CD5- B cells. In normal B cell populations, CD5 was expressed by a mean of 24.0 +/- 2.8% (n = 10) of CD20+ B cells. Of note, an increased frequency of CD5+ B cells was not found in patients with systemic lupus erythematosus (mean of 17.9 +/- 2.8%, n = 16). Analyzing CD5+ B cells for cell membrane Ig isotype expression demonstrated similar frequencies of IgG and IgA expressing cells as were found on the CD5- B cell population, although the frequency of IgM+ cells was slightly increased. Incubation of CD20+ B cells with phorbol myristate acetate (PMA) for 72 hr increased the frequency of CD5 expressing B cells by more than threefold. CD5 expression was also increased by coculture with anti-CD3-activated T cells and most markedly by simultaneous stimulation with both PMA- and anti-CD3-activated T cells (greater than 50% positive). Analysis of CD5- B cells clearly indicated that stimulation with PMA or anti-CD3-activated T cells induced the majority to become CD5+ transiently. Functional analysis of Ig production by CD5+ and CD5- B cells stimulated with anti-CD3-activated T cells indicated that both populations in normals produced IgM and a variety of autoantibodies in comparable amounts, whereas the CD5- B cells produced greater quantities of IgG. B cells were activated with anti-CD3-stimulated T cells followed by separation into CD5+ and CD5- populations. The largest amount of Ig was produced by CD5- B cells that were induced to express CD5, although all populations produced some Ig. These data suggest that CD5 behaves as an activation marker on human B cells rather than as a marker for a distinct lineage of cells. Moreover, CD5 expression does not appear to identify a population of resting B cells with a greater capacity to produce antibodies to DNA or other autoantibodies.

Authors
Vernino, LA; Pisetsky, DS; Lipsky, PE
MLA Citation
Vernino, LA, Pisetsky, DS, and Lipsky, PE. "Analysis of the expression of CD5 by human B cells and correlation with functional activity." Cell Immunol 139.1 (January 1992): 185-197.
PMID
1370255
Source
pubmed
Published In
Cellular Immunology
Volume
139
Issue
1
Publish Date
1992
Start Page
185
End Page
197

Patterns of heavy and light chain utilization in the antibody response to single-stranded bacterial DNA in normal human subjects and patients with systemic lupus erythematosus.

Although anti-DNA antibodies are generally considered to be specific markers for systemic lupus erythematosus (SLE), antibodies binding DNA from certain bacterial species can be found in the sera of normal subjects. To characterize the immunochemical properties of these antibodies, the IgG subclass and light chain profile of antibodies to single-stranded micrococcal DNA (MC DNA) in the sera of normal subjects and patients with SLE was determined. The anti-MC DNA response in normal sera was predominantly of the IgG2 subclass with a marked predominance of kappa light chains. In contrast, anti-MC DNA antibodies in SLE sera exhibited all IgG subclasses with a predominance of the IgG1 subclass and both kappa and lambda light chains were represented. These results suggest that antibodies to bacterial DNA in the sera of normal subjects and patients with SLE differ in patterns of immunoglobulin gene expression; the restricted response of normal subjects may be related to the binding to a discrete DNA determinant.

Authors
Robertson, CR; Gilkeson, GS; Ward, MM; Pisetsky, DS
MLA Citation
Robertson, CR, Gilkeson, GS, Ward, MM, and Pisetsky, DS. "Patterns of heavy and light chain utilization in the antibody response to single-stranded bacterial DNA in normal human subjects and patients with systemic lupus erythematosus." Clin Immunol Immunopathol 62.1 Pt 1 (January 1992): 25-32.
PMID
1728977
Source
pubmed
Published In
Clinical Immunology and Immunopathology
Volume
62
Issue
1 Pt 1
Publish Date
1992
Start Page
25
End Page
32

Mechanisms of anti-DNA antibody expression in normal and aberrant immunity.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Mechanisms of anti-DNA antibody expression in normal and aberrant immunity." Concepts Immunopathol 8 (1992): 71-84. (Review)
PMID
1735106
Source
pubmed
Published In
Concepts in immunopathology
Volume
8
Publish Date
1992
Start Page
71
End Page
84

Immunochemical properties of anti-DNA antibodies in the sera of patients with Escherichia coli bacteremia.

To assess the role of infection in anti-DNA antibody production, the DNA-binding activity of sera from patients with Escherichia coli bacteremia was analyzed. Among 8 patients with bacteremia documented by blood culture, 5 demonstrated increased levels of antibodies to single-stranded DNA from E. coli as measured by enzyme-linked immunosorbent assay. Sera from these patients also reacted with single-stranded DNA from other bacterial and mammalian species as well as certain synthetic polynucleotides including poly-dT and poly-dC. The isotype distribution of these antibodies and their avidity as assessed by competition enzyme-linked immunosorbent assay resembled, moreover, responses of patients with systemic lupus erythematosus. These results suggest that, during the course of infection with E. coli, some patients may produce antibodies with immunochemical properties similar to those arising in systemic lupus erythematosus.

Authors
Robertson, CR; Pisetsky, DS
MLA Citation
Robertson, CR, and Pisetsky, DS. "Immunochemical properties of anti-DNA antibodies in the sera of patients with Escherichia coli bacteremia." Int Arch Allergy Immunol 98.4 (1992): 311-316.
PMID
1422260
Source
pubmed
Published In
International archives of allergy and immunology
Volume
98
Issue
4
Publish Date
1992
Start Page
311
End Page
316

Patterns of heavy and light chain utilization in the antibody response to single-stranded bacterial DNA in normal human subjects and patients with systemic lupus erythematosus

Although anti-DNA antibodies are generally considered to be specific markers for systemic lupus erythematosus (SLE), antibodies binding DNA from certain bacterial species can be found in the sera of normal subjects. To characterize the immunochemical properties of these antibodies, the IgG subclass and light chain profile of antibodies to single-stranded micrococcal DNA (MC DNA) in the sera of normal subjects and patients with SLE was determined. The anti-MC DNA response in normal sera was predominantly of the IgG2 subclass with a marked predominance of κ light chains. In contrast, anti-MC DNA antibodies in SLE sera exhibited all IgG subclasses with a predominance of the IgG1 subclass and both κ and λ light chains were represented. These results suggest that antibodies to bacterial DNA in the sera of normal subjects and patients with SLE differ in patterns of immunoglobulin gene expression; the restricted response of normal subjects may be related to the binding to a discrete DNA determinant. © 1992.

Authors
Robertson, CR; Gilkeson, GS; Ward, MM; Pisetsky, DS
MLA Citation
Robertson, CR, Gilkeson, GS, Ward, MM, and Pisetsky, DS. "Patterns of heavy and light chain utilization in the antibody response to single-stranded bacterial DNA in normal human subjects and patients with systemic lupus erythematosus." Clinical Immunology and Immunopathology 62.1 PART 1 (1992): 25-32.
Source
scival
Published In
Clinical Immunology and Immunopathology
Volume
62
Issue
1 PART 1
Publish Date
1992
Start Page
25
End Page
32
DOI
10.1016/0090-1229(92)90019-K

Systemic lupus erythematosus.

Systemic lupus erythematosus is a prototypic autoimmune disease characterized by antinuclear antibody production. In recent investigations, the contributions of various polymorphic immune response gene systems to disease pathogenesis have been analyzed. Unique cellular and molecular studies have also established the role of 'autoantigen drive' in autoantibody induction and its relationship to polyclonal B-cell activation.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Systemic lupus erythematosus." Curr Opin Immunol 3.6 (December 1991): 917-923. (Review)
PMID
1793536
Source
pubmed
Published In
Current Opinion in Immunology
Volume
3
Issue
6
Publish Date
1991
Start Page
917
End Page
923

Autoantibodies and their idiotypes.

Idiotypes are serologically defined markers in the variable region of an antibody molecule. In the study of autoimmunity, these markers have been valuable probes in defining patterns of autoantibody variable region gene utilization, mechanisms of immune dysregulation, and the operation of the network in the generation of abnormal responses. Recent advances in the molecular analysis of immunoglobulin genes have provided insights into the structural basis of autoantibody idiotypes and the manner in which sequence homologies between idiotype-bearing antibodies and autoantigens can lead to autoreactivity. Immune manipulation using these sequences could lead to novel models of disease as well as prospects for immunotherapy.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Autoantibodies and their idiotypes." Curr Opin Rheumatol 3.5 (October 1991): 731-737. (Review)
PMID
1751308
Source
pubmed
Published In
Current Opinion in Rheumatology
Volume
3
Issue
5
Publish Date
1991
Start Page
731
End Page
737

Stimulation of in vitro murine lymphocyte proliferation by bacterial DNA.

Although DNA is generally considered to be a poor immunogen, recent evidence suggests that DNA from various species differ in their immunologic activity and that bacterial DNA, unlike mammalian DNA, can induce significant antibody responses in mice. To explore further the immunologic activities of bacterial DNA, its ability to stimulate in vitro proliferation of murine lymphocytes was tested. The stimulation of lymphocytes with highly purified ssDNA from Escherichia coli resulted in a dose-dependent response that was maximal at 48 h. Several lines of evidence indicate that DNA, rather than endotoxin contamination, induced this response: 1) LPS at doses equivalent to those detected in the DNA preparation caused significantly less proliferation than the DNA; 2) the response to DNA was insensitive to polymyxin B; 3) pretreatment of DNA with DNase completely abrogated the response; and 4) DNA induced the proliferation of cells from endotoxin-resistant C3H/HeJ mice. Furthermore, although DNA from three different bacterial species induced proliferation, mammalian DNA from three species were nonmitogenic. Depletion of T cells from lymphocytes did not reduce proliferation, suggesting that bacterial DNA directly triggered B cell proliferation. These studies provide further evidence that DNA are not uniform in their immunologic activities likely because of their content of nonconserved structural determinants.

Authors
Messina, JP; Gilkeson, GS; Pisetsky, DS
MLA Citation
Messina, JP, Gilkeson, GS, and Pisetsky, DS. "Stimulation of in vitro murine lymphocyte proliferation by bacterial DNA." J Immunol 147.6 (September 15, 1991): 1759-1764.
PMID
1890302
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
147
Issue
6
Publish Date
1991
Start Page
1759
End Page
1764

Soluble interleukin-2 receptor levels in patients with dermatitis herpetiformis.

To determine the role of T-cell activation in dermatitis herpetiformis (DH), soluble IL-2R levels were measured by enzyme-linked immunosorbent assay (ELISA) in the sera of 30 patients with DH. Levels of this shed receptor are considered to be a measure of in vivo T-lymphocyte activation, and are elevated in the sera of many patients with inflammatory and immune-mediated diseases. Fifteen of the thirty (50%) patients with DH had elevated levels of soluble IL-2R compared to one of 31 (3%) healthy HLA-B8 or HLA-DR3 control subjects (p less than 0.00001) and one of 10 (10%) healthy non-HLA-B8/-DR3 subjects (p less than 0.0018). In addition, the mean soluble IL-2R level in the patients with DH (744 +/- 381 U/ml) was also significantly higher than that seen in 31 healthy HLA B8 or HLA DR3 individuals (388 +/- 160 U/ml, p = 0.0001) and 10 healthy non-HLA-B8/DR3 individuals (397 +/- 201 U/ml, p = 0.002). Only two of the 30 patients with DH had active skin lesions at the time of serum sampling, one of whom had elevated levels of IL-2R. Measurement of soluble IL-2R levels in sequential serum samples, available in four patients with DH at times of active and inactive skin disease, demonstrated a temporal association between soluble IL-2R level elevations and active skin disease in two patients and no association in two patients. In one patient a marked elevation in soluble IL-2R levels occurred with the onset of gastrointestinal symptoms, which decreased by 14% with institution of a gluten-free diet. In order to determine if soluble IL-2R levels are related to the mucosal immune response, the IL-2R levels were compared to the level of IgA antibodies directed against the dietary antigen beta-lactoglobulin. Ten of eleven (91%) patients with circulating IgA anti-beta lactoglobulin antibodies were also found to have elevated levels of IL-2R. In contrast, in the patients with no detectable IgA anti-beta lactoglobulin antibodies, only four of 16 (25%) had elevated levels of IL-2R (p = 0.001). Because IL-2R levels are not related to activity of the skin disease in patients with DH but are associated with the presence of IgA antibodies against the dietary antigen beta-lactoglobulin, these results suggest that some of the T-cell activation commonly present in DH reflects an ongoing immune response in the gastrointestinal tract.

Authors
Ward, MM; Pisetsky, DS; Hall, RP
MLA Citation
Ward, MM, Pisetsky, DS, and Hall, RP. "Soluble interleukin-2 receptor levels in patients with dermatitis herpetiformis." J Invest Dermatol 97.3 (September 1991): 568-572.
PMID
1875055
Source
pubmed
Published In
Journal of Investigative Dermatology
Volume
97
Issue
3
Publish Date
1991
Start Page
568
End Page
572

Genetic control of inflammatory arthritis and glomerulonephritis in congenic lpr mice and their F1 hybrids.

MRL-lpr/lpr mice spontaneously develop a complex immunological disease characterized by glomerulonephritis, inflammatory erosive arthritis and the production of rheumatoid factors (RF) and anti-DNA antibodies. We have previously reported that, of congenic lpr strains, only MRL-lpr/lpr mice develop synovial pathology suggesting that both the lpr gene and another gene(s) in the MRL background are necessary for the development of arthritis. To define further the genetics of arthritis and its relationship to glomerulonephritis and autoantibody production, we studied disease expression in MRL-lpr/lpr and C57BL/6-lpr/lpr mice and their offspring (BM-lpr/lpr and MB-lpr/lpr). At 6 months of age these mice were killed and bled, and their kidneys and knee joints were removed for pathological studies. Fourteen of 28 MB-lpr/lpr mice displayed synovial hypertrophy, while eight of 28 had significant synovial inflammation. BM-lpr/lpr mice showed similar changes: nine of 22 and eight of 22 exhibited synovial hypertrophy and inflammation respectively. Joints from MRL-lpr/lpr mice revealed 13 of 17 with synovial hypertrophy and 12 of 17 with inflammation, while none of 14 B6-lpr/lpr mice had synovial changes. Renal pathology was minimal in the F1 mice with only mild hypercellularity in seven of 21 MB-lpr/lpr and five of 22 BM-lpr/lpr mice. All MRL-lpr/lpr mice, in contrast, had marked glomerular changes with 12 of 17 exhibiting glomerular crescents. Only one F1 mouse had both arthritis and renal abnormalities. IgM RF levels were elevated in all four experimental groups, but did not correlate with the presence or severity of arthritis. IgG RF levels were elevated in the MB-lpr/lpr and MRL-lpr/lpr mice, but did not correlate with the degree of arthritis. These results indicate that renal disease and arthritis develop independently in lpr mice, possibly on a genetic basis, and that the presence and titer of autoantibodies do not correlate with tissue injury.

Authors
Gilkeson, GS; Ruiz, P; Pritchard, AJ; Pisetsky, DS
MLA Citation
Gilkeson, GS, Ruiz, P, Pritchard, AJ, and Pisetsky, DS. "Genetic control of inflammatory arthritis and glomerulonephritis in congenic lpr mice and their F1 hybrids." J Autoimmun 4.4 (August 1991): 595-606.
PMID
1777011
Source
pubmed
Published In
Journal of Autoimmunity
Volume
4
Issue
4
Publish Date
1991
Start Page
595
End Page
606

Physiologic role for enhanced renal thromboxane production in murine lupus nephritis.

To investigate the physiologic significance of enhanced renal thromboxane production in murine lupus nephritis, we measured renal hemodynamics and eicosanoid production in MRL-lpr/lpr mice from 8 to 20 weeks of age. Over this age range, MRL-lpr/lpr mice develop an autoimmune disease with nephritis similar to human systemic lupus erythematosus (SLE). In these studies, glomerular filtration rate (GFR) and PAH clearance (CPAH) decreased progressively with age in MRL-lpr/lpr mice, but not in controls. This impairment of renal hemodynamics was associated with increased renal thromboxane production, as well as increased excretion of both thromboxane B2 (TxB2) and 2,3-dinor TxB2 in urine. There was an inverse correlation between renal thromboxane production in MRL-lpr/lpr mice and both GFR and CPAH. Furthermore, there were positive correlations between thromboxane production by the kidney and both the severity of renal histopathology and serum anti-DNA antibody levels measured in individual animals. Enhanced urinary excretion of TxB2 and the development of renal dysfunction also coincided temporally with the appearance of increased levels of interleukin 1 beta (IL-1 beta) mRNA in renal cortex. Acute administration of the specific thromboxane receptor antagonist GR32191 to MRL-lpr/lpr mice restored GFR to normal in early stages of the autoimmune disease. However, in animals with more advanced nephritis, the effect of acute thromboxane receptor blockade on renal hemodynamics was less marked. We conclude that thromboxane A2 is an important mediator of reversible renal hemodynamic impairment in murine lupus, especially in the early phase of disease.

Authors
Spurney, RF; Bernstein, RJ; Ruiz, P; Pisetsky, DS; Coffman, TM
MLA Citation
Spurney, RF, Bernstein, RJ, Ruiz, P, Pisetsky, DS, and Coffman, TM. "Physiologic role for enhanced renal thromboxane production in murine lupus nephritis." Prostaglandins 42.1 (July 1991): 15-28.
PMID
1771236
Source
pubmed
Published In
Prostaglandins
Volume
42
Issue
1
Publish Date
1991
Start Page
15
End Page
28

The specialized centers of research in rheumatoid arthritis. Recent progress and prospects for future advances.

Specialized Centers of Research (SCOR) in arthritis are interdisciplinary research programs to investigate disease pathogenesis as well as advance diagnosis and treatment. A recent meeting of investigators from the three SCOR programs in rheumatoid arthritis demonstrated progress in several important research areas. Because of the multiplier effects of SCOR programs, new investigators have been enlisted into arthritis research as issues related to this disease become a focus of investigation throughout universities and medical centers. Continued progress by the SCOR programs should provide new targets for therapeutic intervention as well as strategies for monitoring disease activity.

Authors
Pisetsky, DS; Haynes, BF; Lipsky, PE; Kang, AH; Postlethwaite, AE
MLA Citation
Pisetsky, DS, Haynes, BF, Lipsky, PE, Kang, AH, and Postlethwaite, AE. "The specialized centers of research in rheumatoid arthritis. Recent progress and prospects for future advances." Hum Immunol 31.2 (June 1991): 148-151.
PMID
2066274
Source
pubmed
Published In
Human Immunology
Volume
31
Issue
2
Publish Date
1991
Start Page
148
End Page
151

Specificity of anti-DNA antibodies induced in normal mice by immunization with bacterial DNA.

To determine the specificity of anti-DNA antibodies induced in normal mice by immunization with bacterial DNA, sera from BALB/c mice immunized with single-stranded DNA from Escherichia coli (EC) were tested for binding to a panel of synthetic DNA and RNA homopolymers as well as duplexes. Results of these studies indicate that sera from EC DNA immunized mice preferentially bind certain DNA and RNA homopolymers as well as DNA duplexes. Furthermore, the specificity of the antibodies from immunized mice resembled those of sera from autoimmune MRL-lpr/lpr mice in terms of the synthetic antigens recognized, although some differences were noted in the magnitude of the response to individual duplexes. These results suggest that anti-DNA antibodies induced by bacterial DNA bind to DNA structures dependent on both the base and the sugar phosphate moieties of the nucleic acid antigen and may resemble some anti-DNA antibodies expressed in spontaneous autoimmune disease in these binding properties.

Authors
Gilkeson, GS; Pritchard, AJ; Pisetsky, DS
MLA Citation
Gilkeson, GS, Pritchard, AJ, and Pisetsky, DS. "Specificity of anti-DNA antibodies induced in normal mice by immunization with bacterial DNA." Clin Immunol Immunopathol 59.2 (May 1991): 288-300.
PMID
1706971
Source
pubmed
Published In
Clinical Immunology and Immunopathology
Volume
59
Issue
2
Publish Date
1991
Start Page
288
End Page
300

Serum immunoglobulin levels in systemic lupus erythematosus: the effects of age, sex, race and disease duration.

To determine whether factors other than disease activity influence immunoglobulin levels in patients with systemic lupus erythematosus (SLE), the effect of age, sex, race, and duration of disease on serum IgG and IgM levels in 170 patients with SLE were investigated. Serum IgM and IgG levels did not differ between men and women, while IgM levels were higher in whites. Serum IgG levels did not vary with age or duration of SLE. In contrast, serum IgM levels were negatively correlated with both age (r = -0.236; p = 0.002) and duration of SLE (r = 0.248; p = 0.001), and demonstrated a U-shaped age relationship, being higher in children and older patients. These patterns of immunoglobulin expression in patients with SLE contrast with those exhibited in populations of healthy individuals, suggesting that the immunoregulatory disturbances of SLE predominate over the normal mechanisms regulating levels of IgM and IgG.

Authors
Ward, MM; Dawson, DV; Pisetsky, DS
MLA Citation
Ward, MM, Dawson, DV, and Pisetsky, DS. "Serum immunoglobulin levels in systemic lupus erythematosus: the effects of age, sex, race and disease duration." J Rheumatol 18.4 (April 1991): 540-544.
PMID
2066946
Source
pubmed
Published In
The Journal of rheumatology
Volume
18
Issue
4
Publish Date
1991
Start Page
540
End Page
544

Anti-La antibody production by MRL-1pr/1pr mice. Analysis of fine specificity.

In evaluating the origin of autoantibodies, patterns of self-Ag recognition have been interpreted to reflect the relative role of Ag in stimulating a response. Few studies, however, have assessed whether human autoantibodies display patterns of autoantigen recognition similar to those of SLE-prone mice. In previous studies, anti-La antibodies from humans have been shown to bind multiple epitopes on recombinant human La Ag, including immunoreactivity with a large fragment, termed La C, representing the middle portion of the La sequence. We report herein for the first time that MRL-1pr mice also spontaneously produce antibodies to recombinant human La protein and resemble human autoantibodies in their reactivity with La C. To further investigate the fine specificity of this response, we tested for antibody binding to six synthetic La peptides representing sequences within La C. Whereas two of the synthetic La peptides reacted with MRL-1pr sera containing anti-La binding, low reactivity was observed with a large panel of human anti-La sera. Our results therefore show that patterns of La antigen recognition displayed by MRL-1pr antibodies differ from those of human autoantibodies, possibly reflecting differences between mouse and man in the induction of these responses.

Authors
St Clair, EW; Kenan, D; Burch, JA; Keene, JD; Pisetsky, DS
MLA Citation
St Clair, EW, Kenan, D, Burch, JA, Keene, JD, and Pisetsky, DS. "Anti-La antibody production by MRL-1pr/1pr mice. Analysis of fine specificity." J Immunol 146.6 (March 15, 1991): 1885-1892.
PMID
2005384
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
146
Issue
6
Publish Date
1991
Start Page
1885
End Page
1892

The relationship between soluble interleukin 2 receptor levels and antidouble stranded DNA antibody levels in patients with systemic lupus erythematosus.

Levels of soluble interleukin 2 receptors (IL-2R) have been found to be elevated in the serum of patients with systemic lupus erythematosus (SLE) and are considered an indication of immune system activation in this disease. To assess the relationship between soluble IL-2R levels and other serological markers, we compared levels of soluble IL-2R and anti-double stranded DNA (anti-dsDNA) antibodies in SLE sera. In a cross sectional study of 34 patients with SLE, soluble IL-2R levels were elevated compared with healthy individuals (1126 U/ml vs 235 U/ml, p less than 0.0001), and a significant correlation between levels of soluble IL-2R and anti-dsDNA antibodies was present (r = 0.543, p = 0.0009). In a longitudinal study of 10 additional patients, the time courses of soluble IL-2R levels and anti-dsDNA antibody measurements were similar in 3 patients. In 7 patients, substantial differences in the levels of soluble IL-2R and anti-dsDNA antibodies over time were observed, including marked changes in anti-dsDNA antibody levels that were accompanied by only minor changes in soluble IL-2R levels, and soluble IL-2R measurements that were persistently elevated despite generally low anti-dsDNA antibody levels. Our results indicate that soluble IL-2R levels may not always parallel other serological markers of SLE, suggesting measurement of different facets of immune system activation by various assays.

Authors
Ward, MM; Dooley, MA; Christenson, VD; Pisetsky, DS
MLA Citation
Ward, MM, Dooley, MA, Christenson, VD, and Pisetsky, DS. "The relationship between soluble interleukin 2 receptor levels and antidouble stranded DNA antibody levels in patients with systemic lupus erythematosus." J Rheumatol 18.2 (February 1991): 235-240.
PMID
2023217
Source
pubmed
Published In
The Journal of rheumatology
Volume
18
Issue
2
Publish Date
1991
Start Page
235
End Page
240

Enhanced renal leukotriene production in murine lupus: role of lipoxygenase metabolites.

To investigate the potential role of leukotrienes in murine lupus, we measured renal hemodynamics and renal leukotriene production in MRL-lpr/lpr mice at 12 and 20 weeks of age. Over this age range, these animals develop overt manifestations of autoimmune disease with nephritis similar to human SLE. In the current study, we demonstrated that glomerular filtration rate (GFR) and PAH clearance (CPAH) deteriorated with age in MRL-lpr/lpr mice, but not in MRL(-)+/+ controls. Impaired renal hemodynamic function in MRL-lpr/lpr mice was associated with enhanced ionophore-stimulated production of both leukotriene B4 (LTB4) and leukotriene C4 (LTC4) by preparations of renal cortex. There was a significant inverse correlation between GFR and in vitro production of both LTC4 and LTB4 in kidneys from MRL-lpr/lpr mice, but not in control animals. In addition, in vitro LTC4 production was correlated with the severity of renal histomorphologic abnormalities. Administration of the specific peptidoleukotriene receptor antagonist SKF104353 to 20 week old MRL-lpr/lpr mice significantly improved both GFR and CPAH, whereas this agent had no effect of renal hemodynamics in MRL(-)+/+ controls. These results suggest that renal production of LTC4 and LTB4 is increased in MRL-lpr/lpr mice with nephritis, and that enhanced production of peptidoleukotrienes causes reversible renal dysfunction. Increased leukotriene production within the kidney may therefore be important in the pathogenesis of lupus nephritis.

Authors
Spurney, RF; Ruiz, P; Pisetsky, DS; Coffman, TM
MLA Citation
Spurney, RF, Ruiz, P, Pisetsky, DS, and Coffman, TM. "Enhanced renal leukotriene production in murine lupus: role of lipoxygenase metabolites." Kidney Int 39.1 (January 1991): 95-102.
PMID
1848329
Source
pubmed
Published In
Kidney international
Volume
39
Issue
1
Publish Date
1991
Start Page
95
End Page
102

IgG binding of monoclonal anti-nuclear antibodies from MRL-lpr/lpr mice.

To assess the specificity of anti-nuclear antibodies with cross-reactive rheumatoid factor (RF) activity, monoclonal anti-DNA and anti-Sm antibodies from MRL-lpr/lpr mice were tested for binding to a variety of IgG antigens. These antibodies had all been previously identified as binding heterologous IgG. By ELISA, antibodies in this panel all bound BALB/c myeloma proteins representing the different IgG subclasses, indicating broad reactivity with murine IgG as well as heterologous IgG. The determinant recognized by these antibodies was further investigated using the Fab, F(ab')2 and Fc fragments of both human as well as rabbit origin. All antibodies bound well to fragments as well as intact IgG antigens. These antibodies were further analysed by Western blotting, demonstrating that most bound to both heavy and light chains of human origin. Together, these observations suggest that some anti-nuclear antibodies bind a conserved antigenic determinant present widely on immunoglobulin chains. This determinant may represent a common sequence important in immunoglobulin domain structure.

Authors
Pisetsky, DS; Darwin, BS; Reich, CF
MLA Citation
Pisetsky, DS, Darwin, BS, and Reich, CF. "IgG binding of monoclonal anti-nuclear antibodies from MRL-lpr/lpr mice." Immunology 71.4 (December 1990): 586-591.
PMID
2279742
Source
pubmed
Published In
Immunology
Volume
71
Issue
4
Publish Date
1990
Start Page
586
End Page
591

Heavy and light chain utilization in autoantibodies of elderly patients with systemic lupus erythematosus.

To determine whether age-related changes in immune function affect patterns of autoantibody production, we have examined the isotype and light chain utilization in autoantibodies of elderly patients with systemic lupus erythematosus (SLE). Enzyme-linked immunosorbent assays (ELISA) were used to determine the frequencies of IgG and IgM antibodies to single-stranded DNA (ssDNA), Sm, and the 70K protein component of RNP in the sera of 53 patients with SLE older than age 60. The IgG subclass distributions and kappa/lambda ratios for each of these autoantibodies were also determined and compared to measurements performed on the sera of 53 young adult patients with SLE. The frequencies of autoantibodies of each specificity, except IgM anti-ss DNA antibodies, were higher among the young adult patients, although the magnitudes of the responses were similar in both age groups. IgG anti-Sm antibodies were composed of both IgG1 and IgG2 subclasses, while IgG anti-70K RNP and IgG anti-ssDNA were predominantly of the IgG1 subclass. There were no differences in the IgG subclass distributions of any of the three autoantibodies between the elderly and young adult patient sera. The kappa/lambda ratios for each of the three autoantibodies were similar to that present in total serum immunoglobulins, and kappa/lambda ratios of autoantibodies, standardized to the kappa/lambda ratios of serum, were not different between elderly and young adult groups. Few patient sera of either age group (9 elderly, 7 young adult) demonstrated even midly skewed light chain ratios in their autoantibody responses. Thus, despite developing in an immunological environment that may have altered the clonality and isotype distribution of their responses, the autoantibodies produced by elderly patients with SLE were qualitatively similar to autoantibodies of younger patients.

Authors
Ward, MM; Pisetsky, DS
MLA Citation
Ward, MM, and Pisetsky, DS. "Heavy and light chain utilization in autoantibodies of elderly patients with systemic lupus erythematosus." Clin Immunol Immunopathol 57.2 (November 1990): 280-296.
PMID
2208808
Source
pubmed
Published In
Clinical Immunology and Immunopathology
Volume
57
Issue
2
Publish Date
1990
Start Page
280
End Page
296

Serum interleukin-2 receptor responses to immunization.

Serum interleukin-2 receptor (sIL-2R) levels have been used to assess immune activation in inflammatory and infectious illnesses, although the cellular origin of these receptors and the dynamics of their production are not well defined. To investigate the relationship between sIL-2R levels and the degree of immune activation in antigen-specific responses, sIL-2R were measured in healthy individuals after both primary and secondary immunization with keyhole limpet hemocyanin (KLH). Despite induction of strong antibody responses, KLH immunization did not result in consistent elevations of sIL-2R levels, with only one of six subjects developing a substantial (twofold) increase in sIL-2R levels. The absence of sIL-2R elevation after a discrete antigenic stimulus suggests that inflammatory illnesses in which elevated sIL-2R levels have been noted involve more extensive stimulation of immune cells, either in number or in degree, than that present after simple immunization in healthy individuals.

Authors
Ward, MM; Hall, RP; Pisetsky, DS
MLA Citation
Ward, MM, Hall, RP, and Pisetsky, DS. "Serum interleukin-2 receptor responses to immunization." Clin Immunol Immunopathol 57.1 (October 1990): 120-124.
PMID
2394031
Source
pubmed
Published In
Clinical Immunology and Immunopathology
Volume
57
Issue
1
Publish Date
1990
Start Page
120
End Page
124

Selective recognition of DNA antigenic determinants by murine monoclonal anti-DNA antibodies.

To assess the immune recognition of DNA in systemic lupus erythematosus, the antigenic specificity of monoclonal anti-DNA antibodies from autoimmune MRL-lpr/lpr mice was investigated Determinant specificity was assessed by ELISA in terms of binding to a panel of ssDNA antigens including calf thymus, human placenta, Escherichia coli, Clostridium perfringens, Micrococcus lysodeikticus, salmon testes, chicken blood and murine DNA. Among the monoclonal antibodies, a variety of binding patterns was observed, although for all antibodies tested murine DNA was among the most reactive antigens. Binding to other DNAs varied markedly, with some antibodies showing only low reactivity to certain antigens in the test panel. Similar results were obtained with sera of individual MRL-lpr/lpr mice. These results suggest that anti-DNA antibodies bind specific antigenic determinants variably expressed by DNAs of various species. Furthermore, the preferential binding to mouse DNA by some MRL-lpr/lpr antibodies may suggest a role of self-DNA in the in vivo selection of anti-DNA antibodies for expression.

Authors
Wu, DP; Gilkeson, GS; Armitage, J; Reich, CF; Pisetsky, DS
MLA Citation
Wu, DP, Gilkeson, GS, Armitage, J, Reich, CF, and Pisetsky, DS. "Selective recognition of DNA antigenic determinants by murine monoclonal anti-DNA antibodies." Clin Exp Immunol 82.1 (October 1990): 33-37.
PMID
1698581
Source
pubmed
Published In
Clinical & Experimental Immunology
Volume
82
Issue
1
Publish Date
1990
Start Page
33
End Page
37

Cellular requirements for anti-DNA production induced in mice by immunization with bacterial DNA.

To further define DNA immunization as a model for anti-DNA production, we investigated the cellular requirements for this response in mice immunized with single-stranded DNA from E. coli. The anti-DNA responses of genetically immune-deficient mice and congenic controls were measured by ELISA after immunization with E. coli DNA as complexes with methylated bovine serum albumin in complete Freund's adjuvant. T cell-deficient BALB/c-nu/nu mice failed to produce IgG anti-DNA by this protocol despite high backgrounds of IgM anti-DNA. In contrast, CBA/N mice expressing the xid defect displayed IgG anti-DNA responses comparable to those of CBA/J mice despite a reduced IgM response; the specificity of CBA/N and CBA/J anti-DNA antibodies was similar as determined by binding to synthetic DNA and RNA antigens. These results suggest that the anti-DNA response stimulated by DNA immunization is dependent on T cells but not the B cell population affected by xid. The intact IgG response of immunized xid mice differs from that of lupus mice bearing xid where this gene defect leads to significant reduction of spontaneous anti-DNA production.

Authors
Gilkeson, GS; Pritchard, AJ; Pisetsky, DS
MLA Citation
Gilkeson, GS, Pritchard, AJ, and Pisetsky, DS. "Cellular requirements for anti-DNA production induced in mice by immunization with bacterial DNA." Eur J Immunol 20.8 (August 1990): 1789-1794.
PMID
2209690
Source
pubmed
Published In
European Journal of Immunology
Volume
20
Issue
8
Publish Date
1990
Start Page
1789
End Page
1794
DOI
10.1002/eji.1830200825

The fine specificity of anti-La antibodies induced in mice by immunization with recombinant human La autoantigen.

Because of increasing evidence suggesting that anti-La autoantibodies are induced in humans by an Ag-specific mechanism, we investigated the antibody response of animals immunized with the human La Ag and studied its relationship to the anti-La response of autoimmune patients. Anti-La antibodies were raised in 6- to 8-wk-old male MRL(-)+/+, C57BL/6J, BALB/c, and A/J mice by immunizing with authentic human La protein obtained by recombinant expression in Escherichia coli. As we have shown previously for human autoantibodies, induced mouse anti-La antibodies reacted with recombinant fusion proteins containing nonoverlapping sequences from different portions of the La molecule. The epitope specificity of antibodies to the middle region of the La Ag was further evaluated using six synthetic La peptides predicted to be antigenic based on their hydrophilic properties. Although the induced mouse anti-La antibodies bound to five of the six synthetic La peptides, human anti-La autoantibodies failed to recognize any of the peptide homologs. These results suggest that mice respond to immunization with human La protein differently than humans who develop autoimmunity to this self Ag.

Authors
St Clair, EW; Kenan, D; Burch, JA; Keene, JD; Pisetsky, DS
MLA Citation
St Clair, EW, Kenan, D, Burch, JA, Keene, JD, and Pisetsky, DS. "The fine specificity of anti-La antibodies induced in mice by immunization with recombinant human La autoantigen." J Immunol 144.10 (May 15, 1990): 3868-3876.
PMID
1692063
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
144
Issue
10
Publish Date
1990
Start Page
3868
End Page
3876

Expression of IgM and IgG autoantibodies in pediatric and adult systemic lupus erythematosus.

To compare patterns of autoantibody responses in pediatric and adult patients with systemic lupus erythematosus (SLE). IgG and IgM antibodies to single-stranded DNA (ssDNA), Sm, and the 70-kDa protein component of the RNP antigen (70-kDa RNP) were measured in 29 pediatric and 36 adult patients by enzyme-linked immunosorbent assays. Antibodies of either isotype to ssDNA, Sm, and 70-kDa RNP were present in 64, 58, and 79% of pediatric patients, respectively, comparable to prevalences of these autoantibodies in the adult SLE patients. Pediatric SLE patients were more likely than adult patients to have IgM anti-Sm antibodies (41.4% vs 13.9%, P = 0.02) and tended to more commonly express IgM anti-70-kDa RNP and IgM anti-ssDNA antibodies. The prominence of IgM autoantibody responses among pediatric SLE patients was shown by multiple logistic regression analysis to be related to total IgM concentrations and not related to age or duration of disease. Sequential serum samples available from several pediatric patients revealed the maintenance of similar patterns of isotype responses over time in approximately one-half of patients. In those patients whose responses changed over time, the variations in isotype expression were consistent with maturation of antibody responses of each specificity. While these results demonstrate similarities in autoimmune reactivities between pediatric and adult SLE patients, the serologic study of pediatric patients may provide an opportunity to more readily investigate the evolution of autoantibody responses.

Authors
Ward, MM; Dawson, DV; Kredich, DW; Pisetsky, DS
MLA Citation
Ward, MM, Dawson, DV, Kredich, DW, and Pisetsky, DS. "Expression of IgM and IgG autoantibodies in pediatric and adult systemic lupus erythematosus." Clin Immunol Immunopathol 55.2 (May 1990): 273-284.
PMID
2323106
Source
pubmed
Published In
Clinical Immunology and Immunopathology
Volume
55
Issue
2
Publish Date
1990
Start Page
273
End Page
284

Correlation between erythrocyte CR1 reduction and other blood proteinase markers in patients with malignant and inflammatory disorders.

Erythrocyte CR1, a C3b/C4b-binding complement-regulatory protein, is sensitive to proteolysis in vitro. To test the hypothesis that in vivo erythrocyte CR1 reduction results from intravascular proteinase activities, we used enzyme-linked immunosorbent assays to measure gamma-crosslinked fibrin degradation products (D-dimers) as indicators of coagulation/fibrinolytic activity, and complexes of neutrophil elastase with alpha 1 proteinase inhibitor (E/A) as indicators of neutrophil enzyme release in malignant and inflammatory disorders. Erythrocyte CR1, measured by monoclonal anti-CR1 antibody binding, was inversely related to disease activity and blood proteinase markers. Levels of erythrocyte CR1 were significantly lower for patients with active versus remittent squamous and small cell lung cancers, Hodgkin's and diffuse large cell lymphomas, and acute myelogenous leukemias. In patients with active thoracic cancers, elevated D-dimer levels correlated with reduction of CR1. In patients with rheumatoid arthritis, CR1 reduction was correlated with elevated levels of elastase complexes. Our findings substantiate the relationship of acquired CR1 reduction to the activity of certain diseases and provide circumstantial support for the hypothesis that erythrocyte CR1 is lost to proteolysis in vivo. Although heritable differences in CR1 expression reduce the interpretability of single measurements of erythrocyte CR1 levels, disease-associated CR1 reduction may be a useful indicator of disorders with chronically increased blood proteinase activity.

Authors
Currie, MS; Vala, M; Pisetsky, DS; Greenberg, CS; Crawford, J; Cohen, HJ
MLA Citation
Currie, MS, Vala, M, Pisetsky, DS, Greenberg, CS, Crawford, J, and Cohen, HJ. "Correlation between erythrocyte CR1 reduction and other blood proteinase markers in patients with malignant and inflammatory disorders." Blood 75.8 (April 15, 1990): 1699-1704.
PMID
2158365
Source
pubmed
Published In
Blood
Volume
75
Issue
8
Publish Date
1990
Start Page
1699
End Page
1704

In vitro autoantibody production by normal adult and cord blood B cells.

To investigate the repertoire of autoantibodies in humans, anti-DNA and rheumatoid factor (RF) production in vitro was assessed in cultures of adult peripheral blood B cells and neonatal umbilical venous blood B cells. B cells were stimulated under various culture conditions, using an immobilized monoclonal anti-CD3 antibody and adult T cells or Staphylococcus aureus (SA) in the presence or absence of adult T cells or factors derived from mitogen-stimulated adult T cells as polyclonal B cell activators. Total IgM, as well as IgM anti-DNA and RF, were assessed by ELISA. Total IgM production was induced from adult and neonatal B cells with SA plus T cell factors, as well as anti-CD3-stimulated T cells. RF was induced from adult and cord blood B cells by either mode of stimulation, whereas significant anti-DNA production was observed only when B cells were stimulated with anti-CD3-activated T cells. These results confirm the presence of B cell precursors for autoantibodies in the preimmune as well as normal adult repertoire, and indicate that the production of anti-DNA and RF appears to be regulated independently.

Authors
Pisetsky, DS; Jelinek, DF; McAnally, LM; Reich, CF; Lipsky, PE
MLA Citation
Pisetsky, DS, Jelinek, DF, McAnally, LM, Reich, CF, and Lipsky, PE. "In vitro autoantibody production by normal adult and cord blood B cells." J Clin Invest 85.3 (March 1990): 899-903.
PMID
2138166
Source
pubmed
Published In
Journal of Clinical Investigation
Volume
85
Issue
3
Publish Date
1990
Start Page
899
End Page
903
DOI
10.1172/JCI114517

A role for immunogenic DNA in the pathogenesis of systemic lupus erythematosus.

Authors
Pisetsky, DS; Grudier, JP; Gilkeson, GS
MLA Citation
Pisetsky, DS, Grudier, JP, and Gilkeson, GS. "A role for immunogenic DNA in the pathogenesis of systemic lupus erythematosus." Arthritis Rheum 33.2 (February 1990): 153-159.
PMID
2407246
Source
pubmed
Published In
Arthritis and Rheumatism
Volume
33
Issue
2
Publish Date
1990
Start Page
153
End Page
159

Expression of autoantibodies to recombinant (U1) RNP-associated 70K antigen in systemic lupus erythematosus.

To determine the specificity of antibodies to the (U1) ribonucleoprotein antigen in systemic lupus erythematosus (SLE), patient sera were tested for binding to a recombinant human 70K antigen. By solid-phase immunoassay, we detected anti-70K reactivity in sera from 31 of 96 patients with systemic lupus erythematosus (SLE), demonstrating that anti-70K antibodies may occur in patients with SLE as well as other clinical diagnoses. In sequential sera from 2 of these patients, we found that anti-70K binding varied dramatically over the course of disease. The changes in anti-70K antibody levels did not correlate with clinical events nor evolving antibody reactivity with the Sm-specific antigens.

Authors
St Clair, EW; Query, CC; Bentley, R; Keene, JD; Polisson, RP; Allen, NB; Caldwell, DS; Rice, JR; Cox, C; Pisetsky, DS
MLA Citation
St Clair, EW, Query, CC, Bentley, R, Keene, JD, Polisson, RP, Allen, NB, Caldwell, DS, Rice, JR, Cox, C, and Pisetsky, DS. "Expression of autoantibodies to recombinant (U1) RNP-associated 70K antigen in systemic lupus erythematosus." Clin Immunol Immunopathol 54.2 (February 1990): 266-280.
PMID
2104788
Source
pubmed
Published In
Clinical Immunology and Immunopathology
Volume
54
Issue
2
Publish Date
1990
Start Page
266
End Page
280

Temporal correlation of antibody responses to different epitopes of the human La autoantigen.

To investigate the temporal relationship of antibody responses to different La epitopes, sequential sera from nine patients with systemic lupus erythematosus and Sjogren's syndrome were tested by enzyme-linked immunosorbent assay for antibody binding to a series of recombinant fusion proteins containing different regions of the La molecule. The results of this analysis indicate that antibody responses to four different La fragments vary in parallel over time. This finding is supported by a statistical analysis indicating that the changes in antibody levels between the six pairs of responses were highly correlated (P less than 0.001). Furthermore, we show by immunoaffinity purification that antibodies to the three nonoverlapping La protein fragments do not cross-react with other fragments and, hence, represent independent populations. These results suggest that anti-La antibodies are coordinately produced to different epitopes on the La molecule, possibly reflecting an antigen-driven mechanism.

Authors
St Clair, EW; Burch, JA; Ward, MM; Keene, JD; Pisetsky, DS
MLA Citation
St Clair, EW, Burch, JA, Ward, MM, Keene, JD, and Pisetsky, DS. "Temporal correlation of antibody responses to different epitopes of the human La autoantigen." J Clin Invest 85.2 (February 1990): 515-521.
PMID
1688887
Source
pubmed
Published In
Journal of Clinical Investigation
Volume
85
Issue
2
Publish Date
1990
Start Page
515
End Page
521
DOI
10.1172/JCI114467

Regulation of the anti-Sm autoantibody response in systemic lupus erythematosus mice by monoclonal anti-Sm antibodies.

The administration of certain monoclonal anti-Sm antibodies (2G7, 7.13) induced most MRL/lpr mice to become anti-Sm positive by 5 mo of age, although other anti-Sm monoclonals (Y2, Y12) suppressed the spontaneous response. Positive anti-Sm antibody enhancement occurred efficiently only in MRL/lpr mice and not in other systemic lupus erythematosus mice that have little spontaneous anti-Sm production. The enhancement by anti-Sm antibodies was specific for the anti-Sm response. The mechanism of the passive antibody enhancement was apparently not isotype- or idiotype-related. The fine specificity of the anti-Sm monoclonal antibody may be essential to its enhancing or suppressing effects, since both enhancing monoclonals recognized only the D Sm polypeptide, whereas both suppressing monoclonals saw the D and the B polypeptides. Furthermore, analysis of serial bleeds from unmanipulated MRL mice that developed anti-Sm positivity showed that the D specificity almost always appeared first. We hypothesize, therefore, that those animals in which an anti-Sm response is initiated by D-specific B-cell clones can become serologically positive with the aid of a positive feedback loop. In contrast, animals in which the initial specificity is for both B and D peptides would be prevented from developing a full anti-Sm response.

Authors
Eisenberg, RA; Pisetsky, DS; Craven, SY; Grudier, JP; O'Donnell, MA; Cohen, PL
MLA Citation
Eisenberg, RA, Pisetsky, DS, Craven, SY, Grudier, JP, O'Donnell, MA, and Cohen, PL. "Regulation of the anti-Sm autoantibody response in systemic lupus erythematosus mice by monoclonal anti-Sm antibodies." J Clin Invest 85.1 (January 1990): 86-92.
PMID
2295704
Source
pubmed
Published In
Journal of Clinical Investigation
Volume
85
Issue
1
Publish Date
1990
Start Page
86
End Page
92
DOI
10.1172/JCI114437

Anti-DNA antibodies from autoimmune mice arise by clonal expansion and somatic mutation

Authors
Shlomchik, M; Mascelli, M; Shan, H; Radic, MZ; Pisetsky, D; Marshak-Rothstein, A; Weigert, M
MLA Citation
Shlomchik, M, Mascelli, M, Shan, H, Radic, MZ, Pisetsky, D, Marshak-Rothstein, A, and Weigert, M. "Anti-DNA antibodies from autoimmune mice arise by clonal expansion and somatic mutation." Journal of Experimental Medicine 171.1 (1990): 265-297.
PMID
2104919
Source
scival
Published In
The Journal of Experimental Medicine
Volume
171
Issue
1
Publish Date
1990
Start Page
265
End Page
297

Polyspecific binding of Escherichia coli beta-galactosidase by murine antibodies to DNA.

To characterize further polyspecific interactions of antibodies to DNA, the binding of sera from autoimmune MRL-lpr/lpr mice to Escherichia coli beta-galactosidase (beta-gal) was analyzed. This protein was selected for study because of preliminary observations that sera from autoimmune mice bound unexpectedly to cloned fusion protein constructions containing beta-gal. Using ELISA assays, sera from MRL-lpr/lpr mice demonstrated high levels of antibodies to both DNA and beta-gal, in titers significantly greater than those of BALB/c controls. Affinity chromatography using beta-gal-Sepharose demonstrated that antibodies enriched for anti-beta-gal activity bound both DNA as well as beta-gal, indicating the presence of a population of cross-reactive anti-DNA antibodies. Furthermore, anti-DNA mAb of MRL-lpr/lpr strain origin also bound beta-gal by ELISA, although these levels were lower than those to DNA. Together, these results extend the range of polyspecific binding of murine anti-DNA antibodies to bacterial proteins. They further suggest caution in the interpretation of immunoassays using fusion protein constructions containing beta-gal, especially with sera from autoimmune mice.

Authors
Pisetsky, DS; Grudier, JP
MLA Citation
Pisetsky, DS, and Grudier, JP. "Polyspecific binding of Escherichia coli beta-galactosidase by murine antibodies to DNA." J Immunol 143.11 (December 1, 1989): 3609-3613.
PMID
2511245
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
143
Issue
11
Publish Date
1989
Start Page
3609
End Page
3613

Genetic control of inflammatory arthritis in congenic lpr mice.

To determine the genetic requirements for the development of inflammatory arthritis in MRL-lpr/lpr mice, clinical, serologic, and pathologic features of lpr/lpr and +/+ mice of MRL, B6, C3H, and AKR strains were studied. Arthritis was evaluated by histopathologic examination of the knee joint, while sera were tested for the presence of rheumatoid factor (RF) and anti-DNA activity by ELISA. Of the strains tested to age 7 months, only the MRL-lpr/lpr mice developed histologic evidence of arthritis. All lpr mice, however, produced both IgM RF and IgG RF, although amounts varied among strains. These results indicate that the lpr gene as well as another gene(s) in the MRL background are necessary for the development of inflammatory arthritis and that this lesion may be independent of RF production.

Authors
Gilkeson, GS; Ruiz, P; Grudier, JP; Kurlander, RJ; Pisetsky, DS
MLA Citation
Gilkeson, GS, Ruiz, P, Grudier, JP, Kurlander, RJ, and Pisetsky, DS. "Genetic control of inflammatory arthritis in congenic lpr mice." Clin Immunol Immunopathol 53.3 (December 1989): 460-474.
PMID
2805451
Source
pubmed
Published In
Clinical Immunology and Immunopathology
Volume
53
Issue
3
Publish Date
1989
Start Page
460
End Page
474

Patterns of autoantibody expression in pediatric and adult systemic lupus erythematosus.

To characterize patterns of autoantibody expression in pediatric and adult systemic lupus erythematosus (SLE), IgG and IgM antibodies to cardiolipin (aCL) and single-stranded DNA (anti-DNA) were measured in sera from 32 pediatric and 36 adult patients. Antibody levels were assessed by isotype-specific ELISA and compared with 15 pediatric disease controls and 36 healthy adult controls. These determinations revealed significant differences in the pattern of IgG and IgM autoantibody expression in adult and pediatric patients. IgM aCL were more common in the pediatric than adult SLE population with 11 of 32 pediatric patients being positive compared to only 4 of 36 in the adult group (p less than 0.05). Conversely, enhanced IgG autoantibody production was observed in the adult group as 27 of 36 adult patients were positive for IgG anti-DNA compared to only 16 of 32 children (p less than 0.05). Analysis of sequential sera showed differences in the temporal expression of IgG and IgM autoantibodies as well as variability between anti-DNA and aCL responses over time. Our data suggest differences in the isotype profile of autoantibody responses in pediatric and adult SLE and provide further evidence for heterogeneity in the mechanisms of autoantibody production.

Authors
Shergy, WJ; Kredich, DW; Pisetsky, DS
MLA Citation
Shergy, WJ, Kredich, DW, and Pisetsky, DS. "Patterns of autoantibody expression in pediatric and adult systemic lupus erythematosus." J Rheumatol 16.10 (October 1989): 1329-1334.
PMID
2810258
Source
pubmed
Published In
The Journal of rheumatology
Volume
16
Issue
10
Publish Date
1989
Start Page
1329
End Page
1334

The genetics of autoantibody production in MRL/lpr lupus mice.

We have investigated the influence of background genes in the MRL strain, as compared to C57BL/6, on the induction of autoimmunity in homozygous lpr/lpr mice. We have concentrated on two autoantibodies, anti-Sm and anti-chromatin. The propensity to make anti-Sm is controlled by dominant genes from the MRL background. However, the prevalence of this response is under the control of additional recessive genes. The anti-chromatin response is found in both MRL/Mp-lpr/lpr and in C57BL/6-lpr/lpr mice, but it appears earlier and in higher titers in the former strain. This high responder effect is controlled by dominant genes. In F1 mice between these two strains, both anti-Sm and anti-chromatin antibodies preferentially use the b Igh allotype from the low responder B6/lpr parent. In addition, in the progeny of backcross of the F1 to the MRL/lpr strain, the production of both anti-Sm and anti-chromatin is linked to the b allotype. These results demonstrate the contribution of dominant genes from the MRL background on the induction of severe autoimmunity. They also suggest that the B6 background expresses an Igh allotype particularly amenable to autoantibody production, in spite of the relatively mild SLE-like syndrome in this strain.

Authors
Eisenberg, RA; Craven, SY; Fisher, CL; Morris, SC; Rapoport, R; Pisetsky, DS; Cohen, PL
MLA Citation
Eisenberg, RA, Craven, SY, Fisher, CL, Morris, SC, Rapoport, R, Pisetsky, DS, and Cohen, PL. "The genetics of autoantibody production in MRL/lpr lupus mice." Clin Exp Rheumatol 7 Suppl 3 (September 1989): S35-S40.
PMID
2514059
Source
pubmed
Published In
Clinical and experimental rheumatology
Volume
7 Suppl 3
Publish Date
1989
Start Page
S35
End Page
S40

Epitope specificity of anti-La antibodies from patients with Sjögren's syndrome.

To investigate patterns of autoreactivity in Sjögren's syndrome, the epitope specificity of anti-La antibodies was determined using recombinant antigens bearing sequences of the amino, middle, and carboxyl portions of the La molecule. Sera from patients with primary as well as secondary Sjögren's syndrome reacted with all three fragments, although the magnitude of the responses varied markedly among individuals. Furthermore, the proportion of antibody binding directed to the different La epitopes showed considerable individual variations, but these patterns were not correlated with specific clinical manifestations. These results suggest that quantitative and qualitative aspects of anti-La responses in Sjögren's syndrome are determined by factors distinct from those determining the clinical expression of disease.

Authors
St Clair, EW; Talal, N; Moutsopoulos, HM; Ballester, A; Zerva, L; Keene, JD; Pisetsky, DS
MLA Citation
St Clair, EW, Talal, N, Moutsopoulos, HM, Ballester, A, Zerva, L, Keene, JD, and Pisetsky, DS. "Epitope specificity of anti-La antibodies from patients with Sjögren's syndrome." J Autoimmun 2.4 (August 1989): 335-344.
PMID
2477000
Source
pubmed
Published In
Journal of Autoimmunity
Volume
2
Issue
4
Publish Date
1989
Start Page
335
End Page
344

The antibody response of normal mice to immunization with single-stranded DNA of various species origin.

To further assess the mechanism for the induction of anti-DNA antibodies, the response of BALB/c mice to immunization with single-stranded DNA of various species origin was determined. Anti-DNA levels of mice immunized with Escherichia coli DNA as complexes with methylated BSA in adjuvant were significantly greater by ELISA than those from mice immunized similarly with calf thymus DNA. Furthermore, comparison of the responses of mice immunized with complexes of DNA from calf thymus, chicken blood, Clostridium perfringens, E. coli, human placenta, or salmon testes indicated that the bacterial DNAs induced the highest antibody levels. The antibody response to E. coli DNA was shown by inhibition ELISA to have two populations, one binding unique determinants in E. coli DNA and the other cross-reactive with determinants expressed on all DNAs tested. These results indicate that DNA molecules, when complexed to a protein carrier, differ in their immunogenic potential, likely because of the presence of unique sequences or structures rarely presented by mammalian host DNA.

Authors
Gilkeson, GS; Grudier, JP; Pisetsky, DS
MLA Citation
Gilkeson, GS, Grudier, JP, and Pisetsky, DS. "The antibody response of normal mice to immunization with single-stranded DNA of various species origin." Clin Immunol Immunopathol 51.3 (June 1989): 362-371.
PMID
2785884
Source
pubmed
Published In
Clinical Immunology and Immunopathology
Volume
51
Issue
3
Publish Date
1989
Start Page
362
End Page
371

Antidouble stranded DNA antibody assays in systemic lupus erythematosus: correlations of longitudinal antibody measurements.

To determine whether different assays of antidouble stranded DNA (anti-dsDNA) antibodies provide comparable information in quantitative antibody assessment over time, longitudinal correlations between 3 anti-dsDNA antibody methods were derived. Determinations of anti-dsDNA antibody levels on serial samples from 9 patients with systemic lupus erythematosus (SLE) were performed by filter binding radioimmunoassay, enzyme linked immunosorbent assay, and Crithidia indirect immunofluorescence. Substantial pairwise correlations among assay methods were found (r = 0.544 to 0.804; p less than 0.001). In addition, anti-dsDNA antibody levels as measured by each assay were inversely correlated with levels of the 3rd component of complement. Our results indicate that changes in antibody levels as determined by these 3 methods closely parallel each other over time, and suggest that the array of anti-dsDNA antibodies detected in patient sera remains relatively constant over time.

Authors
Ward, MM; Pisetsky, DS; Christenson, VD
MLA Citation
Ward, MM, Pisetsky, DS, and Christenson, VD. "Antidouble stranded DNA antibody assays in systemic lupus erythematosus: correlations of longitudinal antibody measurements." J Rheumatol 16.5 (May 1989): 609-613.
PMID
2666655
Source
pubmed
Published In
The Journal of rheumatology
Volume
16
Issue
5
Publish Date
1989
Start Page
609
End Page
613

Induction of anti-double stranded DNA antibodies in normal mice by immunization with bacterial DNA.

Because of recent observations suggesting that bacterial DNA is immunogenic, the induction in normal mice of antibodies to Escherichia coli (EC) dsDNA was investigated. BALB/c and C57BL/6 mice were immunized with dsEC or ds calf thymus (CT) DNA complexed to methylated BSA in adjuvant; antibody responses were measured by ELISA. In both strains, dsEC DNA immunization induced a much higher anti-dsDNA response to dsEC DNA than did dsCT DNA immunization. Neither immunized group showed an appreciable antibody response when tested on dsCT DNA. Anti-dsDNA antibodies were also demonstrated by ELISA using synthetic DNA duplexes as well as a filter binding assay using 3H-labeled dsEC DNA as Ag. These results suggest that bacterial dsDNA is immunogenic and that at least some anti-dsDNA specificities can arise by immunization.

Authors
Gilkeson, GS; Grudier, JP; Karounos, DG; Pisetsky, DS
MLA Citation
Gilkeson, GS, Grudier, JP, Karounos, DG, and Pisetsky, DS. "Induction of anti-double stranded DNA antibodies in normal mice by immunization with bacterial DNA." J Immunol 142.5 (March 1, 1989): 1482-1486.
PMID
2645362
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
142
Issue
5
Publish Date
1989
Start Page
1482
End Page
1486

Analysis of autoantibody binding to different regions of the human La antigen expressed in recombinant fusion proteins.

To determine the specificity of autoantibodies for various antigenic sites on a self-protein molecule, sera from 19 patients with anti-La antibodies were tested for their reactivity with molecularly cloned La protein fragments. By quantitative ELISA, anti-La sera from patients with various connective tissue diseases were shown to react with La fusion proteins containing different regions of the La molecule. Two recombinant La fragments containing the carboxyl three-fourths and the middle one-third of the La sequence, respectively, bound higher levels of anti-La antibodies than the two fragments representing the amino and carboxyl terminals. Purified bovine La protein effectively competed for the binding of human autoantibodies to three of the four recombinant La fusion proteins, suggesting similarity in antigenic presentation between the La epitopes in these fusion proteins and the native La molecule. Immunoadsorption experiments showed that most anti-bovine La protein antibodies were removed from a human serum by affinity chromatography by using the fusion protein containing the carboxyl three-fourths of the La sequence, thus supporting the results obtained by quantitative solid phase ELISA. These studies demonstrate that anti-La autoantibodies recognize three La fragments representing separate nonoverlapping regions of the La sequence and are compatible with a mechanism of autoantibody production based on an immune response to the entire self-protein molecule.

Authors
St Clair, EW; Pisetsky, DS; Reich, CF; Keene, JD
MLA Citation
St Clair, EW, Pisetsky, DS, Reich, CF, and Keene, JD. "Analysis of autoantibody binding to different regions of the human La antigen expressed in recombinant fusion proteins." J Immunol 141.12 (December 15, 1988): 4173-4180.
PMID
2461985
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
141
Issue
12
Publish Date
1988
Start Page
4173
End Page
4180

Methotrexate-associated hepatotoxicity: retrospective analysis of 210 patients with rheumatoid arthritis.

PURPOSE: Beginning in the 1980s, methotrexate has been used successfully to treat rheumatoid arthritis. The magnitude and severity of short- and long-term methotrexate toxicity, however, have not been adequately investigated. Our study was performed to determine the prevalence of hepatotoxicity in patients with rheumatoid arthritis receiving long-term methotrexate therapy. PATIENTS AND METHODS: We conducted a retrospective, computer-assisted review of all Duke University Medical Center patients undergoing liver biopsy for methotrexate monitoring from January 1979 to January 1988. A total of 538 biopsies were performed in 399 patients, 259 of whom had inflammatory arthritis (210 with rheumatoid arthritis, 47 with psoriatic arthritis, and two with seronegative spondyloarthropathy). RESULTS: No evidence of cirrhosis was defined in the cohort with rheumatoid arthritis; however, six patients with rheumatoid arthritis had histologic changes of fibrotic liver disease (prevalence of 2.9 percent in the group with rheumatoid arthritis) while taking methotrexate. Five of the six patients were obese and three had glucose intolerance or overt diabetes mellitus, and one person admitted to alcohol usage. Only one patient with fibrotic liver disease had elevated liver function test results, and no person showed a declining serum albumin level at the time of biopsy. Sixty-one patients with rheumatoid arthritis underwent multiple samplings (44 with two, 13 with three, and four with four biopsies). Fourteen of these patients showed progressive hepatic disease, whereas four patients improved. CONCLUSION: Although the prevalence of methotrexate hepatotoxicity in this large cohort of patients with rheumatoid arthritis was low, a small but definite risk of hepatic fibrosis, not predictable by laboratory screening, still exists.

Authors
Shergy, WJ; Polisson, RP; Caldwell, DS; Rice, JR; Pisetsky, DS; Allen, NB
MLA Citation
Shergy, WJ, Polisson, RP, Caldwell, DS, Rice, JR, Pisetsky, DS, and Allen, NB. "Methotrexate-associated hepatotoxicity: retrospective analysis of 210 patients with rheumatoid arthritis." Am J Med 85.6 (December 1988): 771-774.
PMID
3195601
Source
pubmed
Published In
The American Journal of Medicine
Volume
85
Issue
6
Publish Date
1988
Start Page
771
End Page
774

Immunization with the Sm nuclear antigen induces anti-Sm antibodies in normal and MRL mice.

The spontaneous occurrence of antibodies against the Sm nuclear antigen is a highly specific marker for the diagnosis of SLE. We have previously shown that anti-Sm can be elicited by immunization of SLE-prone mice with purified Sm antigen. In the present study, this autoantibody was induced in normal mice by a similar immunization protocol. Anti-Sm produced by normal strains was predominantly IgG1, which is similar to the isotype distribution in Sm-immunized MRL mice, but unlike the IgG2a-dominated response seen for spontaneous anti-Sm. Anti-Sm raised by immunization in most strains recognized epitopes not seen by spontaneous human and murine SLE anti-Sm; of the eleven normal strains tested, only C3H and AKR, strains from which MRL was partially derived, responded to these determinants. Further, immunoblot analysis of anti-Sm generated by immunization of MRL and normal mice revealed that the same proteins recognized by spontaneous human and murine anti-Sm were also seen by these sera. This study shows that an autoantibody highly characteristic of SLE can be produced in normal and MRL mice after appropriate immunization, and that the fine specificity of such experimentally induced antibody can be similar to that of spontaneous anti-Sm autoantibodies. The results imply a role for autoimmunization with Sm in the production of anti-Sm.

Authors
Shores, EW; Pisetsky, DS; Grudier, J; Eisenberg, RA; Cohen, PL
MLA Citation
Shores, EW, Pisetsky, DS, Grudier, J, Eisenberg, RA, and Cohen, PL. "Immunization with the Sm nuclear antigen induces anti-Sm antibodies in normal and MRL mice." Immunology 65.3 (November 1988): 473-478.
PMID
3264813
Source
pubmed
Published In
Immunology
Volume
65
Issue
3
Publish Date
1988
Start Page
473
End Page
478

Mike's treatment.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Mike's treatment." Am J Med 85.4 (October 1988): 543-.
PMID
3177403
Source
pubmed
Published In
The American Journal of Medicine
Volume
85
Issue
4
Publish Date
1988
Start Page
543

The relationship of anticardiolipin antibodies to disease manifestations in pediatric systemic lupus erythematosus.

To determine the prevalence of anticardiolipin antibodies (aCL) in pediatric systemic lupus erythematosus (SLE) and their possible association with clinical manifestations, aCL were measured in sera of 32 patients with the onset of SLE before age 16. IgM and IgG aCL were determined by ELISA and values compared to those of 12 patients with juvenile rheumatoid arthritis (JRA), 15 age matched asthmatics, and 32 adult controls. aCL were demonstrated in sera of 16 of 32 (50%) children with SLE, 5 of 12 (42%) patients with JRA, 1 of 15 (7%) asthmatics, and in none of the 32 adult controls. Serial samples on 11 patients with SLE showed fluctuations in aCL levels that often corresponded to disease activity; the highest levels occurred in patients during periods of seizure activity and other neurologic events. The antibodies were not crossreactive anti-DNA antibodies as shown by the failure of DNA to inhibit binding to cardiolipin. These data suggest that the prevalence of aCL is similar in pediatric and adult SLE and that aCL levels may vary with disease activity, especially neurologic disease.

Authors
Shergy, WJ; Kredich, DW; Pisetsky, DS
MLA Citation
Shergy, WJ, Kredich, DW, and Pisetsky, DS. "The relationship of anticardiolipin antibodies to disease manifestations in pediatric systemic lupus erythematosus." J Rheumatol 15.9 (September 1988): 1389-1394.
PMID
3264338
Source
pubmed
Published In
The Journal of rheumatology
Volume
15
Issue
9
Publish Date
1988
Start Page
1389
End Page
1394

Quantitative immunoassay of anti-La antibodies using purified recombinant La antigen.

A purified recombinant La fusion protein was tested in an enzyme-linked immunosorbent assay to quantitate anti-La responses. This protein contained the immunodominant region of the La molecule fused to beta-galactosidase. In solid-phase assays, recombinant La protein was solubilized in urea and bound to polystyrene wells without loss of immunoreactivity. The recombinant-based enzyme-linked immunosorbent assay proved to be a sensitive method for the detection of anti-La binding, and it accurately distinguished anti-La precipitin positive sera from normal sera.

Authors
St Clair, EW; Pisetsky, DS; Reich, CF; Chambers, JC; Keene, JD
MLA Citation
St Clair, EW, Pisetsky, DS, Reich, CF, Chambers, JC, and Keene, JD. "Quantitative immunoassay of anti-La antibodies using purified recombinant La antigen." Arthritis Rheum 31.4 (April 1988): 506-514.
PMID
3128988
Source
pubmed
Published In
Arthritis and Rheumatism
Volume
31
Issue
4
Publish Date
1988
Start Page
506
End Page
514

Spontaneous expression of antibodies to DNA of various species origin in sera of normal subjects and patients with systemic lupus erythematosus.

To investigate mechanisms for the induction of anti-DNA antibodies in systemic lupus erythematosus (SLE), the specificity of anti-DNA antibodies was determined in sera from SLE patients and normal control subjects. As a marker of these responses, the reactivity to single-stranded DNA of various mammalian and bacterial species origin was tested by enzyme-linked immunosorbent assay. Patients with SLE demonstrated serum antibodies to all six types of DNA tested, whereas normal control subjects showed appreciable antibody responses only to DNA obtained from Micrococcus lysodeikticus (MC) and Staphylococcus epidermidus (SE). Anti-DNA antibodies in normal sera appeared to recognize unique sites on the DNA because MC DNA failed to inhibit antibody binding to SE DNA, and vice versa; in contrast, SLE antibody binding to MC DNA could be inhibited by SE as well as other DNA, suggesting recognition of a more widely shared epitope. The expression in normal sera of antibodies specific for certain bacterial DNA is consistent with their induction by structural determinants on these DNA molecules that are immunogenic. DNA may therefore represent another bacterial macromolecule capable of inducing cross-reactive antibodies in human autoimmune disease.

Authors
Karounos, DG; Grudier, JP; Pisetsky, DS
MLA Citation
Karounos, DG, Grudier, JP, and Pisetsky, DS. "Spontaneous expression of antibodies to DNA of various species origin in sera of normal subjects and patients with systemic lupus erythematosus." J Immunol 140.2 (January 15, 1988): 451-455.
PMID
3257238
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
140
Issue
2
Publish Date
1988
Start Page
451
End Page
455

Mechanisms of anti-DNA production in human and murine SLE.

Antibodies to DNA occur prominently in systemic lupus erythematosus and play a central role in pathogenesis. Recent studies on the production of anti-DNA antibodies in both mouse and man suggest that this response can result from a variety of immunoregulatory disturbances of B and/or T cells. The immunogenic role of DNA as opposed to other self or foreign antigens remains unclear, however. Future studies will delineate the structure of anti-DNA antibodies and thereby determine the role of antigen drive in this response.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Mechanisms of anti-DNA production in human and murine SLE." In Vivo 2.1 (January 1988): 31-34. (Review)
PMID
2979813
Source
pubmed
Published In
In vivo (Athens, Greece)
Volume
2
Issue
1
Publish Date
1988
Start Page
31
End Page
34

Inhibition of in vitro NZB antibody responses by cyclosporine.

To characterize functional abnormalities of B cells in murine autoimmunity, the effect of cyclosporine (CsA) on antibody responses of NZB and control BALB/c spleen cells was investigated in vitro. Under conditions of high cell density, both NZB and BALB/c spleen cells spontaneously produced IgM and IgM anti-DNA, although NZB responses were greater in magnitude and demonstrated at lower cell densities. In cultures at 5 x 10(6) cells/ml, CsA preferentially inhibited anti-DNA production by cells of both strains. However, to achieve similar levels of inhibition of both responses, significantly higher concentrations of CsA were required for cultures of NZB compared to BALB/c cells. These results support previous experiments indicating enhanced sensitivity to CsA as a pharmacological marker of autoantibody producing B cells. They further suggest that B cell activation in certain forms of autoimmunity may be associated with decreased sensitivity to immunosuppressive agents.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Inhibition of in vitro NZB antibody responses by cyclosporine." Clin Exp Immunol 71.1 (January 1988): 155-158.
PMID
3258202
Source
pubmed
Published In
Clinical & Experimental Immunology
Volume
71
Issue
1
Publish Date
1988
Start Page
155
End Page
158

Mike's treatment

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Mike's treatment." The American Journal of Medicine 85.C (1988): 543--.
Source
scival
Published In
The American Journal of Medicine
Volume
85
Issue
C
Publish Date
1988
Start Page
543-

Mechanisms of antinuclear antibody production in the rheumatic diseases.

Recent investigations on the mechanisms of ANA production in the rheumatic diseases suggest that these responses frequently emerge in the setting of nonspecific immunoregulatory disturbances. However, the expansion and maturation of these responses to generate pathogenic specificities may require the coexistence of clonal or antigen-specific abnormalities. At the genetic level, ANAs appear to be of diverse origin, including some with representation in the normal B cell repertoire. Since some ANAs are disease-specific, elucidation of the basis of their production should provide insights into the unique pathogenetic features of different rheumatic diseases.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Mechanisms of antinuclear antibody production in the rheumatic diseases." Rheum Dis Clin North Am 13.3 (December 1987): 569-581. (Review)
PMID
3324206
Source
pubmed
Published In
Rheumatic Disease Clinics of North America
Volume
13
Issue
3
Publish Date
1987
Start Page
569
End Page
581

Structure and function of anti-DNA autoantibodies derived from a single autoimmune mouse.

Four monoclonal anti-DNA antibodies derived from a single autoimmune MRL/lpr mouse were studied. Three of these antibodies showed similarities in DNA binding; the fourth had a much higher specific activity for single-stranded DNA and, in addition, was unique in binding double-stranded DNA and cardiolipin. Complete nucleotide sequences of heavy- and light-chain variable regions demonstrated that all four antibodies are clonally related. The sequences also showed numerous somatic mutations, the distribution of which suggests that positive selection by antigen operated on these clonally related autoantibodies.

Authors
Shlomchik, MJ; Aucoin, AH; Pisetsky, DS; Weigert, MG
MLA Citation
Shlomchik, MJ, Aucoin, AH, Pisetsky, DS, and Weigert, MG. "Structure and function of anti-DNA autoantibodies derived from a single autoimmune mouse." Proc Natl Acad Sci U S A 84.24 (December 1987): 9150-9154.
PMID
3480535
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
84
Issue
24
Publish Date
1987
Start Page
9150
End Page
9154

Polyspecific reactivity of a murine monoclonal antibody that binds to nuclear matrix-associated, chromatin-bound autoantigens.

To investigate polyspecific autoantibody interactions, we have characterized the binding of a cloned murine monoclonal IgM antibody termed (RTE-23) of strain BALB/c origin. By indirect immunofluorescence this antibody displayed a nuclear speckled and peripheral pattern in interphase cells from human and rodent cell lines. In contrast, in mitotic cells, antibody RTE-23 bound to the periphery of individual chromosomes. Immunoblot analysis of soluble and insoluble nuclear proteins from purified rat fibroblast nuclei showed that antibody RTE-23 bound to molecules of 28, 29, and 33 kDa. Furthermore, antibody RTE-23 demonstrated marked polyspecificity and reacted with cytoskeletal proteins (vimentin, keratin, actin), single-stranded DNA, specific synthetic polynucleotides, and cardiolipin. Antibody RTE-23 also showed a lupus anticoagulant-like activity. Screening of sera of autoimmune disease patients with antinuclear antibodies revealed two patients, both with SLE, whose sera blocked antibody RTE-23 binding to nuclei and recognized nuclear proteins identical to those recognized by antibody RTE-23. These results suggested that antibody RTE-23 displays a pattern of self-antigen binding that is represented as well in SLE patient sera.

Authors
Laster, AJ; Pisetsky, DS; Haynes, BF
MLA Citation
Laster, AJ, Pisetsky, DS, and Haynes, BF. "Polyspecific reactivity of a murine monoclonal antibody that binds to nuclear matrix-associated, chromatin-bound autoantigens." Clin Immunol Immunopathol 44.2 (August 1987): 187-205.
PMID
3301102
Source
pubmed
Published In
Clinical Immunology and Immunopathology
Volume
44
Issue
2
Publish Date
1987
Start Page
187
End Page
205

Cyclosporine inhibition of CH series murine B-cell lymphomas.

To investigate the mechanisms by which cyclosporine (CsA) inhibits B-cell function, the effect of this agent on murine B-lymphoma cell lines of the CH series was tested. These lymphomas appear to be derived from a restricted B-cell population on the basis of their common expression of the Ly-1 cell surface marker and autoantibody products. Proliferation of each of the six cell lines tested was inhibited by CsA at doses without effect on the nonlymphoid HeLa cell line. The cell lines, however, differed from each other in their sensitivity to this agent. To correlate this sensitivity to other functional B-cell properties, the effect of lipopolysaccharide (LPS) on the proliferation of the CH cell lines was tested. Three of the lines showed enhanced proliferation to LPS; two were inhibited while one was unaffected. The cell lines that responded with increased proliferation to LPS were the most sensitive to CsA. These results indicate that sensitivity to CsA may be a common property of B cells of certain lineages, although the degree of sensitivity may be influenced by the activation properties of these cells.

Authors
Gorelick, MH; Bishop, GA; Haughton, G; Pisetsky, DS
MLA Citation
Gorelick, MH, Bishop, GA, Haughton, G, and Pisetsky, DS. "Cyclosporine inhibition of CH series murine B-cell lymphomas." Cell Immunol 107.1 (June 1987): 219-226.
PMID
3495347
Source
pubmed
Published In
Cellular Immunology
Volume
107
Issue
1
Publish Date
1987
Start Page
219
End Page
226

Characterization of monoclonal antibodies with specificity for DNA and the synthetic polypeptide antigen (T,G)-A-L.

Because of evidence for structural similarity of variable region genes of anti-DNA and anti-(T,G)-A-L antibodies, polyspecific interactions of monoclonal anti-DNA and anti-(T,G)-A-L antibodies were investigated. Of 20 monoclonal antibodies from C57BL/10 mice with (T,G)-A-L binding, two bound DNA as determined by ELISA. In contrast, two of five anti-DNA monoclonal antibodies from MRL-lpr/lpr mice bound (T,G)-A-L. For both sets of antibodies, antigen binding was shown to be the activity of the same antibody by cross-inhibition studies. To determine whether such polyspecific antibodies were expressed during autoimmune disease, sera of autoimmune MRL-lpr/lpr mice were tested for anti-(T,G)-A-L activity. This analysis demonstrated minimal elevations of anti-(T,G)-A-L in comparison to BALB/c controls. These studies thus confirm predictions about the binding activity of anti-(T,G)-A-L and anti-DNA antibodies based on structural analysis of variable region genes. They further indicate, that while anti-(T,G)-A-L and anti-DNA antibodies may have overlapping specificity, polyspecific antibodies of this kind are not preferentially expressed during autoimmunity.

Authors
Pisetsky, DS; Klatt, C; Pincus, SH
MLA Citation
Pisetsky, DS, Klatt, C, and Pincus, SH. "Characterization of monoclonal antibodies with specificity for DNA and the synthetic polypeptide antigen (T,G)-A-L." Mol Immunol 24.4 (April 1987): 365-369.
PMID
3498882
Source
pubmed
Published In
Molecular Immunology
Volume
24
Issue
4
Publish Date
1987
Start Page
365
End Page
369

Specificity analysis of monoclonal anti-DNA antibodies.

The specificity of a panel of murine monoclonal anti-DNA antibodies for DNA antigenic determinants was evaluated by testing their relative binding to various animal and bacterial DNAs. The antibody panel consisted of six monoclonal anti-DNAs of MRL-lpr/lpr and B6-lpr/lpr origin, while the antigens tested were calf thymus (CT), salmon testes (ST), E. coli (EC) and Micrococcus (MC) DNA. While all antibodies bound to CT, ST, and EC DNA to a similar extent by direct ELISA, only one showed an equivalent level of interaction with MC DNA. The relationship of antigenic sites recognized by the antibodies was evaluated further by competition ELISA, assessing the ability of the anti-DNAs to block the interaction of a biotinylated anti-DNA with solid-phase DNA antigen. For each of the DNAs tested, two patterns of DNA interaction could be distinguished on the basis of the relative inhibitory activity of the different monoclonals. These results suggest that anti-DNA antibodies can be characterized using naturally occurring DNAs, with the observed patterns of binding suggesting recognition of unique antigenic sites, some of which are discrete and non-overlapping.

Authors
Karounos, DG; Pisetsky, DS
MLA Citation
Karounos, DG, and Pisetsky, DS. "Specificity analysis of monoclonal anti-DNA antibodies." Immunology 60.4 (April 1987): 497-501.
PMID
3495481
Source
pubmed
Published In
Immunology
Volume
60
Issue
4
Publish Date
1987
Start Page
497
End Page
501

Functional alterations of macrophages in autoimmune MRL-lpr/lpr mice.

To assess the role of macrophages (MAC) in the pathogenesis of systemic lupus erythematosus, we investigated functional aspects of peritoneal MAC obtained from autoimmune MRL/MpJ-lpr/lpr (MRL-lpr) mice. MRL-lpr and control C3H/HeN MAC were obtained from untreated mice or mice injected i.p. with 1 ml of 10% sterile peptone 3 days before cell harvest. MRL-lpr mice had significantly more peritoneal cells (MAC and lymphocytes) than did control mice. In endotoxin-free conditions, MRL-lpr MAC were similar to C3H/HeN MAC in their baseline, and IFN-gamma and/or LPS enhanced cytolysis of 3T12 fibrosarcoma tumor cells. Compared with C3H/HeN MAC, MRL-lpr MAC had a significant increase in antibody-dependent cellular cytotoxicity activity against sheep erythrocytes. This enhanced activity was not accompanied by a similar increase in adherence and/or phagocytosis of the same targets. Finally, in response to phorbol myristate acetate stimulation, both resident and peptone-induced MAC from MRL-lpr mice produced significantly more hydrogen peroxide than did those from control mice. These results indicate that MAC from MRL-lpr mice display features of selective "activation", and suggest that MAC or their products may play a role in the pathogenesis of inflammatory disorders seen in autoimmune diseases.

Authors
Dang-Vu, AP; Pisetsky, DS; Weinberg, JB
MLA Citation
Dang-Vu, AP, Pisetsky, DS, and Weinberg, JB. "Functional alterations of macrophages in autoimmune MRL-lpr/lpr mice." J Immunol 138.6 (March 15, 1987): 1757-1761.
PMID
3102599
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
138
Issue
6
Publish Date
1987
Start Page
1757
End Page
1761

A new method for the cytological analysis of autoantibody specificities using whole-mount, surface-spread meiotic nuclei

A new method for the cytological analysis of antinuclear antibody binding offers several advantages over conventional techniques. Nuclei in meiosis, prepared by surface-spreading spermatocytes, provide a detailed examination of the constituents of the nucleus - euchromatin, heterochromatin, sex chromatin, nucleoli, centromeres, and dense patches that seem related to RNA metabolism - and each of these structures can be seen to change in morphology and antibody labeling during the course of meiotic prophase. Results using sera from humans and mice with autoimmune disease, and using several mouse monoclonal antibodies, demonstrate the potential of this method for clinical and research applications, both for more common antibody types and for those that bind epitopes which are unique to germ cells. © 1987.

Authors
Dresser, M; Pisetsky, D; Warren, R; McCarty, G; Moses, M
MLA Citation
Dresser, M, Pisetsky, D, Warren, R, McCarty, G, and Moses, M. "A new method for the cytological analysis of autoantibody specificities using whole-mount, surface-spread meiotic nuclei." Journal of Immunological Methods 104.1-2 (1987): 111-121.
PMID
3316391
Source
scival
Published In
Journal of Immunological Methods
Volume
104
Issue
1-2
Publish Date
1987
Start Page
111
End Page
121

IgG antinuclear antibodies with cross-reactive rheumatoid factor activity.

To investigate whether IgG antinuclear antibodies have cross-reactive rheumatoid factor activity, monoclonal IgG antibodies to DNA and Sm from autoimmune MRL-lpr/lpr mice were assayed by ELISA for binding to IgG antigens. Of the nine anti-DNA and anti-Sm monoclonals tested, six showed significant binding to affinity-purified rabbit IgG (RIgG) and human IgG (HIgG). To confirm that cross-reactivities were due to a single antibody, immunoabsorption of a representative polyspecific monoclonal termed C11 (anti-DNA, anti-Sm) on either Sepharose-DNA or Sepharose-RIgG resulted in marked loss of activity to the three antigens DNA, Sm and RIgG compared with immunoabsorption on Sepharose-bovine serum albumin. The monomolecular nature of the cross-reacting antibody was also suggested by inhibition analysis of C11; DNA inhibited C11 binding to RIgG 64%, whereas Sm inhibited binding to RIgG 33%. Aggregated RIgG and HIgG, however, did not inhibit binding of C11 to DNA, Sm, or solid-phase RIgG, probably reflecting the low affinity of this antibody for fluid phase Ig. Together, these findings suggest that antinuclear autoantibodies of the IgG, as well as the IgM, class have polyspecific IgG binding activity and suggest that IgG antinuclear antibodies may emerge from rheumatoid factor responses.

Authors
Darwin, BS; Grudier, JP; Klatt, CL; Pisetsky, DS
MLA Citation
Darwin, BS, Grudier, JP, Klatt, CL, and Pisetsky, DS. "IgG antinuclear antibodies with cross-reactive rheumatoid factor activity." J Immunol 137.12 (December 15, 1986): 3796-3801.
PMID
3782796
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
137
Issue
12
Publish Date
1986
Start Page
3796
End Page
3801

Antibodies to DNA. A perspective.

Authors
Emlen, W; Pisetsky, DS; Taylor, RP
MLA Citation
Emlen, W, Pisetsky, DS, and Taylor, RP. "Antibodies to DNA. A perspective." Arthritis Rheum 29.12 (December 1986): 1417-1426. (Review)
PMID
3541943
Source
pubmed
Published In
Arthritis and Rheumatism
Volume
29
Issue
12
Publish Date
1986
Start Page
1417
End Page
1426

The intravenous insulin tolerance test in type I diabetes.

The intravenous insulin tolerance test (ITT) allows general assessment of insulin sensitivity by determining the fall in plasma glucose after injection of 0.1 unit/kg regular insulin. To evaluate the usefulness of this test diagnostically in distinguishing Type I from Type II diabetes, 24 patients with Type I diabetes (defined as early age of onset, ideal body weight or less at onset, and insulin dependence) underwent ITTs. Seven patients had normal glucose disposal rates (Ki greater than or equal to 2.5. Nine patients had Ki less than 2.5 but greater than 1.0. Eight patients had Ki less than 1.0. The slopes did not correlate with the control of the diabetes (as assessed by measurement of glycosylated hemoglobin), the presence or titer of anti-insulin antibodies, the duration of the diabetes, the age of onset, the presence of complications, or the current insulin dose. These results indicate that varying amounts of insulin resistance may be present in Type I diabetes and cannot necessarily be explained by poor control or the presence of insulin antibodies.

Authors
Lucas, KJ; Karounos, DG; Ellis, GJ; Morris, MA; Pisetsky, DS; Feinglos, MN
MLA Citation
Lucas, KJ, Karounos, DG, Ellis, GJ, Morris, MA, Pisetsky, DS, and Feinglos, MN. "The intravenous insulin tolerance test in type I diabetes." Res Commun Chem Pathol Pharmacol 53.3 (September 1986): 331-345.
PMID
3534980
Source
pubmed
Published In
Research Communications in Chemical Pathology and Pharmacology
Volume
53
Issue
3
Publish Date
1986
Start Page
331
End Page
345

The clearance of a monoclonal anti-DNA antibody following administration of DNA in normal and autoimmune mice.

To study the assembly of DNA-anti-DNA complexes in vivo, we have measured the clearance from blood and organ localization of a murine IgG2a monoclonal anti-DNA antibody, called 6/0, following the infusion of DNA intravenously or intraperitoneally. Intraperitoneal DNA caused a profound acceleration of 6/0 anti-DNA clearance that was dose dependent and demonstrable after the infusion of as little as 1.9 microgram per gram of body weight of single-stranded DNA. The antibody was cleared primarily in the liver without increased deposition in the kidney. Intraperitoneal infusions of DNA also accelerated the clearance of 6/0 in autoimmune MRL-lpr/lpr mice. In contrast, intravenous DNA given in comparable doses caused only a slight increase in 6/0 antibody clearance; this accelerated clearance was seen only at low antigen doses and only during the first 10 min following DNA infusion. Using double-radiolabeling techniques, 6/0 and Cl.18, an IgG2ak myeloma protein without anti-DNA activity, were found to disappear from blood at a comparable rate in both B6D2 mice and MRL-lpr/lpr mice. These results suggest that the DNA-anti-DNA immune complexes can form in vivo but that this process is profoundly affected by the manner in which DNA enters the circulation. In addition, the results suggest that DNA-dependent clearance is not a major pathway for anti-DNA metabolism in normal or at least one strain of autoimmune mice.

Authors
Jones, FS; Pisetsky, DS; Kurlander, RJ
MLA Citation
Jones, FS, Pisetsky, DS, and Kurlander, RJ. "The clearance of a monoclonal anti-DNA antibody following administration of DNA in normal and autoimmune mice." Clin Immunol Immunopathol 39.1 (April 1986): 49-60.
PMID
3948437
Source
pubmed
Published In
Clinical Immunology and Immunopathology
Volume
39
Issue
1
Publish Date
1986
Start Page
49
End Page
60

Cyclosporine inhibition of a murine B cell lymphoma.

The effect of cyclosporine (CsA) on the CH12 murine B cell lymphoma was investigated to determine whether sensitivity to this agent is retained by malignant B cells. This tumour produces an antibody to bromelain-treated red blood cells and may represent transformation of a B cell with certain activation properties associated with early resting B cells. In in vitro cultures, the growth and proliferation of CH12 were inhibited by CsA in concentrations of 0.1-1.0 microgram/ml; these levels were ineffective against non-lymphoid tumours, although some non-specific cell toxicity was noted at higher levels. IgM antibody production, as measured by enzyme-linked immunosorbent assay (ELISA), was inhibited over the same range. CH12 cells stimulated by lipopolysaccharide, however, were less sensitive to CsA than untreated cells. These studies indicate that malignant B cells may be sensitive to CsA, perhaps reflecting their derivation from a functionally distinct B cell population with enhanced drug sensitivity.

Authors
Pisetsky, DS; Haughton, G
MLA Citation
Pisetsky, DS, and Haughton, G. "Cyclosporine inhibition of a murine B cell lymphoma." Clin Exp Immunol 63.3 (March 1986): 549-554.
PMID
3486733
Source
pubmed
Published In
Clinical & Experimental Immunology
Volume
63
Issue
3
Publish Date
1986
Start Page
549
End Page
554

Systemic lupus erythematosus.

Systemic lupus erythematosus is a multisystem inflammatory disease characterized by autoantibody production. Recent investigations are providing insights into the immunoregulatory disturbances underlying this disease, and are clarifying the approach to diagnosis and treatment.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Systemic lupus erythematosus." Med Clin North Am 70.2 (March 1986): 337-353. (Review)
PMID
3512930
Source
pubmed
Published In
Medical Clinics of North America
Volume
70
Issue
2
Publish Date
1986
Start Page
337
End Page
353

Specificity and idiotypic analysis of a monoclonal anti-Sm antibody with anti-DNA activity.

To investigate the mechanisms of anti-Sm expression in murine systemic lupus erythematosus (SLE), the idiotypic determinants of a monoclonal anti-Sm antibody were studied. This antibody, 2G7, was derived from the fusion of spleen cells of an autoimmune MRL-lpr/lpr mouse with the 653 myeloma. Specificity for Sm was demonstrated by an ELISA with the use of affinity-purified Sm as well as immunoprecipitation of radiolabeled RNA. An anti-2G7 antiserum was prepared in a rabbit and rendered specific for idiotype by extensive absorption against BALB/c and MRL-lpr/lpr monoclonal proteins. The resulting antiserum detected determinants found on 2G7 as well as on two MRL anti-DNA antibodies, 6/P and 6/N, an independently derived MRL-lpr/lpr anti-Sm called 7.13, and the BALB/c myeloma FLOPC 21. Two distinct determinants could be demonstrated by the creation of cross-reactive idiotype systems by using the various monoclonal antibodies as ligands for the anti-idiotype. Both idiotypes were demonstrated in sera of normal and autoimmune mice, although MRL +/-/+/- mice had the highest levels of strains tested. To explain the pattern of idiotypic relatedness, 2G7 was tested for anti-DNA activity by ELISA. 2G7 displayed activity for single-stranded DNA as well as synthetic DNA and RNA homopolymers. Absorption analysis indicated that the anti-DNA and anti-Sm binding activities were the product of the same antibody. These results suggest that anti-Sm and anti-DNA may be related by both idiotype and antigenic specificity, providing a mechanism for their common expression in SLE.

Authors
Pisetsky, DS; Hoch, SO; Klatt, CL; O'Donnell, MA; Keene, JD
MLA Citation
Pisetsky, DS, Hoch, SO, Klatt, CL, O'Donnell, MA, and Keene, JD. "Specificity and idiotypic analysis of a monoclonal anti-Sm antibody with anti-DNA activity." J Immunol 135.6 (December 1985): 4080-4085.
PMID
3877763
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
135
Issue
6
Publish Date
1985
Start Page
4080
End Page
4085

Anti-Sm autoantibodies in MRL mice: analysis of precursor frequency.

Individual MRL-lpr mice vary in their capacity to generate anti-Sm autoantibodies spontaneously. We have compared the frequency of B-cell precursors for this autoantibody in serologically negative and serologically positive MRL-lpr mice, and in normals. Anti-Sm precursors were present in a frequency of approximately 1 per 10-30,000 in spleen cell cultures from anti-Sm positive mice, but were undetectable when spleen cells from serologically negative MRL-lpr mice or from normal mice were examined. Despite LPS stimulation, neither IgM nor IgG precursors could be detected. In parallel cultures, in contrast, anti-DNA autoantibody precursors were readily detected. The results thus indicate that, for the lupus-specific autoantibodies, the absence of antibody in autoimmune mice reflects a deficit in precursor B lymphocytes rather than an active regulatory mechanism. It is suggested that the generation of anti-Sm may reflect a low-probability random event in the generation of B-cell diversity.

Authors
Cohen, PL; Shores, EW; Rapoport, R; Caster, S; Eisenberg, RA; Pisetsky, DS
MLA Citation
Cohen, PL, Shores, EW, Rapoport, R, Caster, S, Eisenberg, RA, and Pisetsky, DS. "Anti-Sm autoantibodies in MRL mice: analysis of precursor frequency." Cell Immunol 96.2 (December 1985): 448-454.
PMID
3879807
Source
pubmed
Published In
Cellular Immunology
Volume
96
Issue
2
Publish Date
1985
Start Page
448
End Page
454

The influence of Yaa on anti-DNA responses of B6-lpr mice.

The anti-DNA autoantibody responses of mice congenic for lpr and the Y-linked autoimmune accelerator (Yaa) genes were studied to evaluate genetic interactions in murine autoimmunity. Male B6-lpr, + mice failed to generate significant anti-DNA responses in comparison to B6-+, + mice. In contrast, B6-lpr mice bearing Yaa (B6-lpr, Yaa) had markedly increased IgG anti-DNA levels in comparison to both B6-+, + and B6-lpr, + mice. To determine whether anti-DNA levels reflected the overall B-cell response to lpr and Yaa, total IgG and IgM levels were also determined. This analysis demonstrated that the increase in IgG anti-DNA produced by mice with the Yaa gene was far greater than the increase in total IgG. Taken together, these results indicate that an impaired anti-DNA response related to one gene-determined mechanism for the development of autoimmunity does not preclude the response to another. Furthermore, it appears that the polyclonal B-cell activation during murine autoimmunity may be associated with the preferential expression of certain autoantibodies.

Authors
Pisetsky, DS; Klatt, C; Dawson, D; Roths, JB
MLA Citation
Pisetsky, DS, Klatt, C, Dawson, D, and Roths, JB. "The influence of Yaa on anti-DNA responses of B6-lpr mice." Clin Immunol Immunopathol 37.3 (December 1985): 369-376.
PMID
3931946
Source
pubmed
Published In
Clinical Immunology and Immunopathology
Volume
37
Issue
3
Publish Date
1985
Start Page
369
End Page
376

Inhibition of in vitro anti-DNA B-cell responses by cyclosporine.

The action of the immunosuppressive agent cyclosporine (CsA) on anti-DNA B-cell responses was investigated in an in vitro system. Spleen cells from autoimmune MRL-lpr/lpr or control BALB/c mice, when cultured at high cell density, spontaneously produced significant amounts of IgM and IgG, including anti-DNA. IgG levels, both total and anti-DNA, were much higher for MRL-lpr/lpr cells compared to BALB/c cells, suggesting similarity of this model system to the in vivo response. For cells of both strains, the production of IgM and IgG anti-DNA was 10- to 100-fold more sensitive to the inhibitory activity of CsA than total immunoglobulin production. The effect was not manifest, however, in cultures stimulated with the B-cell mitogen, lipopolysaccharide. These observations suggest that CsA in certain dose ranges preferentially inhibits anti-DNA production, with efficacy determined by the mechanisms promoting B-cell hyperactivity.

Authors
Mayus, JL; Semper, KF; Pisetsky, DS
MLA Citation
Mayus, JL, Semper, KF, and Pisetsky, DS. "Inhibition of in vitro anti-DNA B-cell responses by cyclosporine." Cell Immunol 94.1 (August 1985): 195-204.
PMID
3874703
Source
pubmed
Published In
Cellular Immunology
Volume
94
Issue
1
Publish Date
1985
Start Page
195
End Page
204

Defects in mononuclear phagocytic system (MPS) function in autoimmune MRL-lpr/lpr mice.

MRL-lpr/lpr mice develop an autoimmune disease similar to systemic lupus erythematosus. To determine whether mice of this strain develop defects in mononuclear phagocyte system (MPS) function similar to those observed in patients, the pattern of sequestration of labeled immune complexes was compared 90 min after infusion into MRL-lpr/lpr and into normal B6D2 mice. The amount of complexes persisting in the blood was increased, and the amount sequestered in the liver was significantly reduced in MRL-lpr/lpr mice in comparison to normal B6D2 controls. This defect was most evident in MRL-lpr/lpr mice of the ages of 25-26 weeks; mice of this age also demonstrated the greatest elevation of anti-DNA antibody levels. The role of the MRL strain background and of the lpr gene in determining this defect was investigated by analysis of MRL-+/-/+/- and of other lpr congenic strains (B6-lpr/lpr, AKR-lpr/lpr, and C3H-lpr/lpr). Both MRL-+/-/+/- and congenic lpr animals showed similar defects, although to a lesser degree than MRL-lpr/lpr mice. In contrast, MRL-lpr/lpr mice demonstrated normal clearance of heat-damaged red blood cells and heat-aggregated albumin. Thus MRL-lpr/lpr mice display a selective defect in MPS Fc receptor function and may provide a valuable model for elucidating the etiology and importance of MPS dysfunction in immune complex deposition disease.

Authors
Jones, FS; Pisetsky, DS; Kurlander, RJ
MLA Citation
Jones, FS, Pisetsky, DS, and Kurlander, RJ. "Defects in mononuclear phagocytic system (MPS) function in autoimmune MRL-lpr/lpr mice." Clin Immunol Immunopathol 36.1 (July 1985): 30-39.
PMID
3874027
Source
pubmed
Published In
Clinical Immunology and Immunopathology
Volume
36
Issue
1
Publish Date
1985
Start Page
30
End Page
39

Expression of a highly conserved anti-DNA idiotype in normal and autoimmune mice.

The specificity and idiotypic relationships of a monoclonal anti-DNA antibody were investigated to evaluate genetic control in this autoantibody response. 6/0 is an IgG2a monoclonal anti-DNA derived by the fusion of spleen cells from an autoimmune MRL-lpr/lpr mouse and the cell line NS1. By an enzyme-linked immunosorbent assay (ELISA) for anti-DNA, 6/0 demonstrated preference for single-stranded DNA and bound deoxyribo- and ribohomopolymers of dissimilar base composition. The control of 6/0 expression was evaluated by idiotypic analysis using a rabbit anti-6/0 antiserum made specific by absorption with the BALB/c myelomas UPC 10 (IgG2a) and MOPC 21 (IgG1). The resulting preparation was fractionated by BALB/c IgG affinity columns to provide antibodies to idiotypic determinants essentially unique to 6/0 and those commonly expressed in sera. The commonly expressed 6/0 idiotype was found in sera of ten inbred strains of mice and was not exclusive to the autoimmune strains. MRL-lpr/lpr and A/J strain mice displayed idiotype levels almost fivefold greater than other strains, with 6/0 idiotype-bearing antibodies having serum concentrations as high as 1 mg/ml. Levels of the 6/0 idiotype, however, did not correlate with anti-DNA levels among the various strains. In addition to mice, the majority of individuals of three inbred rat strains showed detectable 6/0 idiotype in their sera. These results suggest that the 6/0 idiotype, although identified using a monoclonal anti-DNA antibody, represents a framework determinant that is phylogenetically conserved. The mechanisms for the expression of this determinant may differ among the normal and autoimmune strains.

Authors
Mayus, JL; Pisetsky, DS
MLA Citation
Mayus, JL, and Pisetsky, DS. "Expression of a highly conserved anti-DNA idiotype in normal and autoimmune mice." Clin Immunol Immunopathol 34.3 (March 1985): 366-378.
PMID
2578910
Source
pubmed
Published In
Clinical Immunology and Immunopathology
Volume
34
Issue
3
Publish Date
1985
Start Page
366
End Page
378

Hyper-Ia antigen expression on B cells from B6-lpr/lpr mice correlates with manifestations of the autoimmune state.

To investigate the state of activation of B cells from mice with the lpr gene defect, membrane Ia antigen (mIa) expression was analyzed on B cells from B6-lpr/lpr (lpr) and control B6- +/-/+/- mice. B cells from lpr mice exhibited marked increases in levels of mIa as determined by flow cytometry using a monoclonal anti-I-Ab,d reagent. This increase, which was progressive with age, suggests that phenotypic alteration of B-cell mIa expression is a consequence of lpr gene action. Since B-cell activation manifest by elevated mIa expression may promote productive interactions with helper T cells, these observations suggest an important role for B-cell abnormalities in the etiology of lpr-induced autoimmune disease.

Authors
Monroe, JG; Cambier, JC; Mody, EA; Pisetsky, DS
MLA Citation
Monroe, JG, Cambier, JC, Mody, EA, and Pisetsky, DS. "Hyper-Ia antigen expression on B cells from B6-lpr/lpr mice correlates with manifestations of the autoimmune state." Clin Immunol Immunopathol 34.1 (January 1985): 124-129.
PMID
3871181
Source
pubmed
Published In
Clinical Immunology and Immunopathology
Volume
34
Issue
1
Publish Date
1985
Start Page
124
End Page
129

Antinuclear antibodies and nuclear antigens in NZB myeloma ascitic fluids

Approximately 5-10% of ascitic fluids from 411 NZB myeloma tumors were found to possess either antinuclear (ANA) or Coombs antibodies. Some fluids showed anti-Sm specificity, which is thought to be unique to systemic lupus erythematosus (SLE). In addition, the ascitic fluids were found to contain large amounts of DNA and Sm autoantigens. Further passage of ANA-positive myelomas indicated that the autoantibodies were not products of the myeloma cells themselves. The sporadic appearance of unsuspected autoantibodies in ascitic fluids may cause confusion when working with myeloma or with hybridoma reagents. Furthermore, the development of SLE-specific anti-Sm antibodies in this context suggests parallels between myeloma development and autoimmunity. © 1985.

Authors
Eisenberg, RA; Pisetsky, D; Cohen, PL
MLA Citation
Eisenberg, RA, Pisetsky, D, and Cohen, PL. "Antinuclear antibodies and nuclear antigens in NZB myeloma ascitic fluids." Clinical Immunology and Immunopathology 35.3 (1985): 337-345.
PMID
3872756
Source
scival
Published In
Clinical Immunology and Immunopathology
Volume
35
Issue
3
Publish Date
1985
Start Page
337
End Page
345

Monoclonal rheumatoid factors from B6-lpr/lpr mice.

Monoclonal rheumatoid factors (MoRF) were prepared from autoimmune B6-lpr/lpr mice to investigate the influence of strain background on the specificity of these autoantibodies. Using screening assays for binding to heterologous rabbit IgG, four IgM MoRF were obtained. Three of these antibodies showed a broad pattern of reactivity with IgG antigen, binding BALB/c myeloma IgG1, IgG2a and IgG3 as well as heterologous IgG. One of the antibodies, however, had a distinct form of IgG interaction and was without reactivity with any of the BALB/c myelomas tested. None of the antibodies had significant reactivity with IgG2b. These results suggest common features of B6-lpr/lpr rheumatoid factor (RF) specificities, some of which may be shared by comparable products derived from MRL-lpr/lpr mice. Comparison of these antibodies with those in other reported series suggests that the background genome as well as the nature of the inducing mechanisms may affect the specificity of RF as well as their pathogenetic role.

Authors
Warren, RW; Sailstad, DM; Pisetsky, DS
MLA Citation
Warren, RW, Sailstad, DM, and Pisetsky, DS. "Monoclonal rheumatoid factors from B6-lpr/lpr mice." Clin Exp Immunol 58.3 (December 1984): 731-736.
PMID
6509801
Source
pubmed
Published In
Clinical & Experimental Immunology
Volume
58
Issue
3
Publish Date
1984
Start Page
731
End Page
736

Influence of assay conditions on ELISA determinations of anti-DNA antibodies.

The influence of assay conditions on anti-DNA determinations by an enzyme-linked immunosorbent assay (ELISA) was investigated to evaluate the detection of various DNA antigenic specificities. Among 4 monoclonal anti-DNA antibodies of MRL-lpr/lpr strain origin, 2 showed higher titers in 100 mM NaCl-50 mM Tris, pH 7.5 (Tris-NaCl) than in phosphate-buffered saline (PBS). The determination of 'polyspecificity' for these monoclonal products also depended on the set of conditions used for assay with inhibitory activity of polynucleotides differing in the 2 buffers. The buffer effects were not confined to the monoclonal antibodies as increases in anti-DNA titers were demonstrated for certain SLE patient sera when assayed in Tris-NaCl rather than PBS; sera of MRL-lpr/lpr mice showed an opposite effect, however, with enhancement of anti-DNA activity by PBS. These results suggest that the representation of antigenic sites on DNA may be variably affected by the conditions of assay, altering quantitative and qualitative assessment of this important serological marker.

Authors
Pisetsky, DS; Semper, KF
MLA Citation
Pisetsky, DS, and Semper, KF. "Influence of assay conditions on ELISA determinations of anti-DNA antibodies." J Immunol Methods 74.2 (November 30, 1984): 217-227.
PMID
6209337
Source
pubmed
Published In
Journal of Immunological Methods
Volume
74
Issue
2
Publish Date
1984
Start Page
217
End Page
227

Idiotypic analysis of a monoclonal anti-Sm antibody. II. Strain distribution of a common idiotypic determinant and its relationship to anti-Sm expression.

The idiotypes borne by Y2, a monoclonal anti-Sm antibody of MRL-lpr/lpr mouse strain origin, were investigated to elucidate genetic mechanisms in this autoantibody response. An anti-Y2 anti-idiotypic antiserum was raised in a rabbit and was rendered specific for idiotype by extensive absorption with globulins of the B6 and BALB/c strains as well as the BALB/c myeloma UPC 10. By using a sensitive assay for idiotype by inhibition ELISA, the Y2 determinant was found to be commonly expressed in sera of MRL-lpr/lpr and MRL-+/+ mice. Moreover, sera of several normal strain mice also bore the idiotype and, in mice bearing the lpr gene, idiotype levels were increased 1.5 to fivefold, even in the absence of a serum anti-Sm response. The relationship of this idiotype to anti-Sm expression was further assessed by determining the idiotype content of affinity-purified anti-Sm antibodies from MRL-lpr/lpr mice. Anti-Sm from serum pools or individual animals showed no significant enrichment of the Y2 idiotype in comparison to unselected MRL-lpr/lpr IgG. These results suggest that the Y2 idiotype defines only a minor component of the anti-Sm autoantibody response, and that most antibodies with this determinant express other antigenic specificities.

Authors
Pisetsky, DS; Semper, KF; Eisenberg, RA
MLA Citation
Pisetsky, DS, Semper, KF, and Eisenberg, RA. "Idiotypic analysis of a monoclonal anti-Sm antibody. II. Strain distribution of a common idiotypic determinant and its relationship to anti-Sm expression." J Immunol 133.4 (October 1984): 2085-2089.
PMID
6206150
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
133
Issue
4
Publish Date
1984
Start Page
2085
End Page
2089

Characterization and idiotypic analysis of an anti-RNP monoclonal antibody.

To investigate mechanisms of anti-RNP antibody expression in autoimmune disease, idiotypes of a monoclonal anti-RNP of murine origin were analysed. This antibody, designated 4L1, was obtained from a MRL-lpr/lpr mouse and shown to have anti-RNP specificity by gel analysis of radiolabelled cellular RNA. An anti-idiotypic antiserum was prepared in a rabbit to 4L1 and rendered specific for idiotype by absorption with IgG from B6 mice and two BALB/c myelomas of the same chain composition as 4L1 (IgM kappa). In competition ELISA assays, this antiserum detected idiotypes commonly expressed in sera of MRL-lpr/lpr mice irrespective of the presence of anti-RNP. These idiotypes were not exclusive to this autoimmune strain, however, and could also be identified in normal mice. To identify other antibodies with this idiotype, a panel of MRL hybridomas was tested. This analysis demonstrated idiotypic cross-reactivity between 4L1 and two anti-Sm monoclonal antibodies derived from another animal. These results suggest that 4L1 belongs to a larger idiotype bearing family only some of whose members may have aberrant expression.

Authors
Pisetsky, DS; Salistad, DM; Chambers, JC
MLA Citation
Pisetsky, DS, Salistad, DM, and Chambers, JC. "Characterization and idiotypic analysis of an anti-RNP monoclonal antibody." Clin Exp Immunol 56.3 (June 1984): 593-600.
PMID
6744662
Source
pubmed
Published In
Clinical & Experimental Immunology
Volume
56
Issue
3
Publish Date
1984
Start Page
593
End Page
600

Specificity analysis of monoclonal anti-DNA antibodies from B6-lpr/lpr mice.

The binding properties of B6-lpr/lpr anti-DNA monoclonal antibodies were characterized to evaluate the influence of genetic background on the diversity and specificity of lpr-induced autoantibody responses. Six anti-DNA antibodies were produced from fusions with B6-lpr/lpr mice, while another was obtained from a fusion with a B6-+/+ mouse immunized with lipopolysaccharide. Each antibody bound single-stranded DNA in preference to double-stranded DNA, with variation of over 300-fold in relative binding activities. In terms of binding to a panel of synthetic polynucleotides, each antibody exhibited a unique antigenic specificity. This binding, however, was not dependent on recognition of a unique base or sugar moiety, since individual antibodies bound polymers of dissimilar composition. These results suggest a diversity of binding reactions for B6-lpr/lpr anti-DNA antibodies, with a clonal repertoire similar to that of mice from autoimmune backgrounds.

Authors
Warren, RW; Sailstad, DM; Caster, SA; Pisetsky, DS
MLA Citation
Warren, RW, Sailstad, DM, Caster, SA, and Pisetsky, DS. "Specificity analysis of monoclonal anti-DNA antibodies from B6-lpr/lpr mice." Arthritis Rheum 27.5 (May 1984): 545-551.
PMID
6721887
Source
pubmed
Published In
Arthritis and Rheumatism
Volume
27
Issue
5
Publish Date
1984
Start Page
545
End Page
551

The influence of the lpr gene on B cell activation: differential antibody expression in lpr congenic mouse strains.

Spontaneous immunoglobulin production in four strains of lpr/lpr congenic mice was investigated to identify genetic interactions in lpr-induced polyclonal B cell activation. Sera were obtained from male and female lpr/lpr mice of the MRL, B6, C3H, and AKR strains as well as controls of +/+ genotypes. Antibody levels were compared at the time of peak response. Quantitative antibody determinations were performed by isotype-specific ELISA assays using responses to single-stranded DNA (sDNA), mouse IgG, rabbit IgG, and keyhole limpet hemocyanin as models. Among the strains studied there were significant differences in the antibody levels observed, with the strain producing highest levels dependent on the response measured. Thus, MRL-lpr/lpr produced the highest levels of IgG anti-sDNA while B6-lpr/lpr mice produced more anti-IgG than mice of other strains. Within each strain, the lpr gene appeared to affect the IgG more than the IgM response. A consistently high response by females was observed only in B6-lpr/lpr mice. These studies suggest that lpr-induced polyclonal B cell activation is influenced by the background genome with the extent of these genetic effects variable among responses.

Authors
Warren, RW; Caster, SA; Roths, JB; Murphy, ED; Pisetsky, DS
MLA Citation
Warren, RW, Caster, SA, Roths, JB, Murphy, ED, and Pisetsky, DS. "The influence of the lpr gene on B cell activation: differential antibody expression in lpr congenic mouse strains." Clin Immunol Immunopathol 31.1 (April 1984): 65-77.
PMID
6607806
Source
pubmed
Published In
Clinical Immunology and Immunopathology
Volume
31
Issue
1
Publish Date
1984
Start Page
65
End Page
77

Idiotypic cross-reaction between MRL monoclonal autoantibodies with different antigen specificity.

The idiotypic determinants borne by Y2, a monoclonal anti-SM antibody of MRL mouse strain origin, were investigated to elucidate mechanisms for the generation of autoantibodies. Using an ELISA assay, a rabbit anti-Y2 anti-idiotypic antiserum was tested for binding to other MRL autoantibodies to identify cross-reactive determinants. In this way, a shared idiotype was demonstrated for Y2 and 4K1, 4k1 is an IgG2bK monoclonal antibody derived from a fusion with spleen cells of an MRL-lpr/lpr mouse expressing anti-Sm. 4K1 displayed an antigenic specificity distinct from anti-Sm, however, with reactivity by fluorescent antinuclear antibody analysis to a subcellular component localized to the perinuclear region. The expression of the Y2-4K1 idiotype was assessed using a sensitive inhibition ELISA. The Y2-4K1 determinant was present in sera of MRL-lpr/lpr mice irrespective of the presence of anti-Sm. Sera of some but not all normal mouse strains also demonstrated this determinant. These results indicate that while some idiotypes may be associated with autoantibodies, they are not restricted to the autoimmune setting. This pattern of expression suggests that autoantibodies are part of larger idiotype-bearing antibody families with structural similarity to antibodies produced by normal mice. The autoimmune setting may promote the emergence of idiotype-bearing molecules with autoantibody activity and aberrant regulation.

Authors
Pisetsky, DS; Caster, SA
MLA Citation
Pisetsky, DS, and Caster, SA. "Idiotypic cross-reaction between MRL monoclonal autoantibodies with different antigen specificity." Clin Immunol Immunopathol 30.3 (March 1984): 461-468.
PMID
6199145
Source
pubmed
Published In
Clinical Immunology and Immunopathology
Volume
30
Issue
3
Publish Date
1984
Start Page
461
End Page
468

Mechanisms of polyclonal B-cell activation in autoimmune B6-lpr/lpr mice.

The influence of the lpr gene on spontaneous and lipopolysaccharide (LPS)-induced immunoglobulin production was studied in B6 mice homozygous for the mutant lpr gene (B6-lpr/lpr). Male and female mice of this congenic strain were followed for 1 year and sera serially tested by the enzyme-linked immunosorbent assay (ELISA) for the production of antibodies to single-stranded DNA (anti-sDNA), immunoglobulin (anti-IgG), and keyhole limpet hemocyanin (anti-KLH), models of autoantibody and non-autoantibody responses, respectively. Female B6-lpr/lpr mice demonstrated marked spontaneous responses to all three antigens; the responses of male B6-lpr/lpr mice were significantly lower but still exceeded those of the congenic B6-+/+ controls. These results demonstrate a generalized influence of sex on lpr associated responses. To determine whether this sex difference could be demonstrated with other forms of B-cell activation, young B6-+/+ and B6-lpr/lpr male and female mice were immunized with lipopolysaccharide and the induced responses determined. This immunization caused significant increases in the IgM response only. The levels of the induced responses produced after LPS treatment were comparable for +/+ and lpr/lpr mice. These results indicate that the enhanced responsiveness of female mice to lpr action is not reflected in the polyclonal response to LPS, which, furthermore, was unaffected by the presence of lpr. The differential influence of sex on lpr and LPS-induced responses and their apparent independence suggests that lpr and LPS promote B-cell activation by dissimilar mechanisms.

Authors
Warren, RW; Roths, JB; Murphy, ED; Pisetsky, DS
MLA Citation
Warren, RW, Roths, JB, Murphy, ED, and Pisetsky, DS. "Mechanisms of polyclonal B-cell activation in autoimmune B6-lpr/lpr mice." Cell Immunol 84.1 (March 1984): 22-31.
PMID
6199122
Source
pubmed
Published In
Cellular Immunology
Volume
84
Issue
1
Publish Date
1984
Start Page
22
End Page
31

Cloning of B cells from autoimmune MRL-lpr/lpr and MRL.xid mice.

The relationship between colony formation (cloning) of B cells and their activation in murine autoimmunity was investigated in MRL-lpr/lpr and MRL.xid mice. Cells from MRL-lpr/lpr mice showed similar requirements for in vitro growth as normal CBA/J and BALB/c cells, with maximal colony formation in the presence of the supporting factors lipopolysaccharide and sheep red blood cells. The frequency of colony-forming cells from MRL-lpr/lpr spleens or hapten-specific B-cell preparations was slightly greater than the two normal control strains, with this difference significant only for a comparison of BALB/c and MRL-lpr/lpr spleens. In contrast, MRL-lpr/lpr mice bearing the xid gene for B-cell immunodeficiency (MRL.xid) had markedly reduced B-cell colony formation. These mice nevertheless expressed anti-DNA antibodies, although at levels reduced from that of MRL-lpr/lpr controls. These results indicate that enhanced in vitro colony formation need not accompany B-cell hyperactivity in murine autoimmune disease and that autoantibody production can occur in mice with impairment in this growth property.

Authors
Pisetsky, DS; Caster, SA; Piper, M; Scott, DW; Steinberg, AD
MLA Citation
Pisetsky, DS, Caster, SA, Piper, M, Scott, DW, and Steinberg, AD. "Cloning of B cells from autoimmune MRL-lpr/lpr and MRL.xid mice." Cell Immunol 84.1 (March 1984): 32-40.
PMID
6607779
Source
pubmed
Published In
Cellular Immunology
Volume
84
Issue
1
Publish Date
1984
Start Page
32
End Page
40

Hybridoma SLE autoantibodies: insights for the pathogenesis of autoimmune disease.

Authors
Pisetsky, DS
MLA Citation
Pisetsky, DS. "Hybridoma SLE autoantibodies: insights for the pathogenesis of autoimmune disease." Clin Immunol Rev 3.2 (1984): 169-234. (Review)
PMID
6084573
Source
pubmed
Published In
Clinical immunology reviews
Volume
3
Issue
2
Publish Date
1984
Start Page
169
End Page
234

Effect os xid on anti-DNA B-cell precursors.

The influence of the xid gene on murine autoimmunity was investigated by precursor frequency analysis of anti-DNA-producing B cells in non-xid and congenic xid autoimmune and normal mice. Antibody responses were induced in vitro by lipopolysaccharides under limiting-dilution conditions with IgM anti-DNA and antitrinitrophenyl (TNP) determined as models of an autoantibody and nonautoantibody, respectively. All mice demonstrated high frequencies of precursors for anti-DNA without differences between the autoimmune (NZB and MRL-lpr/lpr) and nonautoimmune (CBA/J and DBA/2) strains. In these strains, the presence of the xid gene was associated with a marked reduction of anti-DNA. For NZB.xid mice, this loss of anti-DNA precursors was much greater than that for anti-TNP precursors, whose frequency was similar to that of NZB mice. In contrast, xid-bearing CBA/N, DBA/2.xid, and MRL.xid mice showed marked reductions in both anti-DNA and anti-TNP precursors relative to non-xid mice. The intact anti TNP response of NZB.xid mice may be an additional manifestation of immune abnormalities of the NZB strain and suggests that their B-cell populations differ from those of other mice in the conditions for activation. Since NZB.xid spleens had a preferential reduction of anti-DNA responses, this result suggests that in NZB mice precursors for anti-DNA antibodies are predominantly distributed in the B-cell subset affected by xid, explaining the abrogation of their autoantibody production. The equivalent number of precursors in non-xid mice indicates, however, that additional regulatory abnormalities underlie the change from precursor to autoantibody-producing B cell.

Authors
Pisetsky, DS; Caster, SA; Steinberg, AD
MLA Citation
Pisetsky, DS, Caster, SA, and Steinberg, AD. "Effect os xid on anti-DNA B-cell precursors." Cell Immunol 78.2 (June 1983): 326-332.
PMID
6602656
Source
pubmed
Published In
Cellular Immunology
Volume
78
Issue
2
Publish Date
1983
Start Page
326
End Page
332

Idiotypic cross-reaction between MRL autoantibodies and a BALB/c myeloma.

Y2, a monoclonal anti-Sm antibody of MRL origin, demonstrates an idiotype commonly expressed in autoimmune MRL mice, although not necessarily associated with anti-Sm activity. To identify non-anti-Sm antibodies with this common idiotype, a rabbit anti-Y2 anti-idiotypic antiserum was tested for activity with other MRL hybridoma products (HP) as well as BALB/c myelomas. Idiotypic cross-reactivity was thus demonstrated for Y2 and the product of the BALB/c fusing cell line 45.6TG1.7 (M45), an antibody without known antigen binding activity. This determinant was denoted the Y2-M45 idiotype and its presence in serum and other antibodies tested by an inhibition ELISA. The idiotype was demonstrated on some, but not all, MRL HP's tested and was not confined to antibodies of a unique specificity. The idiotype was also present in sera of some normal as well as autoimmune strains of mice with highest levels in two strains bearing the lpr gene. These results indicate that idiotypes of anti-Sm antibodies are not exclusive to the pathologic setting but may be found commonly expressed in mice with antibodies of a variety of binding activity.

Authors
Pisetsky, DS; Lerner, EA; Caster, S
MLA Citation
Pisetsky, DS, Lerner, EA, and Caster, S. "Idiotypic cross-reaction between MRL autoantibodies and a BALB/c myeloma." Mol Immunol 20.6 (June 1983): 615-621.
PMID
6877245
Source
pubmed
Published In
Molecular Immunology
Volume
20
Issue
6
Publish Date
1983
Start Page
615
End Page
621

Specificity and idiotype analysis of a monoclonal anti-DNA antibody

The binding properties and idiotypes of a monoclonal anti-DNA antibody were studied to investigate the control of specificity in this autoimmune response. This antibody, A9, was derived from the fusion of spleen cells of an MRL- lpr lpr mouse and the cell line X63-Ag8.653. In inhibition binding studies, A9 showed preference for single-stranded DNA and bound both ribo- and deoxyribohomopolymers. An antiserum prepared in a rabbit against A9 showed anti-idiotypic specificity after absorption with BALB c immunoglobulins and the BALB c myeloma MOPC 195. At least two distinct idiotypic specificities could be demonstrated by this serum, one apparently exclusive to A9 and one commonly expressed in sera of MRL- lpr lpr mice. Serum levels of the commonly expressed A9 idiotype were also elevated in sera of C3H mice expressing the lpr gene suggesting association with B-cell activation induced by this genetic mechanism. The demonstration of a commonly expressed A9 idiotype indicates that some autoantibody variable region determinants may be genetically controlled. © 1983.

Authors
Pisetsky, D
MLA Citation
Pisetsky, D. "Specificity and idiotype analysis of a monoclonal anti-DNA antibody." Clinical Immunology and Immunopathology 27.3 (1983): 348-356.
PMID
6191903
Source
scival
Published In
Clinical Immunology and Immunopathology
Volume
27
Issue
3
Publish Date
1983
Start Page
348
End Page
356

Idiotypic analysis of a monoclonal anti-Sm antibody.

Among murine models of autoimmunity, MRL mice are unique in their expression of antibodies to the nuclear antigen Sm. To assess genetic mechanisms in the control of this response, the idiotypes borne by a monoclonal anti-Sm antibody of MRL-Ipr/Ipr origin were investigated. Rabbit antisera were prepared against Y2, a hybridoma product with anti-Sm activity, and were rendered specific for idiotype by extensive absorption with normal globulins from BALB/c mice. In assays of idiotype by an inhibition ELISA, Y2 was shown to share idiotypes with Y12, another monoclonal anti-Sm derived from the same fusion as Y2; other monoclonal autoantibodies of MRL origin but different antigenic specificity failed to display idiotype activity in this assay. The presence of other anti-idiotypic specificities was revealed by absorption and elution of the anti-idiotype from an MRL globulin column; sera from both anti-Sm-positive and negative mice demonstrated these idiotypes. These results suggest that the predominant specificities detected by the anti-idiotype were unique to the monoclonal antibodies of the same animal, although there was also activity to idiotypes not related to anti-Sm binding molecules.

Authors
Pisetsky, DS; Lerner, EA
MLA Citation
Pisetsky, DS, and Lerner, EA. "Idiotypic analysis of a monoclonal anti-Sm antibody." J Immunol 129.4 (October 1982): 1489-1492.
PMID
6180012
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
129
Issue
4
Publish Date
1982
Start Page
1489
End Page
1492

The B-cell repertoire for autoantibodies: frequency of precursor cells for anti-DNA antibodies.

Authors
Pisetsky, DS; Caster, SA
MLA Citation
Pisetsky, DS, and Caster, SA. "The B-cell repertoire for autoantibodies: frequency of precursor cells for anti-DNA antibodies." Cell Immunol 72.2 (September 15, 1982): 294-305.
PMID
6983913
Source
pubmed
Published In
Cellular Immunology
Volume
72
Issue
2
Publish Date
1982
Start Page
294
End Page
305

Independent expression of autoantibodies in systemic lupus erythematosus.

Antibodies to components of the cell nucleus have been viewed as specific serological markers of systemic lupus erythematosus (SLE). To determine whether these autoantibodies exhibit common regulation of their expression, antibody levels have been quantitatively assessed in serial samples from patients producing at least 2 different antibody specificities. In a comparison of the peak antibody levels as a measure of immune responsiveness, the magnitude of the antiDNA response varied independently of either the antiSm or the antiRNP responses. Serial analysis with selected patients demonstrated that antiDNA levels fluctuated according to a pattern related to disease activity. In the same patients, however, antiSm and antiRNP antibodies showed little variation in level, with no consistent relationship to disease activity. Furthermore, following therapy, antiDNA levels fell while neither antiSm nor antiRNP levels showed significant alteration. These results suggest that in SLE, autoantibodies may arise from distinct immunoregulatory disturbances, each characterized by a unique relationship to disease activity and response to therapy.

Authors
McCarty, GA; Rice, JR; Bembe, ML; Pisetsky, DS
MLA Citation
McCarty, GA, Rice, JR, Bembe, ML, and Pisetsky, DS. "Independent expression of autoantibodies in systemic lupus erythematosus." J Rheumatol 9.5 (September 1982): 691-695.
PMID
6983576
Source
pubmed
Published In
The Journal of rheumatology
Volume
9
Issue
5
Publish Date
1982
Start Page
691
End Page
695

Ipr gene control of the anti-DNA antibody response.

The influence of the Ipr gene on the anti-DNA antibody response was investigated in MRL and B6 Ipr/Ipr inbred mice, MRL +/+ mice less than a yr of age produced low levels of anti-DNA antibody, whereas older animals of this strain demonstrated levels in some instances comparable to those of the more severely affected MRL Ipr/Ipr mice. This result indicates a tendency to autoreactivity in MRL mice independent of the Ipr gene. To determine whether other mice bearing the Ipr gene would also express autoantibodies, the anti-DNA antibody responses of B6 Ipr/Ipr mice were studied. This strain was development by matings to transfer the Ipr gene into another inbred background and allow evaluation of the action independent of other disturbances of the MRL mice. Mice of this strain produced antibodies to DNA, with female animals displaying significantly higher levels than males. This result demonstrates that the Ipr gene can stimulate autoantibody production in mice other than the MRL strain and does not require abnormalities unique to this background to potentiate autoreactivity.

Authors
Pisetsky, DS; Caster, SA; Roths, JB; Murphy, ED
MLA Citation
Pisetsky, DS, Caster, SA, Roths, JB, and Murphy, ED. "Ipr gene control of the anti-DNA antibody response." J Immunol 128.5 (May 1982): 2322-2325.
PMID
6801136
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
128
Issue
5
Publish Date
1982
Start Page
2322
End Page
2325

Binding specificity of a monoclonal anti-DNA antibody.

To investigate the interaction of DNA and anti-DNA antibodies in the immune complex disease of systemic lupus erythematosus, the fine specificity of binding of a monoclonal anti-DNA antibody was determined. This antibody, termed Cll, was derived from the fusion of spleen cells from an autoimmune MRL-lpr/lpr mouse with the myeloma cell line M45. In a solid-phase ELISA assay to measure anti-DNA activity, Cll showed preference for single stranded compared to double stranded DNA of animal origin. The Cll antibody also bound some deoxyribohomopolymers as well as ribohomopolymers, but failed to bind synthetic DNA duplexes. Defined size oligonucleotides with a size range of 2-(12-18) failed to inhibit the binding of Cll to single stranded DNA. This pattern of binding is consistent with the recognition of a unique structural determinant that can be represented by a variety of nucleic acids. The absence of antigenic activity among the oligonucleotides suggests that an extended polynucleotide structure is required for antibody binding, possibly because of a bivalent or 'monogamous' mode of interaction. The binding properties of Cll further suggest that its ability to participate in immune complex formation may be limited by the nature of the available DNA antigen.

Authors
Pisetsky, DS; Caster, SA
MLA Citation
Pisetsky, DS, and Caster, SA. "Binding specificity of a monoclonal anti-DNA antibody." Mol Immunol 19.5 (May 1982): 645-650.
PMID
7110140
Source
pubmed
Published In
Molecular Immunology
Volume
19
Issue
5
Publish Date
1982
Start Page
645
End Page
650

Lymphocytic lymphoma and systemic lupus erythematosus: their coexistence with antibody to the Sm antigen.

The serologic and clinical features of a 32-year-old women with coexistent systemic lupus erythematosus and lymphoma were studied. A cervical node biopsy specimen demonstrated nodular, poorly differentiated lymphocytic lymphoma; subsequent development of polyarthralgias prompted evaluation for connective-tissue disease, and a high titer of antinuclear antibody was found. Immunochemical studies proved that this antibody was directed to the Sm antigen, a serologic finding highly specific for systemic lupus erythematosus. This antibody has been maintained throughout the course of the patient's disease, characterized by persistent lymphadenopathy, pleuropericarditis, arthritis, hypocomplementemia, hyperglobulinemia, and a slight elevation of titers of anti-DNA antibody. The course of the lymphoproliferative disorder has been consistent with that of an indolent non-Hodgkin's lymphoma. In sera from other patients with lymphoproliferative disorders, anti-Sm antibody was not found. These findings emphasize the importance of establishing the antigenic specificity of antinuclear antibodies detected in the sera of patients with neoplasia.

Authors
McCarty, GA; Rice, JR; Fetter, BF; Pisetsky, DS
MLA Citation
McCarty, GA, Rice, JR, Fetter, BF, and Pisetsky, DS. "Lymphocytic lymphoma and systemic lupus erythematosus: their coexistence with antibody to the Sm antigen." Arch Pathol Lab Med 106.4 (April 1982): 196-199.
PMID
6175291
Source
pubmed
Published In
Archives of Pathology and Laboratory Medicine
Volume
106
Issue
4
Publish Date
1982
Start Page
196
End Page
199

Lpr gene control of the anti-DNA antibody response

The influence of the Lpr gene on the anti-DNA antibody response was investigated in MRL and B6 Lpr Lpr inbred mice. MRL +/+ mice less than a yr of age produced low levels of anti-DNA antibody, whereas older animals of this strain demonstrated levels in some instances comparable to those of the more severely affected MRL Lpr/Lpr mice. This result indicates a tendency to autoreactivity in MRL mice independent of the Lpr gene. To determine whether other mice bearing the Lpr gene would also express autoantibodies, the anti-DNA antibody responses of B6 Lpr/Lpr mice were studied. This strain was developed by matings to transfer the Lpr gene into another inbred background and allow evaluation of its action independent of other disturbances of the MRL mice. Mice of this strain produced antibodies to DNA, with female animals displaying significantly higher levels than males. This result demonstrates that the Lpr gene can stimulate autoantibody production in mice other than the MRL strain and does not require abnormalities unique to this background to potentiate autoreactivity.

Authors
Pisetsky, DS; Caster, SA; Roths, JB; Murphy, ED
MLA Citation
Pisetsky, DS, Caster, SA, Roths, JB, and Murphy, ED. "Lpr gene control of the anti-DNA antibody response." Journal of Immunology 128.5 (1982): 2322-2325.
Source
scival
Published In
Journal of Immunology
Volume
128
Issue
5
Publish Date
1982
Start Page
2322
End Page
2325

Ir genes of different high responder haplotypes for staphylococcal nuclease are not allelic.

The Ir gene controlling high responsiveness to staphylococcal nuclease in the H-2d haplotype has been mapped to the I-A subregion, in contrast to that in the H-2k and H-2a haplotypes, which maps in the I-B subregion. The nonallelic high responder genes also confer differences in fine specificity on the antibodies produced. This nonallelism of Ir genes for the same antigen in different haplotypes is consistent with Ir gene mechanisms involving associative recognition of antigen plus Ia by T lymphocytes. It further suggests that products of different subregions can perform the same function.

Authors
Berzofsky, JA; Pisetsky, DS; Killion, DJ; Hicks, G; Sachs, DH
MLA Citation
Berzofsky, JA, Pisetsky, DS, Killion, DJ, Hicks, G, and Sachs, DH. "Ir genes of different high responder haplotypes for staphylococcal nuclease are not allelic." J Immunol 127.6 (December 1981): 2453-2455.
PMID
6795275
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
127
Issue
6
Publish Date
1981
Start Page
2453
End Page
2455

The induction of tolerance and suppression in autoimmune MRL mice using hapten-modified self.

Disturbances in suppressor cell function have been considered important in the pathogenesis of systemic lupus erythematosus (SLE), a conclusion supported by studies with New Zealand mice. To determine whether other SLE mice display similar immunoregulatory defects, we investigated the susceptibility of autoimmune MRL mice to unresponsiveness induced by hapten-modified self (HMS). The response of splenocytes from MRL-lpr/lpr mice (lpr) was compared with those of sex- and age-matched, congenic MRL-+/+ mice (+/+), and H-2-identical (H-2k) CBA/J mice. Spleen cells (NSC) were cultured in vitro with hapten-modified syngeneic splenocytes (TNP-SC) and tested for responsiveness to TNP-LPS (for tolerance) or their ability to suppress the response of fresh cells. There was no difference in the susceptibility of lpr splenocytes from 3- and 10-mo-old mice to the induction of tolerance or suppression when compared with those from age-matched +/+ or CBA mice. To evaluate any quantitative defects in the responsiveness of lpr splenocytes to HMS, we modified the conditions under which suppressor activity was generated. Varying the ratio of NSC to TNP-SC from 10:1 to 2000:1, or changing the concentration of TNBS for haptenation from 10 mM to 0.5 mM per 10(8) spleen cells revealed no differences in the dose-response curves of lpr splenocytes for both tolerance and suppression when compared with those of the CBA. These results indicate that clinically affected MRL mice have intact suppressor cell activity in response to antigen-modified self and suggest a possible therapeutic role of this modality in inducing tolerance to self-antigens.

Authors
Santoro, TJ; Scott, DW; Pisetsky, DS
MLA Citation
Santoro, TJ, Scott, DW, and Pisetsky, DS. "The induction of tolerance and suppression in autoimmune MRL mice using hapten-modified self." J Immunol 127.2 (August 1981): 690-693.
PMID
7252156
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
127
Issue
2
Publish Date
1981
Start Page
690
End Page
693

A simple enzyme-linked immunosorbent assay for antibodies to native DNA.

A simple and highly sensitive enzyme-linked immunosorbent assay (ELISA) has been developed for the measurement of anti-native DNA (anti-nDNA) antibodies in sera from patients with systemic lupus erythematosus (SLE). A solid-phase support for the assay was provided by coating wells of polystyrene microtiter plates with native salmon sperm DNA at a concentration of 1 microgram/ml. For standard determinations, test sera at a dilution of 1 : 100 were incubated in the DNA-coated wells which were then assayed for bound immunoglobulin using an alkaline phosphatase conjugated rabbit anti-human IgG reagent. The colorimetric yield at 400 nm was used as a measure of the content of anti-nDNA antibodies. Proof that the bound antibodies detected in this assay were directed toward native DNA was derived primarily from observations that: (1) purified native DNA blocked the binding of antibody from SLE sera; and (2) there was a correlation between determinations by the ELISA assay and a filter binding assay specific for anti-nDNA antibodies. Assay of serial samples from the same patient showed a close correlation between determinations by the ELISA and filter binding assays, suggesting the utility of the ELISA method as part of the evaluation of disease activity over time. The advantages of the ELISA methodology-sensitivity, convenience, speed, independence of radioactivity, and quantitative determination of antibody as opposed to protein-should make this assay a useful tool in clinical and experimental studies on SLE as well as the routine assay of anti-nDNA antibodies.

Authors
Pisetsky, DS; Peters, DV
MLA Citation
Pisetsky, DS, and Peters, DV. "A simple enzyme-linked immunosorbent assay for antibodies to native DNA." J Immunol Methods 41.2 (1981): 187-200.
PMID
7021686
Source
pubmed
Published In
Journal of Immunological Methods
Volume
41
Issue
2
Publish Date
1981
Start Page
187
End Page
200

Mechanisms of autoantibody production in autoimmune MRL mice.

The quantitative expression of anti-DNA and anti-Sm antibodies has been investigated in autoimmune MRL-lpr/lpr and MRL-+/+ mice. Anti-Sm antibodies were detected in sera from 21/23 lpr/lpr and 10/16 +/+ mice, with individual animals showing striking variation in the time-course and magnitude of this autoantibody response. The peak antibody levels of the responding animals of each substrain did not differ significantly. For anti-DNA antibody, a different pattern of responsiveness was observed. Individual animals of each substrain produced very similar responses in terms of the magnitude and time-course of serum anti-DNA antibody. The differences in the peak levels of the two substrains were highly significant, with lpr/lpr mice demonstrating a much greater anti-DNA antibody response than +/+ mice. In lpr/lpr mice tested for both autoantibody systems, serum anti-DNA and anti-Sm antibodies showed distinct time-courses. These studies indicate that anti-DNA and anti-Sm antibodies are expressed independently in MRL mice, with the expression of anti-DNA, but not anti-Sm antibody markedly influenced by the presence of the 1pr gene. A fundamental difference in the mechanisms involved in the generation of anti-DNA and anti-Sm antibodies is suggested by the quantitative pattern of the two responses.

Authors
Pisetsky, DS; McCarty, GA; Peters, DV
MLA Citation
Pisetsky, DS, McCarty, GA, and Peters, DV. "Mechanisms of autoantibody production in autoimmune MRL mice." J Exp Med 152.5 (November 1, 1980): 1302-1310.
PMID
7430950
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
152
Issue
5
Publish Date
1980
Start Page
1302
End Page
1310

Genetic control of the immune response to staphylococcal nuclease. IX. Recombination between genes determining BALB/c antinuclease idiotypes and the heavy chain allotype locus.

The genetic linkage relationship of two antinuclease idiotypes produced by the BALB/c strain was investigated in the backcross (BALB/c x CB.20) X CB.20. These two idiotypes were detected by Lewis rat anti-idiotypic antisera prepared against affinity-purified A/J and SJL antinuclease antibodies, termed the A/J and SJL idiotypes, respectively. Both idiotypes were found to be linked to the IgCHa immunoglobulin heavy chain allotype locus. There was, however, a high frequency of recombination observed between both markers and the IgCHa locus, with eight of 83 backcross animals recombinant for the A/J idiotype and five of 83 recombinant for the SJL idiotype. All such recombinant animals were IgCHb/b homozygotes that had gained one or both idiotypes. These results are consistent with a genetic map of VHr region genes in the BALB/c strain in which genes determining the SJL idiotype are closer to the IgCHa allotype locus than are genes determining the A/J idiotype. This high frequency of recombination may indicate that the chromosome segment containing VH region genes is very large or that it has structural features that promote recombination.

Authors
Pisetsky, DS; Riordan, SE; Sachs, DH
MLA Citation
Pisetsky, DS, Riordan, SE, and Sachs, DH. "Genetic control of the immune response to staphylococcal nuclease. IX. Recombination between genes determining BALB/c antinuclease idiotypes and the heavy chain allotype locus." J Immunol 122.3 (March 1979): 842-846.
PMID
109505
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
122
Issue
3
Publish Date
1979
Start Page
842
End Page
846

Genetic control of the immune response to staphylococcal nuclease. VIII. Mapping of genes for antibodies to different antigenic regions of nuclease.

Antibodies to staphylococcal nuclease have been fractionated into two populations on the basis of their ability to bind to the cyanogen bromide cleavage product of nuclease comprising the C-terminal portion of the molecule from the 99th to the 149th amino acid. The two populations of antibodies, anti-nuclease (1-99)n and anti-nuclease (99-149)N, have been prepared from a variety of strains, and analyzed using anti-idiotypic antisera raised against whole anti-nuclease antibodies from strains A/J, SJL, BALB/c, and B10.A(2R). Anti-nuclease (1-99)n, antibodies had the same pattern of reactivity with the anti-idiotypic antisera as did unfractionated antibodies, whereas a different pattern was found for anti-nuclease (99-149)n preparations. On the basis of these studies, five anti-nuclease idiotypes, designated NASE markers, have been identified and defined on the basis of their antigenic specificity and strain distribution. With these additional markers, it has been possible to provide more detailed maps of variable (V) region genes in the strains BALB/c, CB.20, and the recombinant BAB.14. A recombinational event between V region genes during the development of the BAB.14 strain is suggested by the positioning of these NASE markers.

Authors
Pisetsky, DS; Sachs, DH
MLA Citation
Pisetsky, DS, and Sachs, DH. "Genetic control of the immune response to staphylococcal nuclease. VIII. Mapping of genes for antibodies to different antigenic regions of nuclease." J Exp Med 147.5 (May 1, 1978): 1517-1525.
PMID
77307
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
147
Issue
5
Publish Date
1978
Start Page
1517
End Page
1525

Genetic control of the immune response to staphylococcal nuclease. VII. Role of non-H2-linked genes in the control of the anti-nuclease antibody response.

The role of non-H-2-linked genes in the control of the antibody response to staphylococcal nuclease has been investigated. 3 wk after immunization with nuclease in complete Freund's adjuvant, strain A/J (H-2 a) mice produced significantly higher titers of antibody than strain B10.A (H-2(a)) mice, whereas mice of strains A.BY (H-2(b)) and B10 (H-2(b)) produced barely detectable titers. With hyperimmunization, A/J and A.BY mice reached the same peak levels for antibody titers, both severalfold higher than those reached by B10.A and B10 mice. Analysis of the specificity of antibodies by assessment of binding to two fragments of nuclease showed similarities between strains of the same H-2 haplotype. These results suggest that although H-2-1inked genes determined initial responsiveness at 3 wk and the relative proportions of antibodies directed toward different antigenic determinants on the nuclease molecule, non-H-2-linked genes determined the overall magnitude of the hyperimmuneresponse. Measurement of the affinity of the antibodies to the nuclease fragment (1-126) showed that strains B10 and B10.A produced antibodies with 7- to 10-fold higher affinity than comparable antibodies from strains A.BY and A/J. In a backcross of (B10.A x A/J) x B10.A, the level of antibody segregated independently of the Ig-1(e) C(H) allotype and the A/J anti-nuclease idiotypes. Thus, a gene(s) linked to neither H-2 nor heavy chain structural genes appears to control the aggregate response to antigenic determinants on the nuclease molecule independent of subspecificities of these antibodies or their idiotype.

Authors
Pisetsky, DS; Berzofsky, JA; Sachs, DH
MLA Citation
Pisetsky, DS, Berzofsky, JA, and Sachs, DH. "Genetic control of the immune response to staphylococcal nuclease. VII. Role of non-H2-linked genes in the control of the anti-nuclease antibody response." J Exp Med 147.2 (February 1, 1978): 396-408.
PMID
415108
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
147
Issue
2
Publish Date
1978
Start Page
396
End Page
408

Genetic control of the immune response to staphylococcal nuclease in mice.

Genetic control of the immune response to staphylococcal nuclease in mice is detectable at several levels. At least one H-2-linked Ir gene controls 1) the relative proportions of antibodies to different determinants on nuclease when whole nuclease is the immunogen; 2) the immunogenicity of isolated fragments of nuclease, corresponding to the same regions or determinants; and 3) the T-lymphocyte proliferative response to nuclease and to its fragments. It is concluded that a model in which Ir-gene control is determined by the recognition by T lymphocytes of a single "carrier" determinant for the whole molecule does not adequately explain this system. Evidence is presented for the existence of more than one such H-2-linked Ir gene in the T-cell proliferative response. In addition, a non-H-2-linked gene(s) is described which controls the overall level of antibodies to nuclease, i.e., the aggregate of all the antibodies of different subspecificities which have in common that they bind to some part of the nuclease molecule. Evidence is also presented that T lymphocytes, as well as the receptors involved in Ir-gene function (whether or not these are T-lymphocyte receptors), are less sensitive to conformational differences between native nuclease and its isolated fragments than are the antibodies ultimately made. This insensitivity to conformation may reflect the recognition of determinants which are shorter or more flexible in the native state than those recognized by antibodies.

Authors
Berzofsky, JA; Pisetsky, DS; Schwartz, RH; Schechter, AN; Sachs, DH
MLA Citation
Berzofsky, JA, Pisetsky, DS, Schwartz, RH, Schechter, AN, and Sachs, DH. "Genetic control of the immune response to staphylococcal nuclease in mice." Adv Exp Med Biol 98 (1978): 241-257.
PMID
102125
Source
pubmed
Published In
Advances in experimental medicine and biology
Volume
98
Publish Date
1978
Start Page
241
End Page
257

Genetic control of the immune response to staphylococcal nuclease

The nuclease serves as a model antigen in the study of the genetic control of the immune response. The immune response to the total antigen is based on the component reactions to a multiplicity of various determinants. Studies on T cell proliferation have shown that the Ir gene control promotes complex reciprocal actions of a series of component reactivities. It is presumed that genes that determine idiotypes of anti-nuclease antibodies in their development are independent of H-2 coupled Ir genes, but nevertheless possess a close coupling with the H-chain allotype locus. (D.O. Schmid - Munchen)

Authors
Sachs, DH; Berzofsky, JA; Pisetsky, DS; Schwartz, RH
MLA Citation
Sachs, DH, Berzofsky, JA, Pisetsky, DS, and Schwartz, RH. "Genetic control of the immune response to staphylococcal nuclease." Springer Seminars in Immunopathology 1.1 (1978): 51-83.
Source
scival
Published In
Springer Seminars in Immunopathology
Volume
1
Issue
1
Publish Date
1978
Start Page
51
End Page
83

The genetic control of the immune response to staphylococcal nuclease VI. Recombination between genes determining the A/J anti-nuclease idiotypes and the heavy chain allotype locus.

Rat antisera detecting binding site-specific idiotypic determinants on anti-nuclease antibodies from A/J mice have been used to define the A/J anti-nuclease idiotype and to investigate its genetic linkage as a variable region marker. Analysis of the segregation of the A/J idiotype in progeny of the backcross (B10.A X A/J) X B10.A showed linkage of the idiotype to the Ig-1e heavy chain allotype locus. There was, however, a very high apparent frequency of recombination, with 7 of 101 backcross animals having a recombinant phenotype. All of these putative recombinants were accounted for by Ig-1b/Ig-1b homozygotes which bore the A/J idiotype, and none by Ig-1b/Ig-1e heterozygotes lacking the idiotype. On progeny testing of these animals in another backcross to B10.A the recombinant trait bred true. If this idiotype is indeed a marker for variable region structural genes, then the germ line gene pool must be very large or there must be special genetic mechanism to account for the increased recombinational frequency observed.

Authors
Pisetsky, DS; Sachs, DH
MLA Citation
Pisetsky, DS, and Sachs, DH. "The genetic control of the immune response to staphylococcal nuclease VI. Recombination between genes determining the A/J anti-nuclease idiotypes and the heavy chain allotype locus." J Exp Med 146.6 (December 1, 1977): 1603-1612.
PMID
411875
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
146
Issue
6
Publish Date
1977
Start Page
1603
End Page
1612

Genetic control of the immune response to nuclease. V. Genetic linkage and strain distribution of anti-nuclease idiotypes.

Rat antisera raised against anti-nuclease antibodies from mouse strains A/J and SJL detect strain-specific idiotypic determinants related to the antigen-combining site. These antisera have been used to investigate the genetic linkage and strain distribution of the anti-nuclease idiotypes. Despite the existence of an H-2-linked immune response gene controlling the humoral response to nuclease, expression of the A/J anti-nuclease idiotype has been shown to be independent of genes in the H-2 region: the A/J idiotype was present in immune sera from strains A/J (H-2a) and A.BY (H-2b) but absent in sera from strains B10 (H-2b) and B10.A (H-2a). An analysis of the segregation of the A/J idiotype in offspring of the backcross (A/J x B10.A) x B10.A demonstrated linkage to the Ig-1e heavy chain allotype markers. In a small sample of backcross animals a very high apparent recombination frequency was observed, but further backcross analyses and progeny testing of putative recombinant animals will be required to substantiate this observation. Analysis of the A/J and SJL anti-nuclease idiotype markers in the BALB/c, CB.20, and BAB.14 strains indicate that these idiotypic markers may permit mapping of distinct variable region genes.

Authors
Fathman, CG; Pisetsky, DS; Sachs, DH
MLA Citation
Fathman, CG, Pisetsky, DS, and Sachs, DH. "Genetic control of the immune response to nuclease. V. Genetic linkage and strain distribution of anti-nuclease idiotypes." J Exp Med 145.3 (March 1, 1977): 569-577.
PMID
233902
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
145
Issue
3
Publish Date
1977
Start Page
569
End Page
577

The immune response to staphylococcal nuclease: a probe of cellular and humoral antigen-specific receptors.

Authors
Sachs, DH; Berzofsky, JA; Fathman, CG; Pisetsky, DS; Schechter, AN; Schwartz, RH
MLA Citation
Sachs, DH, Berzofsky, JA, Fathman, CG, Pisetsky, DS, Schechter, AN, and Schwartz, RH. "The immune response to staphylococcal nuclease: a probe of cellular and humoral antigen-specific receptors." Cold Spring Harb Symp Quant Biol 41 Pt 1 (1977): 295-306.
PMID
70312
Source
pubmed
Published In
Cold Spring Harbor Laboratory: Symposia on Quantitative Biology
Volume
41 Pt 1
Publish Date
1977
Start Page
295
End Page
306

The immune response to staphylococcal nuclease: a probe of cellular and humoral antigen specific receptors Origins of Lymphocyte Diversity

Authors
Sachs, DH; Berzofsky, JA; Fathman, CG; Pisetsky, DS; Schechter, AN; Schwartz, RH
MLA Citation
Sachs, DH, Berzofsky, JA, Fathman, CG, Pisetsky, DS, Schechter, AN, and Schwartz, RH. "The immune response to staphylococcal nuclease: a probe of cellular and humoral antigen specific receptors Origins of Lymphocyte Diversity." Cold Spring Harbor Symposia on Quantitative Biology 41.1 (1976): 295-306.
Source
scival
Published In
Cold Spring Harbor Laboratory: Symposia on Quantitative Biology
Volume
41
Issue
1
Publish Date
1976
Start Page
295
End Page
306

Role of ATP in DNA synthesis in Escherichia coli

Escherichia coli, rendered permeable by toluene treatment, is known to catalyze a limited amount of semi-conservative DNA synthesis; in addition to the four deoxynucleoside triphosphates, the reaction specifically requires ATP (or ADP). We show that this requirement was also evident in the conversion of Okazaki fragments to high molecular weight DNA in strains of E. coli carrying the pol A1 mutation. In strains containing DNA polymerase I, ATP was not required for this conversion. In addition, strains containing DNA polymerase I convert Okazaki fragments to a high molecular weight form more rapidly than strains deficient in this enzyme. The influence of nalidixic acid on this conversion process has been studied. In the presence of nalidixic acid, the 6 to 10 s Okazaki fragments are converted into a collection of molecules sedimenting at 30 to 40 s; there is essentially no breakdown of the fragments to an acid-soluble form under these conditions. When synthesis of DNA is inhibited by incubating temperaturesensitive mutants of the dnaB locus at the non-permissive temperature, however, there is a preferential degradation of the Okazaki fragments into an acidsoluble form. Prior incubation at the permissive temperature will prevent this degradation. © 1972 Academic Press Inc. (London) Limited.

Authors
Pisetsky, D; Berkower, I; Wickner, R; Hurwitz, J
MLA Citation
Pisetsky, D, Berkower, I, Wickner, R, and Hurwitz, J. "Role of ATP in DNA synthesis in Escherichia coli." Journal of Molecular Biology 71.3 (1972): 557-571.
PMID
4567466
Source
scival
Published In
Journal of Molecular Biology
Volume
71
Issue
3
Publish Date
1972
Start Page
557
End Page
571

Thermal-Turbidimetric studies of membranes from Acholeplasma laidlawii

Light scattering measurements were made of aqueous dispersions of Acholeplasma laidlawii membranes as well as lipids extracted from these membranes. As the temperature was increased the turbidity values changed over a temperature range that was dependent upon the fatty acid residues present in the membrane lipids. These changes were reversible and followed a pattern similar to the changes observed in calorimetric studies. The advantage of this procedure is that it permits thermal studies using relatively small amounts of membranes and also permits the study of some interactions between the membrane components and solutes in the aqueous medium. In this way an interaction was observed with Ca2+ occuring only when the membrane had changed to the fluid state. © 1972.

Authors
Abramson, MB; Pisetsky, D
MLA Citation
Abramson, MB, and Pisetsky, D. "Thermal-Turbidimetric studies of membranes from Acholeplasma laidlawii." BBA - Biomembranes 282.C (1972): 80-84.
PMID
5070088
Source
scival
Published In
BBA - Biomembranes
Volume
282
Issue
C
Publish Date
1972
Start Page
80
End Page
84
DOI
10.1016/0005-2736(72)90312-4

Are mycoplasma membrane proteins affected by variations in membrane fatty acid composition?

1. 1. The influence of the fatty acid composition of membrane lipids on the quantitative distribution of membrane protein of Mycoplasma laidlawii has been investigated. 2. 2. Cells were grown in media supplemented with either a saturated fatty acid (palmitic acid or stearic acid) or an unsaturated fatty acid (palmitoleic acid or oleic acid). Lipids isolated from membranes of these cells were markedly enriched in the supplemented fatty acid. 3. 3. The proteins of the different types of membranes were analyzed by sodium dodecyl sulfate-acrylamide-gel electrophoresis of membranes from cells grown in the presence of radioactively labelled amino acids. In general, protein composition and relative abundance were unaffected by fatty acid variation. One high molecular weight protein fraction was present in higher concentration in membranes rich in saturated fatty acids than in those rich in saturated fatty acids. 4. 4. The membranes rich in saturated fatty acids were also found to have a slightly higher density as determined by isopycnic centrifugation. © 1972.

Authors
Pisetsky, D; Terry, TM
MLA Citation
Pisetsky, D, and Terry, TM. "Are mycoplasma membrane proteins affected by variations in membrane fatty acid composition?." BBA - Biomembranes 274.1 (1972): 95-104.
PMID
5044067
Source
scival
Published In
BBA - Biomembranes
Volume
274
Issue
1
Publish Date
1972
Start Page
95
End Page
104

Microparticles as autoadjuvants in the pathogenesis of SLE.

Nucleic acids represent the main source of autoantigens in systemic lupus erythematosus (SLE). DNA and RNA can exit the cell during cell death and, in the extracellular space, can be immunostimulatory. Also extracellularly, DNA and RNA can be incorporated into microparticles (MPs)-small, membrane-bound vesicles released from dying cells by blebbing. We suggest that MPs display autoantigens, such as RNA and DNA, in a highly immunostimulatory manner, enabling them to function as autoadjuvants. In the bone marrow, nucleic-acid-containing MP autoadjuvants might induce B-cell tolerance, whereas in the periphery, they might stimulate mature B cells that have escaped central tolerance. Indeed, because MP autoadjuvants can trigger several receptors, they could effectively provide apoptotic or activating signals to B cells. We would therefore advance the idea that a model for SLE based on MP autoadjuvants can provide a new paradigm to elucidate the mechanisms by which DNA and RNA affect the immune system and critically influence B-cell fate.

Authors
Pisetsky, DS; Lipsky, PE
MLA Citation
Pisetsky, DS, and Lipsky, PE. "Microparticles as autoadjuvants in the pathogenesis of SLE." Nat Rev Rheumatol 6.6: 368-372. (Review)
PMID
20458331
Source
pubmed
Published In
Nature Reviews Rheumatology
Volume
6
Issue
6
Start Page
368
End Page
372
DOI
10.1038/nrrheum.2010.66

Are autoantibodies the targets of B-cell-directed therapy?

B-cell-directed therapy-the use of agents that eliminate B cells or block cytokines important for B-cell function-is emerging as a promising approach to the treatment of rheumatic disease. Target diseases, including systemic lupus erythematosus (SLE), display diverse patterns of autoantibody production and aberrant activation of B cells. Despite the success of this general approach, the mechanisms by which B-cell-directed therapy ameliorates disease, and the role of autoantibodies as biomarkers of clinical response remain unclear. Importantly, although B-cell-directed therapy can reduce the production of some autoantibodies, the effects can be variable and heterogeneous, probably reflecting the critical (but ill-defined) roles of different B-cell and plasma cell populations in autoantibody production. Future studies during clinical trials of these agents are needed to define which B-cell and autoantibody populations are affected (or ought to be), and to discover informative biomarkers of clinical response that can be used to advance this therapeutic approach.

Authors
Pisetsky, DS; Grammer, AC; Ning, TC; Lipsky, PE
MLA Citation
Pisetsky, DS, Grammer, AC, Ning, TC, and Lipsky, PE. "Are autoantibodies the targets of B-cell-directed therapy? (Published online)." Nat Rev Rheumatol 7.9: 551-556. (Review)
PMID
21808289
Source
pubmed
Published In
Nature Reviews Rheumatology
Volume
7
Issue
9
Start Page
551
End Page
556
DOI
10.1038/nrrheum.2011.108
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