You are here

Kelsoe, Garnett H.

Overview:

1. Lymphocyte development and antigen-driven diversification of immunoglobulin and T cell antigen receptor genes.
2. The germinal center reaction and mechanisms for clonal selection and self - tolerance. The origins of autoimmunity.
3. Interaction of innate- and adaptive immunity and the role of inflammation in lymphoid organogenesis.
4. The role of secondary V(D)J gene rearrangment in lymphocyte development and malignancies.
5. Mathematical modeling of immune responses, DNA motifs, collaborations in bioinformatics.
6. Humoral immunity to influenza and HIV-1.

Positions:

James B. Duke Professor of Immunology

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:

D.Sc. 1979

D.Sc. — Harvard University

Assistant Professor, Microbiology

University of Texas, Medical Branch at Galveston

Associate Professor, Microbiology

University of Texas, Medical Branch at Galveston

Associate Professor, Microbiology & Immunology

University of Maryland School of Medicine

Professor, Microbiology And Immunology

University of Maryland School of Medicine

Grants:

Basic Immunology Training Program

Administered By
Immunology
AwardedBy
National Institutes of Health
Role
Mentor
Start Date
July 01, 2014
End Date
June 30, 2019

Testing the Nearest Neighbor Approach to Active Vaccination for HIV-1

Administered By
Immunology
AwardedBy
University of Washington
Role
Principal Investigator
Start Date
November 07, 2014
End Date
September 30, 2017

Training Program in Inflammatory and Immunological Diseases

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

Structure-function analysis of infection- and vaccine-induced B-cell repertoires

Administered By
Immunology
AwardedBy
Boston Children's Hospital
Role
Principal Investigator
Start Date
August 01, 2014
End Date
July 31, 2017

Serial Block Face Scanning Electron Microscope

Administered By
Pathology
AwardedBy
National Institutes of Health
Role
Major User
Start Date
June 01, 2016
End Date
May 31, 2017

Immunity to novel T/F SHIVs: variability in the co-evolution of virus and host immunity

Administered By
Immunology
AwardedBy
Quality Biological, Inc.
Role
Principal Investigator
Start Date
December 01, 2016
End Date
April 30, 2017

Modeling affinity maturation at molecular resolution

Administered By
Immunology
AwardedBy
Boston University
Role
Principal Investigator
Start Date
April 15, 2015
End Date
March 31, 2017

HSV-2 Glycoprotein D Deletion Virus Elicits FcR Dependent

Administered By
Immunology
AwardedBy
Albert Einstein College of Medicine
Role
Principal Investigator
Start Date
March 01, 2016
End Date
February 28, 2017

Culturing and isolating human B-cells using Athena technology

Administered By
Immunology
AwardedBy
Immune Tolerance Network
Role
Principal Investigator
Start Date
May 01, 2015
End Date
January 31, 2016

Multiscale Systems Immunology

Administered By
Immunology
AwardedBy
National Institutes of Health
Role
Co Investigator
Start Date
September 30, 2010
End Date
September 29, 2015

Eliciting B cells to produce anti-HIV gp41 MPER-specific neutralizing antibodies

Administered By
Immunology
AwardedBy
Dana Farber Cancer Institute
Role
Principal Investigator
Start Date
September 01, 2010
End Date
August 31, 2015

BAFF Pathology: Novel Therapeutic Targets in Chronic Graft versus Host Disease

Administered By
Medicine, Cellular Therapy
AwardedBy
National Institutes of Health
Role
Mentor
Start Date
July 12, 2011
End Date
June 30, 2015

Microsimulation of anthracis-immune system interaction

Administered By
Biostatistics & Bioinformatics
AwardedBy
National Institutes of Health
Role
Investigator
Start Date
June 15, 2004
End Date
May 30, 2007
Show More

Publications:

Immunodominance of Antibody Recognition of the HIV Envelope V2 Region in Ig-Humanized Mice.

In the RV144 gp120 HIV vaccine trial, decreased transmission risk was correlated with Abs that reacted with a linear epitope at a lysine residue at position 169 (K169) in the HIV-1 envelope (Env) V2 region. The K169 V2 response was restricted to Abs bearing Vλ rearrangements that expressed aspartic acid/glutamic acid in CDR L2. The AE.A244 gp120 in AIDSVAX B/E also bound to the unmutated ancestor of a V2-glycan broadly neutralizing Ab, but this Ab type was not induced in the RV144 trial. In this study, we sought to determine whether immunodominance of the V2 linear epitope could be overcome in the absence of human Vλ rearrangements. We immunized IgH- and Igκ-humanized mice with the AE.A244 gp120 Env. In these mice, the V2 Ab response was focused on a linear epitope that did not include K169. V2 Abs were isolated that used the same human VH gene segment as an RV144 V2 Ab but paired with a mouse λ L chain. Structural characterization of one of these V2 Abs revealed how the linear V2 epitope could be engaged, despite the lack of aspartic acid/glutamic acid encoded in the mouse repertoire. Thus, despite the absence of the human Vλ locus in these humanized mice, the dominance of Vλ pairing with human VH for HIV-1 Env V2 recognition resulted in human VH pairing with mouse λ L chains instead of allowing otherwise subdominant V2-glycan broadly neutralizing Abs to develop.

Authors
Wiehe, K; Nicely, NI; Lockwood, B; Kuraoka, M; Anasti, K; Arora, S; Bowman, CM; Stolarchuk, C; Parks, R; Lloyd, KE; Xia, S-M; Duffy, R; Shen, X; Kyratsous, CA; Macdonald, LE; Murphy, AJ; Scearce, RM; Moody, MA; Alam, SM; Verkoczy, L; Tomaras, GD; Kelsoe, G; Haynes, BF
MLA Citation
Wiehe, K, Nicely, NI, Lockwood, B, Kuraoka, M, Anasti, K, Arora, S, Bowman, CM, Stolarchuk, C, Parks, R, Lloyd, KE, Xia, S-M, Duffy, R, Shen, X, Kyratsous, CA, Macdonald, LE, Murphy, AJ, Scearce, RM, Moody, MA, Alam, SM, Verkoczy, L, Tomaras, GD, Kelsoe, G, and Haynes, BF. "Immunodominance of Antibody Recognition of the HIV Envelope V2 Region in Ig-Humanized Mice." Journal of immunology (Baltimore, Md. : 1950) 198.3 (February 2017): 1047-1055.
PMID
28011932
Source
epmc
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
198
Issue
3
Publish Date
2017
Start Page
1047
End Page
1055
DOI
10.4049/jimmunol.1601640

BCR and Endosomal TLR Signals Synergize to Increase AID Expression and Establish Central B Cell Tolerance.

Activation-induced cytidine deaminase (AID) is required to purge autoreactive immature and transitional-1 (immature/T1) B cells at the first tolerance checkpoint, but how AID selectively removes self-reactive B cells is unclear. We now show that B cell antigen receptor (BCR) and endosomal Toll-like receptor (TLR) signals synergize to elicit high levels of AID expression in immature/T1 B cells. This synergy is restricted to ligands for endocytic TLR and requires phospholipase-D activation, endosomal acidification, and MyD88. The first checkpoint is significantly impaired in AID- or MyD88-deficient mice and in mice doubly heterozygous for AID and MyD88, suggesting interaction of these factors in central B cell tolerance. Moreover, administration of chloroquine, an inhibitor of endosomal acidification, results in a failure to remove autoreactive immature/T1 B cells in mice. We propose that a BCR/TLR pathway coordinately establishes central tolerance by hyper-activating AID in immature/T1 B cells that bind ligands for endosomal TLRs.

Authors
Kuraoka, M; Snowden, PB; Nojima, T; Verkoczy, L; Haynes, BF; Kitamura, D; Kelsoe, G
MLA Citation
Kuraoka, M, Snowden, PB, Nojima, T, Verkoczy, L, Haynes, BF, Kitamura, D, and Kelsoe, G. "BCR and Endosomal TLR Signals Synergize to Increase AID Expression and Establish Central B Cell Tolerance." Cell reports 18.7 (February 2017): 1627-1635.
PMID
28199836
Source
epmc
Published In
Cell Reports
Volume
18
Issue
7
Publish Date
2017
Start Page
1627
End Page
1635
DOI
10.1016/j.celrep.2017.01.050

Host controls of HIV broadly neutralizing antibody development.

Induction of broadly neutralizing antibodies (bNAbs) is a major goal of HIV vaccine development. BNAbs are made during HIV infection by a subset of individuals but currently cannot be induced in the setting of vaccination. Considerable progress has been made recently in understanding host immunologic controls of bNAb induction and maturation in the setting of HIV infection, and point to key roles for both central and peripheral immunologic tolerance mechanisms in limiting bnAb development. Immune tolerance checkpoint inhibition has been transformative in promotion of anti-tumor CD8 T-cell responses in the treatment of certain malignancies. Here, we review the evidence for host controls of bNAb responses, and discuss strategies for the transient modulation of immune responses with vaccines toward the goal of enhancing germinal center B-cell responses to favor bNAb B-cell lineages and to foster their maturation to full neutralization potency.

Authors
Kelsoe, G; Haynes, BF
MLA Citation
Kelsoe, G, and Haynes, BF. "Host controls of HIV broadly neutralizing antibody development." Immunological reviews 275.1 (January 2017): 79-88. (Review)
PMID
28133807
Source
epmc
Published In
Immunological Reviews
Volume
275
Issue
1
Publish Date
2017
Start Page
79
End Page
88
DOI
10.1111/imr.12508

HIV-1 Envelope Mimicry of Host Enzyme Kynureninase Does Not Disrupt Tryptophan Metabolism.

The HIV-1 envelope protein (Env) has evolved to subvert the host immune system, hindering viral control by the host. The tryptophan metabolic enzyme kynureninase (KYNU) is mimicked by a portion of the HIV Env gp41 membrane proximal region (MPER) and is cross-reactive with the HIV broadly neutralizing Ab (bnAb) 2F5. Molecular mimicry of host proteins by pathogens can lead to autoimmune disease. In this article, we demonstrate that neither the 2F5 bnAb nor HIV MPER-KYNU cross-reactive Abs elicited by immunization with an MPER peptide-liposome vaccine in 2F5 bnAb VHDJH and VLJL knock-in mice and rhesus macaques modified KYNU activity or disrupted tissue tryptophan metabolism. Thus, molecular mimicry by HIV-1 Env that promotes the evasion of host anti-HIV-1 Ab responses can be directed toward nonfunctional host protein epitopes that do not impair host protein function. Therefore, the 2F5 HIV Env gp41 region is a key and safe target for HIV-1 vaccine development.

Authors
Bradley, T; Yang, G; Ilkayeva, O; Holl, TM; Zhang, R; Zhang, J; Santra, S; Fox, CB; Reed, SG; Parks, R; Bowman, CM; Bouton-Verville, H; Sutherland, LL; Scearce, RM; Vandergrift, N; Kepler, TB; Moody, MA; Liao, H-X; Alam, SM; McLendon, R; Everitt, JI; Newgard, CB; Verkoczy, L; Kelsoe, G; Haynes, BF
MLA Citation
Bradley, T, Yang, G, Ilkayeva, O, Holl, TM, Zhang, R, Zhang, J, Santra, S, Fox, CB, Reed, SG, Parks, R, Bowman, CM, Bouton-Verville, H, Sutherland, LL, Scearce, RM, Vandergrift, N, Kepler, TB, Moody, MA, Liao, H-X, Alam, SM, McLendon, R, Everitt, JI, Newgard, CB, Verkoczy, L, Kelsoe, G, and Haynes, BF. "HIV-1 Envelope Mimicry of Host Enzyme Kynureninase Does Not Disrupt Tryptophan Metabolism." Journal of immunology (Baltimore, Md. : 1950) 197.12 (December 2016): 4663-4673.
PMID
27849170
Source
epmc
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
197
Issue
12
Publish Date
2016
Start Page
4663
End Page
4673

Efficient culture of human naive and memory B cells for use as APCs

© Copyright 2016 by The American Association of Immunologists, Inc. All rights reserved.The ability to culture and expand B cells in vitro has become a useful tool for studying human immunity. A limitation of current methods for human B cell culture is the capacity to support mature B cell proliferation.We developed a culture method to support the efficient activation and proliferation of naive and memory human B cells. This culture supports extensive B cell proliferation, with ∼103-fold increases following 8 d in culture and 106-fold increases when cultures are split and cultured for 8 more days. In culture, a significant fraction of naive B cells undergo isotype switching and differentiate into plasmacytes. Culture-derived (CD) B cells are readily cryopreserved and, when recovered, retain their ability to proliferate and differentiate. Significantly, proliferating CD B cells express high levels of MHC class II, CD80, and CD86. CD B cells act as APCs and present alloantigens and microbial Ags to T cells.We are able to activate and expand Ag-specific memory B cells; these cultured cells are highly effective in presenting Ag to T cells. We characterized the TCR repertoire of rare Ag-specific CD4+ T cells that proliferated in response to tetanus toxoid (TT) presented by autologous CD B cells. TCR Vb usage by TT-activated CD4+ T cells differs from resting and unspecifically activated CD4+ T cells. Moreover, we found that TT-specific TCR Vb usage by CD4+ T cells was substantially different between donors. This culture method provides a platform for studying the BCR and TCR repertoires within a single individual.

Authors
Su, KY; Watanabe, A; Yeh, CH; Kelsoe, G; Kuraoka, M
MLA Citation
Su, KY, Watanabe, A, Yeh, CH, Kelsoe, G, and Kuraoka, M. "Efficient culture of human naive and memory B cells for use as APCs." Journal of Immunology 197.10 (November 15, 2016): 4163-4176.
Source
scopus
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
197
Issue
10
Publish Date
2016
Start Page
4163
End Page
4176
DOI
10.4049/jimmunol.1502193

Identification of a CD4-Binding-Site Antibody to HIV that Evolved Near-Pan Neutralization Breadth.

Detailed studies of the broadly neutralizing antibodies (bNAbs) that underlie the best available examples of the humoral immune response to HIV are providing important information for the development of therapies and prophylaxis for HIV-1 infection. Here, we report a CD4-binding site (CD4bs) antibody, named N6, that potently neutralized 98% of HIV-1 isolates, including 16 of 20 that were resistant to other members of its class. N6 evolved a mode of recognition such that its binding was not impacted by the loss of individual contacts across the immunoglobulin heavy chain. In addition, structural analysis revealed that the orientation of N6 permitted it to avoid steric clashes with glycans, which is a common mechanism of resistance. Thus, an HIV-1-specific bNAb can achieve potent, near-pan neutralization of HIV-1, making it an attractive candidate for use in therapy and prophylaxis.

Authors
Huang, J; Kang, BH; Ishida, E; Zhou, T; Griesman, T; Sheng, Z; Wu, F; Doria-Rose, NA; Zhang, B; McKee, K; O'Dell, S; Chuang, G-Y; Druz, A; Georgiev, IS; Schramm, CA; Zheng, A; Joyce, MG; Asokan, M; Ransier, A; Darko, S; Migueles, SA; Bailer, RT; Louder, MK; Alam, SM; Parks, R; Kelsoe, G; Von Holle, T; Haynes, BF; Douek, DC; Hirsch, V; Seaman, MS; Shapiro, L; Mascola, JR; Kwong, PD; Connors, M
MLA Citation
Huang, J, Kang, BH, Ishida, E, Zhou, T, Griesman, T, Sheng, Z, Wu, F, Doria-Rose, NA, Zhang, B, McKee, K, O'Dell, S, Chuang, G-Y, Druz, A, Georgiev, IS, Schramm, CA, Zheng, A, Joyce, MG, Asokan, M, Ransier, A, Darko, S, Migueles, SA, Bailer, RT, Louder, MK, Alam, SM, Parks, R, Kelsoe, G, Von Holle, T, Haynes, BF, Douek, DC, Hirsch, V, Seaman, MS, Shapiro, L, Mascola, JR, Kwong, PD, and Connors, M. "Identification of a CD4-Binding-Site Antibody to HIV that Evolved Near-Pan Neutralization Breadth." Immunity 45.5 (November 2016): 1108-1121.
PMID
27851912
Source
epmc
Published In
Immunity
Volume
45
Issue
5
Publish Date
2016
Start Page
1108
End Page
1121
DOI
10.1016/j.immuni.2016.10.027

Complex Antigens Drive Permissive Clonal Selection in Germinal Centers.

Germinal center (GC) B cells evolve toward increased affinity by a Darwinian process that has been studied primarily in genetically restricted, hapten-specific responses. We explored the population dynamics of genetically diverse GC responses to two complex antigens-Bacillus anthracis protective antigen and influenza hemagglutinin-in which B cells competed both intra- and interclonally for distinct epitopes. Preferred VH rearrangements among antigen-binding, naive B cells were similarly abundant in early GCs but, unlike responses to haptens, clonal diversity increased in GC B cells as early "winners" were replaced by rarer, high-affinity clones. Despite affinity maturation, inter- and intraclonal avidities varied greatly, and half of GC B cells did not bind the immunogen but nonetheless exhibited biased VH use, V(D)J mutation, and clonal expansion comparable to antigen-binding cells. GC reactions to complex antigens permit a range of specificities and affinities, with potential advantages for broad protection.

Authors
Kuraoka, M; Schmidt, AG; Nojima, T; Feng, F; Watanabe, A; Kitamura, D; Harrison, SC; Kepler, TB; Kelsoe, G
MLA Citation
Kuraoka, M, Schmidt, AG, Nojima, T, Feng, F, Watanabe, A, Kitamura, D, Harrison, SC, Kepler, TB, and Kelsoe, G. "Complex Antigens Drive Permissive Clonal Selection in Germinal Centers." Immunity 44.3 (March 2, 2016): 542-552.
PMID
26948373
Source
epmc
Published In
Immunity
Volume
44
Issue
3
Publish Date
2016
Start Page
542
End Page
552
DOI
10.1016/j.immuni.2016.02.010

Heavy-chain receptor editing unbound.

Authors
Kelsoe, G
MLA Citation
Kelsoe, G. "Heavy-chain receptor editing unbound." Proceedings of the National Academy of Sciences of the United States of America 112.8 (February 17, 2015): 2297-2298.
PMID
25691748
Source
epmc
Published In
Proceedings of the National Academy of Sciences of USA
Volume
112
Issue
8
Publish Date
2015
Start Page
2297
End Page
2298
DOI
10.1073/pnas.1501480112

Natural IgM Is produced by CD5- Plasma cells that occupy a distinct survival niche in bone marrow

Copyright © 2014 by The American Association of Immunologists, Inc.Natural IgMis constitutively present in the serum, where it aids in the early control of viral and bacterial expansions. Natural IgMalso plays a significant role in the prevention of autoimmune disease by promoting the clearance of cellular debris. Nevertheless, the origins of natural IgMhave not been precisely defined. Previous studies focused on the role of CD5+ B1 cells in the production of natural IgM, but we show in this article that a discrete population of CD52 IgMplasmablasts and plasma cells in the bone marrow (BM) produces the majority of serum IgM in resting mice. These Ab-secreting cells (ASC) originate from peritoneal cavity-resident cells, because transfer of peritoneal cells completely restores serum IgMand the specific compartment of BMASC in Rag1-deficient mice.We show that BM natural IgM ASC arise from a fetal-lineage progenitor that is neither B1a nor B1b, and that this IgM ASC compartment contains a substantial fraction of long-lived plasma cells that do not occupy the IgG plasma cell survival niche in the BM; instead, they are supported by IL-5. In summary, we identified the primary source of natural IgMand showed that these ASC are maintained long-term in a unique survival niche within the BM.

Authors
Reynolds, AE; Kuraoka, M; Kelsoe, G
MLA Citation
Reynolds, AE, Kuraoka, M, and Kelsoe, G. "Natural IgM Is produced by CD5- Plasma cells that occupy a distinct survival niche in bone marrow." Journal of Immunology 194.1 (January 1, 2015): 231-242.
Source
scopus
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
194
Issue
1
Publish Date
2015
Start Page
231
End Page
242
DOI
10.4049/jimmunol.1401203

Polyreactivity and autoreactivity among HIV-1 antibodies.

It is generally acknowledged that human broadly neutralizing antibodies (bNAbs) capable of neutralizing multiple HIV-1 clades are often polyreactive or autoreactive. Whereas polyreactivity or autoreactivity has been proposed to be crucial for neutralization breadth, no systematic, quantitative study of self-reactivity among nonneutralizing HIV-1 Abs (nNAbs) has been performed to determine whether poly- or autoreactivity in bNAbs is a consequence of chronic antigen (Ag) exposure and/or inflammation or a fundamental property of neutralization. Here, we use protein microarrays to assess binding to >9,400 human proteins and find that as a class, bNAbs are significantly more poly- and autoreactive than nNAbs. The poly- and autoreactive property is therefore not due to the infection milieu but rather is associated with neutralization. Our observations are consistent with a role of heteroligation for HIV-1 neutralization and/or structural mimicry of host Ags by conserved HIV-1 neutralization sites. Although bNAbs are more mutated than nNAbs as a group, V(D)J mutation per se does not correlate with poly- and autoreactivity. Infrequent poly- or autoreactivity among nNAbs implies that their dominance in humoral responses is due to the absence of negative control by immune regulation. Interestingly, four of nine bNAbs specific for the HIV-1 CD4 binding site (CD4bs) (VRC01, VRC02, CH106, and CH103) bind human ubiquitin ligase E3A (UBE3A), and UBE3A protein competitively inhibits gp120 binding to the VRC01 bNAb. Among these four bNAbs, avidity for UBE3A was correlated with neutralization breadth. Identification of UBE3A as a self-antigen recognized by CD4bs bNAbs offers a mechanism for the rarity of this bNAb class.Eliciting bNAbs is key for HIV-1 vaccines; most Abs elicited by HIV-1 infection or immunization, however, are strain specific or nonneutralizing, and unsuited for protection. Here, we compare the specificities of bNAbs and nNAbs to demonstrate that bNAbs are significantly more poly- and autoreactive than nNAbs. The strong association of poly- and autoreactivity with bNAbs, but not nNAbs from infected patients, indicates that the infection milieu, chronic inflammation and Ag exposure, CD4 T-cell depletion, etc., alone does not cause poly- and autoreactivity. Instead, these properties are fundamentally linked to neutralization breadth, either by the requirement for heteroligation or the consequence of host mimicry by HIV-1. Indeed, we show that human UBE3A shares an epitope(s) with HIV-1 envelope recognized by four CD4bs bNAbs. The poly- and autoreactivity of bNAbs surely contribute to the rarity of membrane-proximal external region (MPER) and CD4bs bNAbs and identify a roadblock that must be overcome to induce protective vaccines.

Authors
Liu, M; Yang, G; Wiehe, K; Nicely, NI; Vandergrift, NA; Rountree, W; Bonsignori, M; Alam, SM; Gao, J; Haynes, BF; Kelsoe, G
MLA Citation
Liu, M, Yang, G, Wiehe, K, Nicely, NI, Vandergrift, NA, Rountree, W, Bonsignori, M, Alam, SM, Gao, J, Haynes, BF, and Kelsoe, G. "Polyreactivity and autoreactivity among HIV-1 antibodies." Journal of virology 89.1 (January 2015): 784-798.
PMID
25355869
Source
epmc
Published In
Journal of virology
Volume
89
Issue
1
Publish Date
2015
Start Page
784
End Page
798
DOI
10.1128/jvi.02378-14

TSC1 Promotes B Cell Maturation but Is Dispensable for Germinal Center Formation.

Accumulating evidence indicates that the tuberous sclerosis complex 1 (TSC1), a tumor suppressor that acts by inhibiting mTOR signaling, plays an important role in the immune system. We report here that TSC1 differentially regulates mTOR complex 1 (mTORC1) and mTORC2/Akt signaling in B cells. TSC1 deficiency results in the accumulation of transitional-1 (T1) B cells and progressive losses of B cells as they mature beyond the T1 stage. Moreover, TSC1KO mice exhibit a mild defect in the serum antibody responses or rate of Ig class-switch recombination after immunization with a T-cell-dependent antigen. In contrast to a previous report, we demonstrate that both constitutive Peyer's patch germinal centers (GCs) and immunization-induced splenic GCs are unimpaired in TSC1-deficient (TSC1KO) mice and that the ratio of GC B cells to total B cells is comparable in WT and TSC1KO mice. Together, our data demonstrate that TSC1 plays important roles for B cell development, but it is dispensable for GC formation and serum antibody responses.

Authors
Ci, X; Kuraoka, M; Wang, H; Carico, Z; Hopper, K; Shin, J; Deng, X; Qiu, Y; Unniraman, S; Kelsoe, G; Zhong, X-P
MLA Citation
Ci, X, Kuraoka, M, Wang, H, Carico, Z, Hopper, K, Shin, J, Deng, X, Qiu, Y, Unniraman, S, Kelsoe, G, and Zhong, X-P. "TSC1 Promotes B Cell Maturation but Is Dispensable for Germinal Center Formation." PloS one 10.5 (January 2015): e0127527-.
Website
http://hdl.handle.net/10161/10894
PMID
26000908
Source
epmc
Published In
PloS one
Volume
10
Issue
5
Publish Date
2015
Start Page
e0127527
DOI
10.1371/journal.pone.0127527

The Cellular and Molecular Biology of HIV-1 Broadly Neutralizing Antibodies

© 2015 Elsevier Ltd All rights reserved.Induction of neutralizing antibodies capable of protecting against human immunodeficiency virus 1 (HIV-1) infection is a key goal of HIV-1 vaccine development. The target of neutralizing antibodies is the HIV-1 envelope (Env) protein on the virion surface. Although HIV-1 is extraordinarily diverse, some antibodies can recognize highly conserved sites on Env and neutralize diverse viral strains. Such broadly neutralizing antibodies (bnAbs) arise in ~20% of infected individuals, usually several years after infection. Antibody responses to these conserved epitopes are disfavored and subdominant, and to date, no vaccines have induced bnAbs. Recent data have demonstrated that bnAbs often exhibit unusual characteristics, including long heavy chain complementarity determinant region 3, high levels of somatic mutation, and recognition of self-antigens, or autoreactivity-traits that render them prone to elimination by clonal deletion and limit their abundance in the B cell repertoire. These challenges have necessitated an in depth understanding of the maturation pathways of bnAbs and their immunoregulatory controls so that novel strategies can be designed to induce them by vaccination.

Authors
Haynes, BF; Saunders, KO; Kelsoe, G; Mascola, JR; Nabel, GJ
MLA Citation
Haynes, BF, Saunders, KO, Kelsoe, G, Mascola, JR, and Nabel, GJ. "The Cellular and Molecular Biology of HIV-1 Broadly Neutralizing Antibodies." Molecular Biology of B Cells: Second Edition. December 15, 2014. 441-461.
Source
scopus
Publish Date
2014
Start Page
441
End Page
461
DOI
10.1016/B978-0-12-397933-9.00024-2

Antibody light-chain-restricted recognition of the site of immune pressure in the RV144 HIV-1 vaccine trial is phylogenetically conserved.

In HIV-1, the ability to mount antibody responses to conserved, neutralizing epitopes is critical for protection. Here we have studied the light chain usage of human and rhesus macaque antibodies targeted to a dominant region of the HIV-1 envelope second variable (V2) region involving lysine (K) 169, the site of immune pressure in the RV144 vaccine efficacy trial. We found that humans and rhesus macaques used orthologous lambda variable gene segments encoding a glutamic acid-aspartic acid (ED) motif for K169 recognition. Structure determination of an unmutated ancestor antibody demonstrated that the V2 binding site was preconfigured for ED motif-mediated recognition prior to maturation. Thus, light chain usage for recognition of the site of immune pressure in the RV144 trial is highly conserved across species. These data indicate that the HIV-1 K169-recognizing ED motif has persisted over the diversification between rhesus macaques and humans, suggesting an evolutionary advantage of this antibody recognition mode.

Authors
Wiehe, K; Easterhoff, D; Luo, K; Nicely, NI; Bradley, T; Jaeger, FH; Dennison, SM; Zhang, R; Lloyd, KE; Stolarchuk, C; Parks, R; Sutherland, LL; Scearce, RM; Morris, L; Kaewkungwal, J; Nitayaphan, S; Pitisuttithum, P; Rerks-Ngarm, S; Sinangil, F; Phogat, S; Michael, NL; Kim, JH; Kelsoe, G; Montefiori, DC; Tomaras, GD; Bonsignori, M; Santra, S; Kepler, TB; Alam, SM; Moody, MA; Liao, H-X; Haynes, BF
MLA Citation
Wiehe, K, Easterhoff, D, Luo, K, Nicely, NI, Bradley, T, Jaeger, FH, Dennison, SM, Zhang, R, Lloyd, KE, Stolarchuk, C, Parks, R, Sutherland, LL, Scearce, RM, Morris, L, Kaewkungwal, J, Nitayaphan, S, Pitisuttithum, P, Rerks-Ngarm, S, Sinangil, F, Phogat, S, Michael, NL, Kim, JH, Kelsoe, G, Montefiori, DC, Tomaras, GD, Bonsignori, M, Santra, S, Kepler, TB, Alam, SM, Moody, MA, Liao, H-X, and Haynes, BF. "Antibody light-chain-restricted recognition of the site of immune pressure in the RV144 HIV-1 vaccine trial is phylogenetically conserved." Immunity 41.6 (December 2014): 909-918.
PMID
25526306
Source
epmc
Published In
Immunity
Volume
41
Issue
6
Publish Date
2014
Start Page
909
End Page
918
DOI
10.1016/j.immuni.2014.11.014

Curiouser and curiouser: the role(s) of AID expression in self-tolerance.

Aicda is crucial for antibody diversification by mediating Ig class-switch recombination, V(D)J hypermutation (SHM) and, in some species, gene conversion. Recently, evidence has accumulated to show that Aicda is expressed during B-cell development and that this expression in some unknown way, mediates tolerance in immature and transitional B cells. In this issue of the European Journal of Immunology, Umiker et al. [Eur. J. Immunol. 2014. 44: 3093-3108] show that enforced expression of Aicda during early B-cell development is associated with self-tolerance. Curiously, constitutive Aicda expression that begins early in B cells suppresses the generation of autoreactive IgM but promotes the expression of self-reactive IgG. In contrast, when Aicda is activated later in B-cell development, self-reactive IgM is abundant but IgG is not. These observations suggest pathways for self-tolerance that have been little explored.

Authors
Kelsoe, G
MLA Citation
Kelsoe, G. "Curiouser and curiouser: the role(s) of AID expression in self-tolerance." European journal of immunology 44.10 (October 2014): 2876-2879.
PMID
25308427
Source
epmc
Published In
European Journal of Immunology
Volume
44
Issue
10
Publish Date
2014
Start Page
2876
End Page
2879
DOI
10.1002/eji.201445102

Immunoglobulin gene insertions and deletions in the affinity maturation of HIV-1 broadly reactive neutralizing antibodies.

Induction of HIV-1 broad neutralizing antibodies (bnAbs) is a goal of HIV-1 vaccine development but has remained challenging partially due to unusual traits of bnAbs, including high somatic hypermutation (SHM) frequencies and in-frame insertions and deletions (indels). Here we examined the propensity and functional requirement for indels within HIV-1 bnAbs. High-throughput sequencing of the immunoglobulin (Ig) VHDJH genes in HIV-1 infected and uninfected individuals revealed that the indel frequency was elevated among HIV-1-infected subjects, with no unique properties attributable to bnAb-producing individuals. This increased indel occurrence depended only on the frequency of SHM point mutations. Indel-encoded regions were generally proximal to antigen binding sites. Additionally, reconstruction of a HIV-1 CD4-binding site bnAb clonal lineage revealed that a large compound VHDJH indel was required for bnAb activity. Thus, vaccine development should focus on designing regimens targeted at sustained activation of bnAb lineages to achieve the required SHM and indel events.

Authors
Kepler, TB; Liao, H-X; Alam, SM; Bhaskarabhatla, R; Zhang, R; Yandava, C; Stewart, S; Anasti, K; Kelsoe, G; Parks, R; Lloyd, KE; Stolarchuk, C; Pritchett, J; Solomon, E; Friberg, E; Morris, L; Karim, SSA; Cohen, MS; Walter, E; Moody, MA; Wu, X; Altae-Tran, HR; Georgiev, IS; Kwong, PD; Boyd, SD; Fire, AZ; Mascola, JR; Haynes, BF
MLA Citation
Kepler, TB, Liao, H-X, Alam, SM, Bhaskarabhatla, R, Zhang, R, Yandava, C, Stewart, S, Anasti, K, Kelsoe, G, Parks, R, Lloyd, KE, Stolarchuk, C, Pritchett, J, Solomon, E, Friberg, E, Morris, L, Karim, SSA, Cohen, MS, Walter, E, Moody, MA, Wu, X, Altae-Tran, HR, Georgiev, IS, Kwong, PD, Boyd, SD, Fire, AZ, Mascola, JR, and Haynes, BF. "Immunoglobulin gene insertions and deletions in the affinity maturation of HIV-1 broadly reactive neutralizing antibodies." Cell host & microbe 16.3 (September 2014): 304-313.
PMID
25211073
Source
epmc
Published In
Cell Host & Microbe
Volume
16
Issue
3
Publish Date
2014
Start Page
304
End Page
313
DOI
10.1016/j.chom.2014.08.006

Cooperation of B cell lineages in induction of HIV-1-broadly neutralizing antibodies

Development of strategies for induction of HIV-1 broadly neutralizing antibodies (bnAbs) by vaccines is a priority. Determining the steps of bnAb induction in HIV-1-infected individuals who make bnAbs is a key strategy for immunogen design. Here, we study the B cell response in a bnAb-producing individual and report cooperation between two B cell lineages to drive bnAb development. We isolated a virus-neutralizing antibody lineage that targeted an envelope region (loop D) and selected virus escape mutants that resulted in both enhanced bnAb lineage envelope binding and escape mutant neutralization - traits associated with increased B cell antigen drive. Thus, in this individual, two B cell lineages cooperated to induce the development of bnAbs. Design of vaccine immunogens that simultaneously drive both helper and broadly neutralizing B cell lineages may be important for vaccine-induced recapitulation of events that transpire during the maturation of neutralizing antibodies in HIV-1-infected individuals. © 2014 Elsevier Inc.

Authors
Gao, F; Bonsignori, M; Liao, HX; Kumar, A; Xia, SM; Lu, X; Cai, F; Hwang, KK; Song, H; Zhou, T; Lynch, RM; Alam, SM; Moody, MA; Ferrari, G; Berrong, M; Kelsoe, G; Shaw, GM; Hahn, BH; Montefiori, DC; Kamanga, G; Cohen, MS; Hraber, P; Kwong, PD; Korber, BT; Mascola, JR; Kepler, TB; Haynes, BF
MLA Citation
Gao, F, Bonsignori, M, Liao, HX, Kumar, A, Xia, SM, Lu, X, Cai, F, Hwang, KK, Song, H, Zhou, T, Lynch, RM, Alam, SM, Moody, MA, Ferrari, G, Berrong, M, Kelsoe, G, Shaw, GM, Hahn, BH, Montefiori, DC, Kamanga, G, Cohen, MS, Hraber, P, Kwong, PD, Korber, BT, Mascola, JR, Kepler, TB, and Haynes, BF. "Cooperation of B cell lineages in induction of HIV-1-broadly neutralizing antibodies." Cell 158.3 (July 31, 2014): 481-491.
Source
scopus
Published In
Cell
Volume
158
Issue
3
Publish Date
2014
Start Page
481
End Page
491
DOI
10.1016/j.cell.2014.06.022

Cooperation of B cell lineages in induction of HIV-1-broadly neutralizing antibodies.

Development of strategies for induction of HIV-1 broadly neutralizing antibodies (bnAbs) by vaccines is a priority. Determining the steps of bnAb induction in HIV-1-infected individuals who make bnAbs is a key strategy for immunogen design. Here, we study the B cell response in a bnAb-producing individual and report cooperation between two B cell lineages to drive bnAb development. We isolated a virus-neutralizing antibody lineage that targeted an envelope region (loop D) and selected virus escape mutants that resulted in both enhanced bnAb lineage envelope binding and escape mutant neutralization-traits associated with increased B cell antigen drive. Thus, in this individual, two B cell lineages cooperated to induce the development of bnAbs. Design of vaccine immunogens that simultaneously drive both helper and broadly neutralizing B cell lineages may be important for vaccine-induced recapitulation of events that transpire during the maturation of neutralizing antibodies in HIV-1-infected individuals.

Authors
Gao, F; Bonsignori, M; Liao, H-X; Kumar, A; Xia, S-M; Lu, X; Cai, F; Hwang, K-K; Song, H; Zhou, T; Lynch, RM; Alam, SM; Moody, MA; Ferrari, G; Berrong, M; Kelsoe, G; Shaw, GM; Hahn, BH; Montefiori, DC; Kamanga, G; Cohen, MS; Hraber, P; Kwong, PD; Korber, BT; Mascola, JR; Kepler, TB; Haynes, BF
MLA Citation
Gao, F, Bonsignori, M, Liao, H-X, Kumar, A, Xia, S-M, Lu, X, Cai, F, Hwang, K-K, Song, H, Zhou, T, Lynch, RM, Alam, SM, Moody, MA, Ferrari, G, Berrong, M, Kelsoe, G, Shaw, GM, Hahn, BH, Montefiori, DC, Kamanga, G, Cohen, MS, Hraber, P, Kwong, PD, Korber, BT, Mascola, JR, Kepler, TB, and Haynes, BF. "Cooperation of B cell lineages in induction of HIV-1-broadly neutralizing antibodies." Cell 158.3 (July 24, 2014): 481-491.
PMID
25065977
Source
epmc
Published In
Cell
Volume
158
Issue
3
Publish Date
2014
Start Page
481
End Page
491
DOI
10.1016/j.cell.2014.06.022

Progress in HIV-1 vaccine development.

The past 2 years have seen a number of basic and translational science advances in the quest for development of an effective HIV-1 vaccine. These advances include discovery of new envelope targets of potentially protective antibodies, demonstration that CD8(+) T cells can control HIV-1 infection, development of immunogens to overcome HIV-1 T-cell epitope diversity, identification of correlates of transmission risk in an HIV-1 efficacy trial, and mapping of the coevolution of HIV-1 founder envelope mutants in infected subjects with broad neutralizing antibodies, thereby defining broad neutralizing antibody developmental pathways. Despite these advances, a promising HIV-1 vaccine efficacy trial published in 2013 did not prevent infection, and the HIV-1 vaccine field is still years away from deployment of an effective vaccine. This review summarizes what some of the scientific advances have been, what roadblocks still remain, and what the most promising approaches are for progress in design of successful HIV-1 vaccine candidates.

Authors
Haynes, BF; Moody, MA; Alam, M; Bonsignori, M; Verkoczy, L; Ferrari, G; Gao, F; Tomaras, GD; Liao, H-X; Kelsoe, G
MLA Citation
Haynes, BF, Moody, MA, Alam, M, Bonsignori, M, Verkoczy, L, Ferrari, G, Gao, F, Tomaras, GD, Liao, H-X, and Kelsoe, G. "Progress in HIV-1 vaccine development." The Journal of allergy and clinical immunology 134.1 (July 2014): 3-11. (Review)
PMID
25117798
Source
epmc
Published In
Journal of Allergy and Clinical Immunology
Volume
134
Issue
1
Publish Date
2014
Start Page
3
End Page
11
DOI
10.1016/j.jaci.2014.04.025

Redemption of autoreactive B cells.

Authors
Haynes, BF; Verkoczy, L; Kelsoe, G
MLA Citation
Haynes, BF, Verkoczy, L, and Kelsoe, G. "Redemption of autoreactive B cells." Proceedings of the National Academy of Sciences of the United States of America 111.25 (June 11, 2014): 9022-9023.
PMID
24920593
Source
epmc
Published In
Proceedings of the National Academy of Sciences of USA
Volume
111
Issue
25
Publish Date
2014
Start Page
9022
End Page
9023
DOI
10.1073/pnas.1407877111

HIV-1 envelope gp41 broadly neutralizing antibodies: hurdles for vaccine development.

Authors
Verkoczy, L; Kelsoe, G; Haynes, BF
MLA Citation
Verkoczy, L, Kelsoe, G, and Haynes, BF. "HIV-1 envelope gp41 broadly neutralizing antibodies: hurdles for vaccine development." PLoS pathogens 10.5 (May 22, 2014): e1004073-.
Website
http://hdl.handle.net/10161/10898
PMID
24853821
Source
epmc
Published In
PLoS pathogens
Volume
10
Issue
5
Publish Date
2014
Start Page
e1004073
DOI
10.1371/journal.ppat.1004073

Enhanced antibody responses to an HIV-1 membrane-proximal external region antigen in mice reconstituted with cultured lymphocytes.

We have shown that the protective HIV-1 Ab, 2F5, avidly reacts with a conserved mammalian self-Ag, kynureninase, and that the development of B cells specific for the 2F5 epitope is constrained by immunological tolerance. These observations suggest that the capacity to mount Ab responses to the 2F5 epitope is mitigated by tolerance, but such capacity may be latent in the pretolerance and/or anergic B cell pools. In this study, we use B cell tetramer reagents to track the frequencies of B cells that recognize the HIV-1 2F5 epitope (SP62): in C57BL/6 mice, SP62-binding transitional B cells are readily identified in bone marrow but are lost during subsequent development. Unsurprisingly then, immunization with SP62 immunogen does not elicit significant humoral responses in normal C57BL/6 mice. Reconstitution of Rag1(null) mice with normal congenic B cells that have matured in vitro restores the capacity to mount significant serum Ab and germinal center responses to this HIV-1 epitope. These B cell cultures are permissive for the development of autoreactive B cells and support the development of SP62-specific B cell compartments normally lost in 2F5 Ab knockin mice. The recovery of humoral responses to the 2F5/SP62 epitope of HIV-1 by reconstitution with B cells containing forbidden, autoreactive clones provides direct evidence that normal C57BL/6 mice latently possess the capacity to generate humoral responses to a conserved, neutralizing HIV-1 epitope.

Authors
Holl, TM; Yang, G; Kuraoka, M; Verkoczy, L; Alam, SM; Moody, MA; Haynes, BF; Kelsoe, G
MLA Citation
Holl, TM, Yang, G, Kuraoka, M, Verkoczy, L, Alam, SM, Moody, MA, Haynes, BF, and Kelsoe, G. "Enhanced antibody responses to an HIV-1 membrane-proximal external region antigen in mice reconstituted with cultured lymphocytes." Journal of immunology (Baltimore, Md. : 1950) 192.7 (April 2014): 3269-3279.
PMID
24591365
Source
epmc
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
192
Issue
7
Publish Date
2014
Start Page
3269
End Page
3279
DOI
10.4049/jimmunol.1302829

An autoreactive antibody from an SLE/HIV-1 individual broadly neutralizes HIV-1.

Broadly HIV-1-neutralizing antibodies (BnAbs) display one or more unusual traits, including a long heavy chain complementarity-determining region 3 (HCDR3), polyreactivity, and high levels of somatic mutations. These shared characteristics suggest that BnAb development might be limited by immune tolerance controls. It has been postulated that HIV-1-infected individuals with autoimmune disease and defective immune tolerance mechanisms may produce BnAbs more readily than those without autoimmune diseases. In this study, we identified an HIV-1-infected individual with SLE who exhibited controlled viral load (<5,000 copies/ml) in the absence of controlling HLA phenotypes and developed plasma HIV-1 neutralization breadth. We collected memory B cells from this individual and isolated a BnAb, CH98, that targets the CD4 binding site (CD4bs) of HIV-1 envelope glycoprotein 120 (gp120). CH98 bound to human antigens including dsDNA, which is specifically associated with SLE. Anti-dsDNA reactivity was also present in the patient's plasma. CH98 had a mutation frequency of 25% and 15% nt somatic mutations in the heavy and light chain variable domains, respectively, a long HCDR3, and a deletion in the light chain CDR1. The occurrence of anti-dsDNA reactivity by a HIV-1 CD4bs BnAb in an individual with SLE raises the possibility that some BnAbs and SLE-associated autoantibodies arise from similar pools of B cells.

Authors
Bonsignori, M; Wiehe, K; Grimm, SK; Lynch, R; Yang, G; Kozink, DM; Perrin, F; Cooper, AJ; Hwang, K-K; Chen, X; Liu, M; McKee, K; Parks, RJ; Eudailey, J; Wang, M; Clowse, M; Criscione-Schreiber, LG; Moody, MA; Ackerman, ME; Boyd, SD; Gao, F; Kelsoe, G; Verkoczy, L; Tomaras, GD; Liao, H-X; Kepler, TB; Montefiori, DC; Mascola, JR; Haynes, BF
MLA Citation
Bonsignori, M, Wiehe, K, Grimm, SK, Lynch, R, Yang, G, Kozink, DM, Perrin, F, Cooper, AJ, Hwang, K-K, Chen, X, Liu, M, McKee, K, Parks, RJ, Eudailey, J, Wang, M, Clowse, M, Criscione-Schreiber, LG, Moody, MA, Ackerman, ME, Boyd, SD, Gao, F, Kelsoe, G, Verkoczy, L, Tomaras, GD, Liao, H-X, Kepler, TB, Montefiori, DC, Mascola, JR, and Haynes, BF. "An autoreactive antibody from an SLE/HIV-1 individual broadly neutralizes HIV-1." The Journal of clinical investigation 124.4 (April 2014): 1835-1843.
PMID
24614107
Source
epmc
Published In
Journal of Clinical Investigation
Volume
124
Issue
4
Publish Date
2014
Start Page
1835
End Page
1843
DOI
10.1172/jci73441

Metabolic reprogramming is required for antibody production that is suppressed in anergic but exaggerated in chronically BAFF-exposed B cells.

B cell activation leads to proliferation and Ab production that can protect from pathogens or promote autoimmunity. Regulation of cell metabolism is essential to support the demands of lymphocyte growth and effector function and may regulate tolerance. In this study, we tested the regulation and role of glucose uptake and metabolism in the proliferation and Ab production of control, anergic, and autoimmune-prone B cells. Control B cells had a balanced increase in lactate production and oxygen consumption following activation, with proportionally increased glucose transporter Glut1 expression and mitochondrial mass upon either LPS or BCR stimulation. This contrasted with metabolic reprogramming of T cells, which had lower glycolytic flux when resting but disproportionately increased this pathway upon activation. Importantly, tolerance greatly affected B cell metabolic reprogramming. Anergic B cells remained metabolically quiescent, with only a modest increase in glycolysis and oxygen consumption with LPS stimulation. B cells chronically stimulated with elevated BAFF, however, rapidly increased glycolysis and Ab production upon stimulation. Induction of glycolysis was critical for Ab production, as glycolytic inhibition with the pyruvate dehydrogenase kinase inhibitor dichloroacetate sharply suppressed B cell proliferation and Ab secretion in vitro and in vivo. Furthermore, B cell-specific deletion of Glut1 led to reduced B cell numbers and impaired Ab production in vivo. Together, these data show that activated B cells require Glut1-dependent metabolic reprogramming to support proliferation and Ab production that is distinct from T cells and that this glycolytic reprogramming is regulated in tolerance.

Authors
Caro-Maldonado, A; Wang, R; Nichols, AG; Kuraoka, M; Milasta, S; Sun, LD; Gavin, AL; Abel, ED; Kelsoe, G; Green, DR; Rathmell, JC
MLA Citation
Caro-Maldonado, A, Wang, R, Nichols, AG, Kuraoka, M, Milasta, S, Sun, LD, Gavin, AL, Abel, ED, Kelsoe, G, Green, DR, and Rathmell, JC. "Metabolic reprogramming is required for antibody production that is suppressed in anergic but exaggerated in chronically BAFF-exposed B cells." Journal of immunology (Baltimore, Md. : 1950) 192.8 (April 2014): 3626-3636.
PMID
24616478
Source
epmc
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
192
Issue
8
Publish Date
2014
Start Page
3626
End Page
3636
DOI
10.4049/jimmunol.1302062

A tribute to Michael S. Neuberger.

Authors
Gearhart, PJ; Kelsoe, G
MLA Citation
Gearhart, PJ, and Kelsoe, G. "A tribute to Michael S. Neuberger." The Journal of clinical investigation 124.1 (January 2, 2014): 3-5.
PMID
24382382
Source
epmc
Published In
Journal of Clinical Investigation
Volume
124
Issue
1
Publish Date
2014
Start Page
3
End Page
5
DOI
10.1172/jci74366

IGHV1-69 B cell chronic lymphocytic leukemia antibodies cross-react with HIV-1 and hepatitis C virus antigens as well as intestinal commensal bacteria.

B-cell chronic lymphocytic leukemia (B-CLL) patients expressing unmutated immunoglobulin heavy variable regions (IGHVs) use the IGHV1-69 B cell receptor (BCR) in 25% of cases. Since HIV-1 envelope gp41 antibodies also frequently use IGHV1-69 gene segments, we hypothesized that IGHV1-69 B-CLL precursors may contribute to the gp41 B cell response during HIV-1 infection. To test this hypothesis, we rescued 5 IGHV1-69 unmutated antibodies as heterohybridoma IgM paraproteins and as recombinant IgG1 antibodies from B-CLL patients, determined their antigenic specificities and analyzed BCR sequences. IGHV1-69 B-CLL antibodies were enriched for reactivity with HIV-1 envelope gp41, influenza, hepatitis C virus E2 protein and intestinal commensal bacteria. These IGHV1-69 B-CLL antibodies preferentially used IGHD3 and IGHJ6 gene segments and had long heavy chain complementary determining region 3s (HCDR3s) (≥21 aa). IGHV1-69 B-CLL BCRs exhibited a phenylalanine at position 54 (F54) of the HCDR2 as do rare HIV-1 gp41 and influenza hemagglutinin stem neutralizing antibodies, while IGHV1-69 gp41 antibodies induced by HIV-1 infection predominantly used leucine (L54) allelic variants. These results demonstrate that the B-CLL cell population is an expansion of members of the innate polyreactive B cell repertoire with reactivity to a number of infectious agent antigens including intestinal commensal bacteria. The B-CLL IGHV1-69 B cell usage of F54 allelic variants strongly suggests that IGHV1-69 B-CLL gp41 antibodies derive from a restricted B cell pool that also produces rare HIV-1 gp41 and influenza hemagglutinin stem antibodies.

Authors
Hwang, K-K; Trama, AM; Kozink, DM; Chen, X; Wiehe, K; Cooper, AJ; Xia, S-M; Wang, M; Marshall, DJ; Whitesides, J; Alam, M; Tomaras, GD; Allen, SL; Rai, KR; McKeating, J; Catera, R; Yan, X-J; Chu, CC; Kelsoe, G; Liao, H-X; Chiorazzi, N; Haynes, BF
MLA Citation
Hwang, K-K, Trama, AM, Kozink, DM, Chen, X, Wiehe, K, Cooper, AJ, Xia, S-M, Wang, M, Marshall, DJ, Whitesides, J, Alam, M, Tomaras, GD, Allen, SL, Rai, KR, McKeating, J, Catera, R, Yan, X-J, Chu, CC, Kelsoe, G, Liao, H-X, Chiorazzi, N, and Haynes, BF. "IGHV1-69 B cell chronic lymphocytic leukemia antibodies cross-react with HIV-1 and hepatitis C virus antigens as well as intestinal commensal bacteria." PloS one 9.3 (January 2014): e90725-.
Website
http://hdl.handle.net/10161/10901
PMID
24614505
Source
epmc
Published In
PloS one
Volume
9
Issue
3
Publish Date
2014
Start Page
e90725
DOI
10.1371/journal.pone.0090725

Reconstructing a B-Cell Clonal Lineage. II. Mutation, Selection, and Affinity Maturation.

Affinity maturation of the antibody response is a fundamental process in adaptive immunity during which B-cells activated by infection or vaccination undergo rapid proliferation accompanied by the acquisition of point mutations in their rearranged immunoglobulin (Ig) genes and selection for increased affinity for the eliciting antigen. The rate of somatic hypermutation at any position within an Ig gene is known to depend strongly on the local DNA sequence, and Ig genes have region-specific codon biases that influence the local mutation rate within the gene resulting in increased differential mutability in the regions that encode the antigen-binding domains. We have isolated a set of clonally related natural Ig heavy chain-light chain pairs from an experimentally infected influenza patient, inferred the unmutated ancestral rearrangements and the maturation intermediates, and synthesized all the antibodies using recombinant methods. The lineage exhibits a remarkably uniform rate of improvement of the effective affinity to influenza hemagglutinin (HA) over evolutionary time, increasing 1000-fold overall from the unmutated ancestor to the best of the observed antibodies. Furthermore, analysis of selection reveals that selection and mutation bias were concordant even at the level of maturation to a single antigen. Substantial improvement in affinity to HA occurred along mutationally preferred paths in sequence space and was thus strongly facilitated by the underlying local codon biases.

Authors
Kepler, TB; Munshaw, S; Wiehe, K; Zhang, R; Yu, J-S; Woods, CW; Denny, TN; Tomaras, GD; Alam, SM; Moody, MA; Kelsoe, G; Liao, H-X; Haynes, BF
MLA Citation
Kepler, TB, Munshaw, S, Wiehe, K, Zhang, R, Yu, J-S, Woods, CW, Denny, TN, Tomaras, GD, Alam, SM, Moody, MA, Kelsoe, G, Liao, H-X, and Haynes, BF. "Reconstructing a B-Cell Clonal Lineage. II. Mutation, Selection, and Affinity Maturation." Frontiers in immunology 5 (January 2014): 170-.
Website
http://hdl.handle.net/10161/10899
PMID
24795717
Source
epmc
Published In
Frontiers in Immunology
Volume
5
Publish Date
2014
Start Page
170
DOI
10.3389/fimmu.2014.00170

Identification of a tissue-specific, C/EBPβ-dependent pathway of differentiation for murine peritoneal macrophages.

Macrophages and dendritic cells (DC) are distributed throughout the body and play important roles in pathogen detection and tissue homeostasis. In tissues, resident macrophages exhibit distinct phenotypes and activities, yet the transcriptional pathways that specify tissue-specific macrophages are largely unknown. We investigated the functions and origins of two peritoneal macrophage populations in mice: small and large peritoneal macrophages (SPM and LPM, respectively). SPM and LPM differ in their ability to phagocytose apoptotic cells, as well as in the production of cytokines in response to LPS. In steady-state conditions, SPM are sustained by circulating precursors, whereas LPM are maintained independently of hematopoiesis; however, both populations are replenished by bone marrow precursors following radiation injury. Transcription factor analysis revealed that SPM and LPM express abundant CCAAT/enhancer binding protein (C/EBP)-β. Cebpb(-/-) mice exhibit elevated numbers of SPM-like cells but lack functional LPM. Alveolar macrophages are also missing in Cebpb(-/-) mice, although macrophage populations in the spleen, kidney, skin, mesenteric lymph nodes, and liver are normal. Adoptive transfer of SPM into Cebpb(-/-) mice results in SPM differentiation into LPM, yet donor SPM do not generate LPM after transfer into C/EBPβ-sufficient mice, suggesting that endogenous LPM inhibit differentiation by SPM. We conclude that C/EBPβ plays an intrinsic, tissue-restricted role in the generation of resident macrophages.

Authors
Cain, DW; O'Koren, EG; Kan, MJ; Womble, M; Sempowski, GD; Hopper, K; Gunn, MD; Kelsoe, G
MLA Citation
Cain, DW, O'Koren, EG, Kan, MJ, Womble, M, Sempowski, GD, Hopper, K, Gunn, MD, and Kelsoe, G. "Identification of a tissue-specific, C/EBPβ-dependent pathway of differentiation for murine peritoneal macrophages." J Immunol 191.9 (November 1, 2013): 4665-4675.
PMID
24078688
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
191
Issue
9
Publish Date
2013
Start Page
4665
End Page
4675
DOI
10.4049/jimmunol.1300581

The human fetal lymphocyte lineage: identification by CD27 and LIN28B expression in B cell progenitors.

CD27, a member of the TNFR superfamily, is used to identify human memory B cells. Nonetheless, CD27(+) B cells are present in patients with HIGM1 syndrome who are unable to generate GCs or memory B cells. CD27(+)IgD(+) fetal B cells are present in umbilical cord blood, and CD27 may also be a marker of the human B1-like B cells. To define the origin of naïve CD27(+)IgD(+) human B cells, we studied B cell development in both fetal and adult tissues. In human FL, most CD19(+) cells coexpressed CD10, a marker of human developing B cells. Some CD19(+)CD10(+) B cells expressed CD27, and these fetal CD27(+) cells were present in the pro-B, pre-B, and immature/transitional B cell compartments. Lower frequencies of phenotypically identical cells were also identified in adult BM. CD27(+) pro-B, pre-B, and immature/transitional B cells expressed recombination activating gene-1, terminal deoxynucleotidyl transferase and Vpre-B mRNA comparably to their CD27(-) counterparts. CD27(+) and CD27(-) developing B cells showed similar Ig heavy chain gene usage with low levels of mutations, suggesting that CD27(+) developing B cells are distinct from mutated memory B cells. Despite these similarities, CD27(+) developing B cells differed from CD27(-) developing B cells by their increased expression of LIN28B, a transcription factor associated with the fetal lymphoid lineages of mice. Furthermore, CD27(+) pro-B cells efficiently generated IgM(+)IgD(+) immature/transitional B cells in vitro. Our observations suggest that CD27 expression during B cell development identifies a physiologic state or lineage for human B cell development distinct from the memory B cell compartment.

Authors
McWilliams, L; Su, K-Y; Liang, X; Liao, D; Floyd, S; Amos, J; Moody, MA; Kelsoe, G; Kuraoka, M
MLA Citation
McWilliams, L, Su, K-Y, Liang, X, Liao, D, Floyd, S, Amos, J, Moody, MA, Kelsoe, G, and Kuraoka, M. "The human fetal lymphocyte lineage: identification by CD27 and LIN28B expression in B cell progenitors." J Leukoc Biol 94.5 (November 2013): 991-1001.
PMID
23901121
Source
pubmed
Published In
Journal of leukocyte biology
Volume
94
Issue
5
Publish Date
2013
Start Page
991
End Page
1001
DOI
10.1189/jlb.0113048

Co-evolution of a broadly neutralizing HIV-1 antibody and founder virus.

Current human immunodeficiency virus-1 (HIV-1) vaccines elicit strain-specific neutralizing antibodies. However, cross-reactive neutralizing antibodies arise in approximately 20% of HIV-1-infected individuals, and details of their generation could provide a blueprint for effective vaccination. Here we report the isolation, evolution and structure of a broadly neutralizing antibody from an African donor followed from the time of infection. The mature antibody, CH103, neutralized approximately 55% of HIV-1 isolates, and its co-crystal structure with the HIV-1 envelope protein gp120 revealed a new loop-based mechanism of CD4-binding-site recognition. Virus and antibody gene sequencing revealed concomitant virus evolution and antibody maturation. Notably, the unmutated common ancestor of the CH103 lineage avidly bound the transmitted/founder HIV-1 envelope glycoprotein, and evolution of antibody neutralization breadth was preceded by extensive viral diversification in and near the CH103 epitope. These data determine the viral and antibody evolution leading to induction of a lineage of HIV-1 broadly neutralizing antibodies, and provide insights into strategies to elicit similar antibodies by vaccination.

Authors
Liao, H-X; Lynch, R; Zhou, T; Gao, F; Alam, SM; Boyd, SD; Fire, AZ; Roskin, KM; Schramm, CA; Zhang, Z; Zhu, J; Shapiro, L; NISC Comparative Sequencing Program, ; Mullikin, JC; Gnanakaran, S; Hraber, P; Wiehe, K; Kelsoe, G; Yang, G; Xia, S-M; Montefiori, DC; Parks, R; Lloyd, KE; Scearce, RM; Soderberg, KA; Cohen, M; Kamanga, G; Louder, MK; Tran, LM; Chen, Y; Cai, F; Chen, S; Moquin, S; Du, X; Joyce, MG; Srivatsan, S; Zhang, B; Zheng, A; Shaw, GM; Hahn, BH; Kepler, TB; Korber, BTM; Kwong, PD et al.
MLA Citation
Liao, H-X, Lynch, R, Zhou, T, Gao, F, Alam, SM, Boyd, SD, Fire, AZ, Roskin, KM, Schramm, CA, Zhang, Z, Zhu, J, Shapiro, L, NISC Comparative Sequencing Program, , Mullikin, JC, Gnanakaran, S, Hraber, P, Wiehe, K, Kelsoe, G, Yang, G, Xia, S-M, Montefiori, DC, Parks, R, Lloyd, KE, Scearce, RM, Soderberg, KA, Cohen, M, Kamanga, G, Louder, MK, Tran, LM, Chen, Y, Cai, F, Chen, S, Moquin, S, Du, X, Joyce, MG, Srivatsan, S, Zhang, B, Zheng, A, Shaw, GM, Hahn, BH, Kepler, TB, Korber, BTM, and Kwong, PD et al. "Co-evolution of a broadly neutralizing HIV-1 antibody and founder virus." Nature 496.7446 (April 25, 2013): 469-476.
Website
http://hdl.handle.net/10161/10902
PMID
23552890
Source
pubmed
Published In
Nature
Volume
496
Issue
7446
Publish Date
2013
Start Page
469
End Page
476
DOI
10.1038/nature12053

Identification of autoantigens recognized by the 2F5 and 4E10 broadly neutralizing HIV-1 antibodies.

Many human monoclonal antibodies that neutralize multiple clades of HIV-1 are polyreactive and bind avidly to mammalian autoantigens. Indeed, the generation of neutralizing antibodies to the 2F5 and 4E10 epitopes of HIV-1 gp41 in man may be proscribed by immune tolerance because mice expressing the V(H) and V(L) regions of 2F5 have a block in B cell development that is characteristic of central tolerance. This developmental blockade implies the presence of tolerizing autoantigens that are mimicked by the membrane-proximal external region of HIV-1 gp41. We identify human kynureninase (KYNU) and splicing factor 3b subunit 3 (SF3B3) as the primary conserved, vertebrate self-antigens recognized by the 2F5 and 4E10 antibodies, respectively. 2F5 binds the H4 domain of KYNU which contains the complete 2F5 linear epitope (ELDKWA). 4E10 recognizes an epitope of SF3B3 that is strongly dependent on hydrophobic interactions. Opossums carry a rare KYNU H4 domain that abolishes 2F5 binding, but they retain the SF3B3 4E10 epitope. Immunization of opossums with HIV-1 gp140 induced extraordinary titers of serum antibody to the 2F5 ELDKWA epitope but little or nothing to the 4E10 determinant. Identification of structural motifs shared by vertebrates and HIV-1 provides direct evidence that immunological tolerance can impair humoral responses to HIV-1.

Authors
Yang, G; Holl, TM; Liu, Y; Li, Y; Lu, X; Nicely, NI; Kepler, TB; Alam, SM; Liao, H-X; Cain, DW; Spicer, L; VandeBerg, JL; Haynes, BF; Kelsoe, G
MLA Citation
Yang, G, Holl, TM, Liu, Y, Li, Y, Lu, X, Nicely, NI, Kepler, TB, Alam, SM, Liao, H-X, Cain, DW, Spicer, L, VandeBerg, JL, Haynes, BF, and Kelsoe, G. "Identification of autoantigens recognized by the 2F5 and 4E10 broadly neutralizing HIV-1 antibodies." J Exp Med 210.2 (February 11, 2013): 241-256.
Website
http://hdl.handle.net/10161/10900
PMID
23359068
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
210
Issue
2
Publish Date
2013
Start Page
241
End Page
256
DOI
10.1084/jem.20121977

Immunogenicity of membrane-bound HIV-1 gp41 membrane-proximal external region (MPER) segments is dominated by residue accessibility and modulated by stereochemistry

Structural characterization of epitope-paratope pairs has contributed to the understanding of antigenicity. By contrast, few structural studies relate to immunogenicity, the process of antigen-induced immune responses in vivo. Using a lipid-arrayed membrane-proximal external region (MPER) of HIV-1 glycoprotein 41 as a model antigen, we investigated the influence of physicochemical properties on immunogenicity in relation to structural modifications of MPER/liposome vaccines. Anchoring the MPER to the membrane via an alkyl tail or transmembrane domain retained the MPER on liposomes in vivo, while preserving MPER secondary structure. However, structural modifications that affected MPER membrane orientation and antigenic residue accessibility strongly impacted induced antibody responses. The solvent-exposed MPER tryptophan residue (Trp-680) was immunodominant, focusing immune responses, despite sequence variability elsewhere. Nonetheless, immunogenicity could be readily manipulated using site-directed mutagenesis or structural constraints tomodulate amino acid surface display. These studies provide fundamental insights for immunogen design aimed at targeting B cell antibody responses. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc.

Authors
Kim, M; Song, L; Moon, J; Sun, Z-YJ; Bershteyn, A; Hanson, M; Cain, D; Goka, S; Kelsoe, G; Wagner, G; al, E
MLA Citation
Kim, M, Song, L, Moon, J, Sun, Z-YJ, Bershteyn, A, Hanson, M, Cain, D, Goka, S, Kelsoe, G, Wagner, G, and al, E. "Immunogenicity of membrane-bound HIV-1 gp41 membrane-proximal external region (MPER) segments is dominated by residue accessibility and modulated by stereochemistry." Journal of Biological Chemistry 288.44 (2013): 31888-31901.
PMID
24047898
Source
scival
Published In
The Journal of biological chemistry
Volume
288
Issue
44
Publish Date
2013
Start Page
31888
End Page
31901
DOI
10.1074/jbc.M113.494609

Disparate adjuvant properties among three formulations of " alum"

Aluminum adjuvants, commonly referred to as " alum," are the most widespread immunostimulants in human vaccines. Although the mechanisms that promote humoral responses to alum-adsorbed antigens are still enigmatic, alum is thought to form antigen depots and induce inflammatory signals that, in turn, promote antibody production. It was recently noted that Imject® alum, a commercial aluminum-containing adjuvant commonly used in animal studies, is not the physicochemical equivalent of aluminum adjuvant present in human vaccines. This difference raises concerns about the use of Imject® alum in animal research as a model for approved aluminum adjuvants. Here, we compared the capacity of Imject® alum, Alhydrogel®, and a traditional alum-antigen precipitate to induce humoral responses in mice to the hapten-carrier antigen, NP-CGG [(4-hydroxy-3-nitrophenyl)acetyl-chicken γ-globulin]. The magnitude of humoral responses elicited by Alhydrogel® and precipitated alum was significantly greater than that induced by Imject® alum. The strength of the humoral responses elicited by different alum formulations was correlated with the quantity of pro-inflammatory cytokines induced and the numbers of inflammatory cells at the site of immunization. Moreover, Imject® exhibited a severely reduced capacity to adsorb protein antigens compared to Alhydrogel® and precipitated alum. These findings reveal substantial differences in the immunostimulatory properties of distinct alum preparations, an important point of consideration for the evaluation of novel adjuvants, the assessment of new alum-based vaccines, and in mechanistic studies of adjuvanticity. © 2012 Elsevier Ltd.

Authors
Cain, DW; Sanders, SE; Cunningham, MM; Kelsoe, G
MLA Citation
Cain, DW, Sanders, SE, Cunningham, MM, and Kelsoe, G. "Disparate adjuvant properties among three formulations of " alum"." Vaccine 31.4 (2013): 653-660.
Source
scival
Published In
Vaccine
Volume
31
Issue
4
Publish Date
2013
Start Page
653
End Page
660
DOI
10.1016/j.vaccine.2012.11.044

HIV-1 gp120 vaccine induces affinity maturation in both new and persistent antibody clonal lineages.

Most antibodies that broadly neutralize HIV-1 are highly somatically mutated in antibody clonal lineages that persist over time. Here, we describe the analysis of human antibodies induced during an HIV-1 vaccine trial (GSK PRO HIV-002) that used the clade B envelope (Env) gp120 of clone W6.1D (gp120(W6.1D)). Using dual-color antigen-specific sorting, we isolated Env-specific human monoclonal antibodies (MAbs) and studied the clonal persistence of antibodies in the setting of HIV-1 Env vaccination. We found evidence of V(H) somatic mutation induced by the vaccine but only to a modest level (3.8% ± 0.5%; range 0 to 8.2%). Analysis of 34 HIV-1-reactive MAbs recovered over four immunizations revealed evidence of both sequential recruitment of naïve B cells and restimulation of previously recruited memory B cells. These recombinant antibodies recapitulated the anti-HIV-1 activity of participant serum including pseudovirus neutralization and antibody-dependent cell-mediated cytotoxicity (ADCC). One antibody (3491) demonstrated a change in specificity following somatic mutation with binding of the inferred unmutated ancestor to a linear C2 peptide while the mutated antibody reacted only with a conformational epitope in gp120 Env. Thus, gp120(W6.1D) was strongly immunogenic but over four immunizations induced levels of affinity maturation below that of broadly neutralizing MAbs. Improved vaccination strategies will be needed to drive persistent stimulation of antibody clonal lineages to induce affinity maturation that results in highly mutated HIV-1 Env-reactive antibodies.

Authors
Moody, MA; Yates, NL; Amos, JD; Drinker, MS; Eudailey, JA; Gurley, TC; Marshall, DJ; Whitesides, JF; Chen, X; Foulger, A; Yu, J-S; Zhang, R; Meyerhoff, RR; Parks, R; Scull, JC; Wang, L; Vandergrift, NA; Pickeral, J; Pollara, J; Kelsoe, G; Alam, SM; Ferrari, G; Montefiori, DC; Voss, G; Liao, H-X; Tomaras, GD; Haynes, BF
MLA Citation
Moody, MA, Yates, NL, Amos, JD, Drinker, MS, Eudailey, JA, Gurley, TC, Marshall, DJ, Whitesides, JF, Chen, X, Foulger, A, Yu, J-S, Zhang, R, Meyerhoff, RR, Parks, R, Scull, JC, Wang, L, Vandergrift, NA, Pickeral, J, Pollara, J, Kelsoe, G, Alam, SM, Ferrari, G, Montefiori, DC, Voss, G, Liao, H-X, Tomaras, GD, and Haynes, BF. "HIV-1 gp120 vaccine induces affinity maturation in both new and persistent antibody clonal lineages." J Virol 86.14 (July 2012): 7496-7507.
PMID
22553329
Source
pubmed
Published In
Journal of virology
Volume
86
Issue
14
Publish Date
2012
Start Page
7496
End Page
7507
DOI
10.1128/JVI.00426-12

B-cell-lineage immunogen design in vaccine development with HIV-1 as a case study.

Failure of immunization with the HIV-1 envelope to induce broadly neutralizing antibodies against conserved epitopes is a major barrier to producing a preventive HIV-1 vaccine. Broadly neutralizing monoclonal antibodies (BnAbs) from those subjects who do produce them after years of chronic HIV-1 infection have one or more unusual characteristics, including polyreactivity for host antigens, extensive somatic hypermutation and long, variable heavy-chain third complementarity-determining regions, factors that may limit their expression by host immunoregulatory mechanisms. The isolation of BnAbs from HIV-1-infected subjects and the use of computationally derived clonal lineages as templates provide a new path for HIV-1 vaccine immunogen design. This approach, which should be applicable to many infectious agents, holds promise for the construction of vaccines that can drive B cells along rare but desirable maturation pathways.

Authors
Haynes, BF; Kelsoe, G; Harrison, SC; Kepler, TB
MLA Citation
Haynes, BF, Kelsoe, G, Harrison, SC, and Kepler, TB. "B-cell-lineage immunogen design in vaccine development with HIV-1 as a case study. (Published online)" Nat Biotechnol 30.5 (May 7, 2012): 423-433.
PMID
22565972
Source
pubmed
Published In
Nature Biotechnology
Volume
30
Issue
5
Publish Date
2012
Start Page
423
End Page
433
DOI
10.1038/nbt.2197

Isolation of HIV-1-neutralizing mucosal monoclonal antibodies from human colostrum.

BACKGROUND: Generation of potent anti-HIV antibody responses in mucosal compartments is a potential requirement of a transmission-blocking HIV vaccine. HIV-specific, functional antibody responses are present in breast milk, and these mucosal antibody responses may play a role in protection of the majority of HIV-exposed, breastfeeding infants. Therefore, characterization of HIV-specific antibodies produced by B cells in milk could guide the development of vaccines that elicit protective mucosal antibody responses. METHODS: We isolated B cells from colostrum of an HIV-infected lactating woman with a detectable neutralization response in milk and recombinantly produced and characterized the resulting HIV-1 Envelope (Env)-specific monoclonal antibodies (mAbs). RESULTS: The identified HIV-1 Env-specific colostrum mAbs, CH07 and CH08, represent two of the first mucosally-derived anti-HIV antibodies yet to be reported. Colostrum mAb CH07 is a highly-autoreactive, weakly-neutralizing gp140-specific mAb that binds to linear epitopes in the gp120 C5 region and gp41 fusion domain. In contrast, colostrum mAb CH08 is a nonpolyreactive CD4-inducible (CD4i) gp120-specific mAb with moderate breadth of neutralization. CONCLUSIONS: These novel HIV-neutralizing mAbs isolated from a mucosal compartment provide insight into the ability of mucosal B cell populations to produce functional anti-HIV antibodies that may contribute to protection against virus acquisition at mucosal surfaces.

Authors
Friedman, J; Alam, SM; Shen, X; Xia, S-M; Stewart, S; Anasti, K; Pollara, J; Fouda, GG; Yang, G; Kelsoe, G; Ferrari, G; Tomaras, GD; Haynes, BF; Liao, H-X; Moody, MA; Permar, SR
MLA Citation
Friedman, J, Alam, SM, Shen, X, Xia, S-M, Stewart, S, Anasti, K, Pollara, J, Fouda, GG, Yang, G, Kelsoe, G, Ferrari, G, Tomaras, GD, Haynes, BF, Liao, H-X, Moody, MA, and Permar, SR. "Isolation of HIV-1-neutralizing mucosal monoclonal antibodies from human colostrum." PLoS One 7.5 (2012): e37648-.
Website
http://hdl.handle.net/10161/10587
PMID
22624058
Source
pubmed
Published In
PloS one
Volume
7
Issue
5
Publish Date
2012
Start Page
e37648
DOI
10.1371/journal.pone.0037648

Disparate adjuvant properties among three formulations of "alum"

Aluminum adjuvants, commonly referred to as "alum," are the most widespread immunostimulants in human vaccines. Although the mechanisms that promote humoral responses to alum-adsorbed antigens are still enigmatic, alum is thought to form antigen depots and induce inflammatory signals that, in turn, promote antibody production. It was recently noted that Imject® alum, a commercial aluminum-containing adjuvant commonly used in animal studies, is not the physicochemical equivalent of aluminum adjuvant present in human vaccines. This difference raises concerns about the use of Imject® alum in animal research as a model for approved aluminum adjuvants. Here, we compared the capacity of Imject® alum, Alhydrogel®, and a traditional alum-antigen precipitate to induce humoral responses in mice to the hapten-carrier antigen, NP-CGG [(4-hydroxy-3-nitrophenyl)acetyl-chicken γ-globulin]. The magnitude of humoral responses elicited by Alhydrogel® and precipitated alum was significantly greater than that induced by Imject® alum. The strength of the humoral responses elicited by different alum formulations was correlated with the quantity of pro-inflammatory cytokines induced and the numbers of inflammatory cells at the site of immunization. Moreover, Imject® exhibited a severely reduced capacity to adsorb protein antigens compared to Alhydrogel® and precipitated alum. These findings reveal substantial differences in the immunostimulatory properties of distinct alum preparations, an important point of consideration for the evaluation of novel adjuvants, the assessment of new alum-based vaccines, and in mechanistic studies of adjuvanticity. © 2012 Elsevier Ltd. All rights reserved.

Authors
Cain, DW; Sanders, SE; Cunningham, MM; Kelsoe, G
MLA Citation
Cain, DW, Sanders, SE, Cunningham, MM, and Kelsoe, G. "Disparate adjuvant properties among three formulations of "alum"." Vaccine (2012).
PMID
23200935
Source
scival
Published In
Vaccine
Publish Date
2012
DOI
10.1016/j.vaccine.2012.11.044

Differential reactivity of germ line allelic variants of a broadly neutralizing HIV-1 antibody to a gp41 fusion intermediate conformation.

Genetic factors, as well as antigenic stimuli, can influence antibody repertoire formation. Moreover, the affinity of antigen for unmutated naïve B cell receptors determines the threshold for activation of germinal center antibody responses. The gp41 2F5 broadly neutralizing antibody (bNAb) uses the V(H)2-5 gene, which has 10 distinct alleles that use either a heavy-chain complementarity-determining region 2 (HCDR2) aspartic acid (D(H54)) or an HCDR2 asparagine (N(H54)) residue. The 2F5 HCDR2 D(H54) residue has been shown to form a salt bridge with gp41 (665)K; the V(H)2-5 germ line allele variant containing N(H54) cannot do so and thus should bind less avidly to gp41. Thus, the induction of 2F5 bNAb is dependent on both genetic and structural factors that could affect antigen affinity of unmutated naïve B cell receptors. Here, we studied allelic variants of the V(H)2-5 inferred germ line forms of the HIV-1 gp41 bNAb 2F5 for their antigen binding affinities to gp41 linear peptide and conformational protein antigens. Both V(H)2-5 2F5 inferred germ line variants bound to gp41 peptides and protein, including the fusion intermediate protein mimic, although more weakly than the mature 2F5 antibody. As predicted, the affinity of the N(H54) variant for fusion-intermediate conformation was an order of magnitude lower than that of the D(H54) V(H)2-5 germ line antibody, demonstrating that allelic variants of 2F5 germ line antibodies differentially bind to gp41. Thus, these data demonstrate a genetically determined trait that may affect host responses to HIV-1 envelope epitopes recognized by broadly neutralizing antibodies and has implications for unmutated ancestor-based immunogen design.

Authors
Alam, SM; Liao, H-X; Dennison, SM; Jaeger, F; Parks, R; Anasti, K; Foulger, A; Donathan, M; Lucas, J; Verkoczy, L; Nicely, N; Tomaras, GD; Kelsoe, G; Chen, B; Kepler, TB; Haynes, BF
MLA Citation
Alam, SM, Liao, H-X, Dennison, SM, Jaeger, F, Parks, R, Anasti, K, Foulger, A, Donathan, M, Lucas, J, Verkoczy, L, Nicely, N, Tomaras, GD, Kelsoe, G, Chen, B, Kepler, TB, and Haynes, BF. "Differential reactivity of germ line allelic variants of a broadly neutralizing HIV-1 antibody to a gp41 fusion intermediate conformation." J Virol 85.22 (November 2011): 11725-11731.
PMID
21917975
Source
pubmed
Published In
Journal of virology
Volume
85
Issue
22
Publish Date
2011
Start Page
11725
End Page
11731
DOI
10.1128/JVI.05680-11

Initial antibodies binding to HIV-1 gp41 in acutely infected subjects are polyreactive and highly mutated.

The initial antibody response to HIV-1 is targeted to envelope (Env) gp41, and is nonneutralizing and ineffective in controlling viremia. To understand the origins and characteristics of gp41-binding antibodies produced shortly after HIV-1 transmission, we isolated and studied gp41-reactive plasma cells from subjects acutely infected with HIV-1. The frequencies of somatic mutations were relatively high in these gp41-reactive antibodies. Reverted unmutated ancestors of gp41-reactive antibodies derived from subjects acutely infected with HIV-1 frequently did not react with autologous HIV-1 Env; however, these antibodies were polyreactive and frequently bound to host or bacterial antigens. In one large clonal lineage of gp41-reactive antibodies, reactivity to HIV-1 Env was acquired only after somatic mutations. Polyreactive gp41-binding antibodies were also isolated from uninfected individuals. These data suggest that the majority of gp41-binding antibodies produced after acute HIV-1 infection are cross-reactive responses generated by stimulating memory B cells that have previously been activated by non-HIV-1 antigens.

Authors
Liao, H-X; Chen, X; Munshaw, S; Zhang, R; Marshall, DJ; Vandergrift, N; Whitesides, JF; Lu, X; Yu, J-S; Hwang, K-K; Gao, F; Markowitz, M; Heath, SL; Bar, KJ; Goepfert, PA; Montefiori, DC; Shaw, GC; Alam, SM; Margolis, DM; Denny, TN; Boyd, SD; Marshal, E; Egholm, M; Simen, BB; Hanczaruk, B; Fire, AZ; Voss, G; Kelsoe, G; Tomaras, GD; Moody, MA; Kepler, TB; Haynes, BF
MLA Citation
Liao, H-X, Chen, X, Munshaw, S, Zhang, R, Marshall, DJ, Vandergrift, N, Whitesides, JF, Lu, X, Yu, J-S, Hwang, K-K, Gao, F, Markowitz, M, Heath, SL, Bar, KJ, Goepfert, PA, Montefiori, DC, Shaw, GC, Alam, SM, Margolis, DM, Denny, TN, Boyd, SD, Marshal, E, Egholm, M, Simen, BB, Hanczaruk, B, Fire, AZ, Voss, G, Kelsoe, G, Tomaras, GD, Moody, MA, Kepler, TB, and Haynes, BF. "Initial antibodies binding to HIV-1 gp41 in acutely infected subjects are polyreactive and highly mutated." J Exp Med 208.11 (October 24, 2011): 2237-2249.
PMID
21987658
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
208
Issue
11
Publish Date
2011
Start Page
2237
End Page
2249
DOI
10.1084/jem.20110363

A novel role for activation-induced cytidine deaminase: central B-cell tolerance.

Authors
Kuraoka, M; Kelsoe, G
MLA Citation
Kuraoka, M, and Kelsoe, G. "A novel role for activation-induced cytidine deaminase: central B-cell tolerance." Cell Cycle 10.20 (October 15, 2011): 3423-3424.
PMID
22067654
Source
pubmed
Published In
Cell Cycle
Volume
10
Issue
20
Publish Date
2011
Start Page
3423
End Page
3424
DOI
10.4161/cc.10.20.17693

Rescue of HIV-1 broad neutralizing antibody-expressing B cells in 2F5 VH x VL knockin mice reveals multiple tolerance controls.

The HIV-1 broadly neutralizing Ab (bnAb) 2F5 has been shown to be poly-/self-reactive in vitro, and we previously demonstrated that targeted expression of its VDJ rearrangement alone was sufficient to trigger a profound B cell developmental blockade in 2F5 V(H) knockin (KI) mice, consistent with central deletion of 2F5 H chain-expressing B cells. In this study, we generate a strain expressing the entire 2F5 bnAb specificity, 2F5 V(H) × V(L) KI mice, and find an even higher degree of tolerance control than observed in the 2F5 V(H) KI strain. Although B cell development was severely impaired in 2F5 V(H) × V(L) KI animals, we demonstrate rescue of their B cells when cultured in IL-7/BAFF. Intriguingly, even under these conditions, most rescued B cell hybridomas produced mAbs that lacked HIV-1 Envelope (Env) reactivity due to editing of the 2F5 L chain, and the majority of rescued B cells retained an anergic phenotype. Thus, when clonal deletion is circumvented, κ editing and anergy are additional safeguards preventing 2F5 V(H)/V(L) expression by immature/transitional B cells. Importantly, 7% of rescued B cells retained 2F5 V(H)/V(L) expression and secreted Env-specific mAbs with HIV-1-neutralizing activity. This partial rescue was further corroborated in vivo, as reflected by the anergic phenotype of most rescued B cells in 2F5 V(H) × V(L) KI × Eμ-Bcl-2 transgenic mice and significant (yet modest) enrichment of Env-specific B cells and serum Igs. The rescued 2F5 mAb-producing B cell clones in this study are the first examples, to our knowledge, of in vivo-derived bone marrow precursors specifying HIV-1 bnAbs and provide a starting point for design of strategies aimed at rescuing such B cells.

Authors
Verkoczy, L; Chen, Y; Bouton-Verville, H; Zhang, J; Diaz, M; Hutchinson, J; Ouyang, Y-B; Alam, SM; Holl, TM; Hwang, K-K; Kelsoe, G; Haynes, BF
MLA Citation
Verkoczy, L, Chen, Y, Bouton-Verville, H, Zhang, J, Diaz, M, Hutchinson, J, Ouyang, Y-B, Alam, SM, Holl, TM, Hwang, K-K, Kelsoe, G, and Haynes, BF. "Rescue of HIV-1 broad neutralizing antibody-expressing B cells in 2F5 VH x VL knockin mice reveals multiple tolerance controls." J Immunol 187.7 (October 1, 2011): 3785-3797.
PMID
21908739
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
187
Issue
7
Publish Date
2011
Start Page
3785
End Page
3797
DOI
10.4049/jimmunol.1101633

Analysis of a clonal lineage of HIV-1 envelope V2/V3 conformational epitope-specific broadly neutralizing antibodies and their inferred unmutated common ancestors.

V2/V3 conformational epitope antibodies that broadly neutralize HIV-1 (PG9 and PG16) have been recently described. Since an elicitation of previously known broadly neutralizing antibodies has proven elusive, the induction of antibodies with such specificity is an important goal for HIV-1 vaccine development. A critical question is which immunogens and vaccine formulations might be used to trigger and drive the development of memory B cell precursors with V2/V3 conformational epitope specificity. In this paper we identified a clonal lineage of four V2/V3 conformational epitope broadly neutralizing antibodies (CH01 to CH04) from an African HIV-1-infected broad neutralizer and inferred their common reverted unmutated ancestor (RUA) antibodies. While conformational epitope antibodies rarely bind recombinant Env monomers, a screen of 32 recombinant envelopes for binding to the CH01 to CH04 antibodies showed monoclonal antibody (MAb) binding to the E.A244 gp120 Env and to chronic Env AE.CM243; MAbs CH01 and CH02 also bound to transmitted/founder Env B.9021. CH01 to CH04 neutralized 38% to 49% of a panel of 91 HIV-1 tier 2 pseudoviruses, while the RUAs neutralized only 16% of HIV-1 isolates. Although the reverted unmutated ancestors showed restricted neutralizing activity, they retained the ability to bind to the E.A244 gp120 HIV-1 envelope with an affinity predicted to trigger B cell development. Thus, E.A244, B.9021, and AE.CM243 Envs are three potential immunogen candidates for studies aimed at defining strategies to induce V2/V3 conformational epitope-specific antibodies.

Authors
Bonsignori, M; Hwang, K-K; Chen, X; Tsao, C-Y; Morris, L; Gray, E; Marshall, DJ; Crump, JA; Kapiga, SH; Sam, NE; Sinangil, F; Pancera, M; Yongping, Y; Zhang, B; Zhu, J; Kwong, PD; O'Dell, S; Mascola, JR; Wu, L; Nabel, GJ; Phogat, S; Seaman, MS; Whitesides, JF; Moody, MA; Kelsoe, G; Yang, X; Sodroski, J; Shaw, GM; Montefiori, DC; Kepler, TB; Tomaras, GD; Alam, SM; Liao, H-X; Haynes, BF
MLA Citation
Bonsignori, M, Hwang, K-K, Chen, X, Tsao, C-Y, Morris, L, Gray, E, Marshall, DJ, Crump, JA, Kapiga, SH, Sam, NE, Sinangil, F, Pancera, M, Yongping, Y, Zhang, B, Zhu, J, Kwong, PD, O'Dell, S, Mascola, JR, Wu, L, Nabel, GJ, Phogat, S, Seaman, MS, Whitesides, JF, Moody, MA, Kelsoe, G, Yang, X, Sodroski, J, Shaw, GM, Montefiori, DC, Kepler, TB, Tomaras, GD, Alam, SM, Liao, H-X, and Haynes, BF. "Analysis of a clonal lineage of HIV-1 envelope V2/V3 conformational epitope-specific broadly neutralizing antibodies and their inferred unmutated common ancestors." J Virol 85.19 (October 2011): 9998-10009.
PMID
21795340
Source
pubmed
Published In
Journal of virology
Volume
85
Issue
19
Publish Date
2011
Start Page
9998
End Page
10009
DOI
10.1128/JVI.05045-11

Activation-induced cytidine deaminase mediates central tolerance in B cells.

The Aicda gene product, activation-induced cytidine deaminase (AID), initiates somatic hypermutation, class-switch recombination, and gene conversion of Ig genes by the deamination of deoxycytidine, followed by error-prone mismatch- or base-excision DNA repair. These processes are crucial for the generation of genetically diverse, high affinity antibody and robust humoral immunity, but exact significant genetic damage and promote cell death. In mice, physiologically significant AID expression was thought to be restricted to antigen-activated, mature B cells in germinal centers. We now demonstrate that low levels of AID in bone marrow immature and transitional B cells suppress the development of autoreactivity. Aicda(-/-) mice exhibit significantly increased serum autoantibody and reduced capacity to purge autoreactive immature and transitional B cells. In vitro, AID deficient immature/transitional B cells are significantly more resistant to anti-IgM-induced apoptosis than their normal counterparts. Thus, early AID expression plays a fundamental and unanticipated role in purging self-reactive immature and transitional B cells during their maturation in the bone marrow.

Authors
Kuraoka, M; Holl, TM; Liao, D; Womble, M; Cain, DW; Reynolds, AE; Kelsoe, G
MLA Citation
Kuraoka, M, Holl, TM, Liao, D, Womble, M, Cain, DW, Reynolds, AE, and Kelsoe, G. "Activation-induced cytidine deaminase mediates central tolerance in B cells." Proc Natl Acad Sci U S A 108.28 (July 12, 2011): 11560-11565.
PMID
21700885
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
108
Issue
28
Publish Date
2011
Start Page
11560
End Page
11565
DOI
10.1073/pnas.1102571108

Role of immune mechanisms in induction of HIV-1 broadly neutralizing antibodies.

Although antibodies can be elicited by HIV-1 infection or immunization, those that are broadly neutralizing (bnAbs) are undetectable in most individuals, and when they do arise in HIV-1 infection, only do so years after transmission. Until recently, the reasons for difficulty in inducing such bnAbs have been obscure. Recent technological advances in isolating bnAbs from rare patients have increased our knowledge of their specificities and features, and along with gene-targeting studies, have also begun uncovering evidence of immunoregulatory roadblocks preventing their induction. One crucial avenue towards developing an effective HIV-1 vaccine is to harness this emerging information into the rational design of immunogens and formulation of adjuvants, such that structural and immunological hurdles to routinely eliciting bnAbs can be overcome.

Authors
Verkoczy, L; Kelsoe, G; Moody, MA; Haynes, BF
MLA Citation
Verkoczy, L, Kelsoe, G, Moody, MA, and Haynes, BF. "Role of immune mechanisms in induction of HIV-1 broadly neutralizing antibodies." Curr Opin Immunol 23.3 (June 2011): 383-390. (Review)
PMID
21524897
Source
pubmed
Published In
Current Opinion in Immunology
Volume
23
Issue
3
Publish Date
2011
Start Page
383
End Page
390
DOI
10.1016/j.coi.2011.04.003

Plexin-D1 is a novel regulator of germinal centers and humoral immune responses.

Long-lived humoral immune responses depend upon the generation of memory B cells and long-lived plasma cells during the germinal center (GC) reaction. These memory compartments, characterized by class-switched IgG and high-affinity Abs, are the basis for successful vaccination. We report that a new member of the plexin family of molecules, plexin-D1, controls the GC reaction and is required for secondary humoral immune responses. Plexin-D1 was not required for B cell maturation, marginal zone precursor development, dark and light zone formation, Igλ(+) and Igκ(+) B cell skewing, B1/B2 development, and the initial extrafollicular response. Plexin-D1 expression was increased following B cell activation, and PlxnD1(-/-) mice exhibited defective GC reactions during T-dependent immune activation. PlxnD1(-/-) B cells showed a defect in migration toward the GC chemokines, CXCL12, CXCL13, and CCL19. Accordingly, PlxnD1(-/-) mice exhibited defective production of IgG1 and IgG2b, but not IgG3 serum Ab, accompanied by reductions in long-lived bone marrow plasmacytes and recall humoral memory responses. These data show a new role for immune plexins in the GC reaction and generation of immunologic memory.

Authors
Holl, EK; O'Connor, BP; Holl, TM; Roney, KE; Zimmermann, AG; Jha, S; Kelsoe, G; Ting, JP-Y
MLA Citation
Holl, EK, O'Connor, BP, Holl, TM, Roney, KE, Zimmermann, AG, Jha, S, Kelsoe, G, and Ting, JP-Y. "Plexin-D1 is a novel regulator of germinal centers and humoral immune responses." J Immunol 186.10 (May 15, 2011): 5603-5611.
PMID
21464091
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
186
Issue
10
Publish Date
2011
Start Page
5603
End Page
5611
DOI
10.4049/jimmunol.1003464

B cell tolerance: putting the horse before the cart.

Authors
Kelsoe, G
MLA Citation
Kelsoe, G. "B cell tolerance: putting the horse before the cart." Arthritis Rheum 63.5 (May 2011): 1173-1176.
PMID
21538309
Source
pubmed
Published In
Arthritis and Rheumatism
Volume
63
Issue
5
Publish Date
2011
Start Page
1173
End Page
1176
DOI
10.1002/art.30162

Impaired social recognition memory in recombination activating gene 1-deficient mice.

The recombination activating genes (RAGs) encode two enzymes that play key roles in the adaptive immune system. RAG1 and RAG2 mediate VDJ recombination, a process necessary for the maturation of B- and T-cells. Interestingly, RAG1 is also expressed in the brain, particularly in areas of high neural density such as the hippocampus, although its function is unknown. We tested evidence that RAG1 plays a role in brain function using a social recognition memory task, an assessment of the acquisition and retention of conspecific identity. In a first experiment, we found that RAG1-deficient mice show impaired social recognition memory compared to mice wildtype for the RAG1 allele. In a second experiment, by breeding to homogenize background genotype, we found that RAG1-deficient mice show impaired social recognition memory relative to heterozygous or RAG2-deficient littermates. Because RAG1 and RAG2 null mice are both immunodeficient, the results suggest that the memory impairment is not an indirect effect of immunological dysfunction. RAG1-deficient mice show normal habituation to non-socially derived odors and habituation to an open-field, indicating that the observed effect is not likely a result of a general deficit in habituation to novelty. These data trace the origin of the impairment in social recognition memory in RAG1-deficient mice to the RAG1 gene locus and implicate RAG1 in memory formation.

Authors
McGowan, PO; Hope, TA; Meck, WH; Kelsoe, G; Williams, CL
MLA Citation
McGowan, PO, Hope, TA, Meck, WH, Kelsoe, G, and Williams, CL. "Impaired social recognition memory in recombination activating gene 1-deficient mice." Brain Res 1383 (April 6, 2011): 187-195.
PMID
21354115
Source
pubmed
Published In
Brain Research
Volume
1383
Publish Date
2011
Start Page
187
End Page
195
DOI
10.1016/j.brainres.2011.02.054

AID expression during B-cell development: searching for answers.

Expression of activation-induced cytidine deaminase (AID) by germinal center (GC) B cells drives the processes of immunoglobulin (Ig) somatic hypermutation (SHM) and class switch recombination (CSR) necessary for the generation of high affinity IgG serum antibody and the memory B-cell compartment. Increasing evidence indicates that AID is also expressed at low levels in developing B cells but to date, this early, developmentally regulated AID expression has no known function. Does the timing and extent of AID expression in developmentally immature, non-GC B cells provide clues to reveal its physiologic role?

Authors
Kuraoka, M; McWilliams, L; Kelsoe, G
MLA Citation
Kuraoka, M, McWilliams, L, and Kelsoe, G. "AID expression during B-cell development: searching for answers." Immunol Res 49.1-3 (April 2011): 3-13.
PMID
21136202
Source
pubmed
Published In
Immunologic Research
Volume
49
Issue
1-3
Publish Date
2011
Start Page
3
End Page
13
DOI
10.1007/s12026-010-8185-7

A novel pathway for self-tolerance: AID and Somatic Hypermutation in the Human Fetus

Authors
McWilliams, LM; Kuraoka, M; Kelsoe, GH
MLA Citation
McWilliams, LM, Kuraoka, M, and Kelsoe, GH. "A novel pathway for self-tolerance: AID and Somatic Hypermutation in the Human Fetus." February 2011.
Source
wos-lite
Published In
Journal of Allergy and Clinical Immunology
Volume
127
Issue
2
Publish Date
2011
Start Page
AB91
End Page
AB91
DOI
10.1016/j.jaci.2010.12.369

Inflammation triggers emergency granulopoiesis through a density-dependent feedback mechanism.

Normally, neutrophil pools are maintained by homeostatic mechanisms that require the transcription factor C/EBPα. Inflammation, however, induces neutrophilia through a distinct pathway of "emergency" granulopoiesis that is dependent on C/EBPβ. Here, we show in mice that alum triggers emergency granulopoiesis through the IL-1RI-dependent induction of G-CSF. G-CSF/G-CSF-R neutralization impairs proliferative responses of hematopoietic stem and progenitor cells (HSPC) to alum, but also abrogates the acute mobilization of BM neutrophils, raising the possibility that HSPC responses to inflammation are an indirect result of the exhaustion of BM neutrophil stores. The induction of neutropenia, via depletion with Gr-1 mAb or myeloid-specific ablation of Mcl-1, elicits G-CSF via an IL-1RI-independent pathway, stimulating granulopoietic responses indistinguishable from those induced by adjuvant. Notably, C/EBPβ, thought to be necessary for enhanced generative capacity of BM, is dispensable for increased proliferation of HSPC to alum or neutropenia, but plays a role in terminal neutrophil differentiation during granulopoietic recovery. We conclude that alum elicits a transient increase in G-CSF production via IL-1RI for the mobilization of BM neutrophils, but density-dependent feedback sustains G-CSF for accelerated granulopoiesis.

Authors
Cain, DW; Snowden, PB; Sempowski, GD; Kelsoe, G
MLA Citation
Cain, DW, Snowden, PB, Sempowski, GD, and Kelsoe, G. "Inflammation triggers emergency granulopoiesis through a density-dependent feedback mechanism." PLoS One 6.5 (2011): e19957-.
Website
http://hdl.handle.net/10161/10903
PMID
21655273
Source
pubmed
Published In
PloS one
Volume
6
Issue
5
Publish Date
2011
Start Page
e19957
DOI
10.1371/journal.pone.0019957

H3N2 influenza infection elicits more cross-reactive and less clonally expanded anti-hemagglutinin antibodies than influenza vaccination.

BACKGROUND: During the recent H1N1 influenza pandemic, excess morbidity and mortality was seen in young but not older adults suggesting that prior infection with influenza strains may have protected older subjects. In contrast, a history of recent seasonal trivalent vaccine in younger adults was not associated with protection. METHODS AND FINDINGS: To study hemagglutinin (HA) antibody responses in influenza immunization and infection, we have studied the day 7 plasma cell repertoires of subjects immunized with seasonal trivalent inactivated influenza vaccine (TIV) and compared them to the plasma cell repertoires of subjects experimentally infected (EI) with influenza H3N2 A/Wisconsin/67/2005. The majority of circulating plasma cells after TIV produced influenza-specific antibodies, while most plasma cells after EI produced antibodies that did not react with influenza HA. While anti-HA antibodies from TIV subjects were primarily reactive with single or few HA strains, anti-HA antibodies from EI subjects were isolated that reacted with multiple HA strains. Plasma cell-derived anti-HA antibodies from TIV subjects showed more evidence of clonal expansion compared with antibodies from EI subjects. From an H3N2-infected subject, we isolated a 4-member clonal lineage of broadly cross-reactive antibodies that bound to multiple HA subtypes and neutralized both H1N1 and H3N2 viruses. This broad reactivity was not detected in post-infection plasma suggesting this broadly reactive clonal lineage was not immunodominant in this subject. CONCLUSION: The presence of broadly reactive subdominant antibody responses in some EI subjects suggests that improved vaccine designs that make broadly reactive antibody responses immunodominant could protect against novel influenza strains.

Authors
Moody, MA; Zhang, R; Walter, EB; Woods, CW; Ginsburg, GS; McClain, MT; Denny, TN; Chen, X; Munshaw, S; Marshall, DJ; Whitesides, JF; Drinker, MS; Amos, JD; Gurley, TC; Eudailey, JA; Foulger, A; DeRosa, KR; Parks, R; Meyerhoff, RR; Yu, J-S; Kozink, DM; Barefoot, BE; Ramsburg, EA; Khurana, S; Golding, H; Vandergrift, NA; Alam, SM; Tomaras, GD; Kepler, TB; Kelsoe, G; Liao, H-X; Haynes, BF
MLA Citation
Moody, MA, Zhang, R, Walter, EB, Woods, CW, Ginsburg, GS, McClain, MT, Denny, TN, Chen, X, Munshaw, S, Marshall, DJ, Whitesides, JF, Drinker, MS, Amos, JD, Gurley, TC, Eudailey, JA, Foulger, A, DeRosa, KR, Parks, R, Meyerhoff, RR, Yu, J-S, Kozink, DM, Barefoot, BE, Ramsburg, EA, Khurana, S, Golding, H, Vandergrift, NA, Alam, SM, Tomaras, GD, Kepler, TB, Kelsoe, G, Liao, H-X, and Haynes, BF. "H3N2 influenza infection elicits more cross-reactive and less clonally expanded anti-hemagglutinin antibodies than influenza vaccination." PLoS One 6.10 (2011): e25797-.
PMID
22039424
Source
pubmed
Published In
PloS one
Volume
6
Issue
10
Publish Date
2011
Start Page
e25797
DOI
10.1371/journal.pone.0025797

A new guise for hyper-IgM syndrome.

Authors
Kelsoe, G; Cain, D
MLA Citation
Kelsoe, G, and Cain, D. "A new guise for hyper-IgM syndrome." Blood 116.26 (December 23, 2010): 5785-5786.
PMID
21183692
Source
pubmed
Published In
Blood
Volume
116
Issue
26
Publish Date
2010
Start Page
5785
End Page
5786
DOI
10.1182/blood-2010-09-303404

Crystal structure of a non-neutralizing antibody to the HIV-1 gp41 membrane-proximal external region.

The monoclonal antibody 13H11 shares part of its epitope in the HIV-1 gp41 membrane-proximal external region (MPER) with the rare, broadly neutralizing human antibody 2F5. Although 13H11 partially cross-blocked 2F5 binding, 13H11 is non-neutralizing and does not block 2F5 neutralization. We show that unlike 2F5, 13H11 binds to a well-defined helical MPER structure that is consistent with the structure of gp41 in a post-fusion six-helix bundle conformation.

Authors
Nicely, NI; Dennison, SM; Spicer, L; Scearce, RM; Kelsoe, G; Ueda, Y; Chen, H; Liao, H-X; Alam, SM; Haynes, BF
MLA Citation
Nicely, NI, Dennison, SM, Spicer, L, Scearce, RM, Kelsoe, G, Ueda, Y, Chen, H, Liao, H-X, Alam, SM, and Haynes, BF. "Crystal structure of a non-neutralizing antibody to the HIV-1 gp41 membrane-proximal external region." Nat Struct Mol Biol 17.12 (December 2010): 1492-1494.
PMID
21076400
Source
pubmed
Published In
Nature Structural & Molecular Biology
Volume
17
Issue
12
Publish Date
2010
Start Page
1492
End Page
1494
DOI
10.1038/nsmb.1944

Distinct granuloma responses in C57BL/6J and BALB/cByJ mice in response to pristane.

Granuloma formation is an inflammatory response of the host against invading pathogens or indigestible substances. We generated mesenteric oil granulomas by injecting pristane into the peritoneal cavity (PC) of mice, and compared oil granuloma formation in the C57BL/6J and BALB/cByJ strains of mice. The formation and kinetics of oil granulomas were distinct between the two strains. In C57BL/6J mice, injected pristane induced oil granuloma formation at both the mesenteric centers (MG) and margins (SG). MG was resolving by 11 weeks, and SG persisted. In BALB/cByJ mice, MG developed slower but persisted longer than in C57BL/6J mice, and SG resolved sooner than in C57BL/6J mice. Injection of India ink revealed that phagocytes were localised mainly to the SG in C57BL/6J mice, but were located diffusely in both MG and SG of BALB/cByJ mice. SG cells expressed more monocyte chemotactic protein-1 (MCP-1) mRNA than MG cells in C57BL/6J mice, but there was no difference in MCP-1 expression between the MG and SG in BALB/cByJ mice. These observations suggest that the recruitment of inflammatory leucocytes under the direction of chemokines differentiates the patterns of granuloma responses to pristane in C57BL/6J and BALB/cByJ mice.

Authors
Chen, H; Liao, D; Cain, D; McLeod, I; Ueda, Y; Guan, Z; Raetz, C; Kelsoe, G
MLA Citation
Chen, H, Liao, D, Cain, D, McLeod, I, Ueda, Y, Guan, Z, Raetz, C, and Kelsoe, G. "Distinct granuloma responses in C57BL/6J and BALB/cByJ mice in response to pristane." Int J Exp Pathol 91.5 (October 2010): 460-471.
PMID
20681981
Source
pubmed
Published In
International Journal of Experimental Pathology
Volume
91
Issue
5
Publish Date
2010
Start Page
460
End Page
471
DOI
10.1111/j.1365-2613.2010.00725.x

Genetic regulation of pristane-induced oil granuloma responses.

Oil granuloma (OG) induced by intraperitoneal injection of pristane represents a non-infectious granuloma. Oil granuloma has been characterized, but the regulation of its formation still remains unknown. To address this, we injected pristane into various mice deficient for genes including, linker for activation of T cells (LAT), μMT, LTα, TNFα, IL-6. T cell deficient mice (LAT(-/-) ) responded to pristane by developing serosal granuloma and mesenteric granuloma (MG) as in wild type mice. The absence of B cells blocked serosal granuloma (SG) formation and diminished MG development in response to pristane. However, even when a comparable number of B cells were present in the mesentery, the absence of TNFα resulted in similar defects in OG formation after pristane treatment, demonstrating that both B cells and TNFα are very crucial for pristane-induced OG formation. Interestingly, IL-6(-/-) mice had intact MG formation; however, SG organization was impaired. These studies provide insight into granulomateous pathology induced by non-infectious substances for example, biomedical sutures.

Authors
Chen, H; Liao, D; Holl, TM; Snowden, P; Ueda, Y; Kelsoe, G
MLA Citation
Chen, H, Liao, D, Holl, TM, Snowden, P, Ueda, Y, and Kelsoe, G. "Genetic regulation of pristane-induced oil granuloma responses." Int J Exp Pathol 91.5 (October 2010): 472-483.
PMID
20804539
Source
pubmed
Published In
International Journal of Experimental Pathology
Volume
91
Issue
5
Publish Date
2010
Start Page
472
End Page
483
DOI
10.1111/j.1365-2613.2010.00732.x

Stromal cell independent B cell development in vitro: generation and recovery of autoreactive clones.

We describe and characterize a stromal cell independent culture system that efficiently supports pro-B cell to IgM+ B cell development with near normal levels of IgH and Igkappa diversity. Pro-B cells present in non-adherent bone marrow cells proliferate in the presence of IL-7 and subsequent to the removal of IL-7 and addition of BAFF, differentiate normally into IgM+ B cells. B cell development in vitro closely follows the patterns of development in vivo with culture-derived (CD) B cells demonstrating characteristic patterns of surface antigen expression and gene activation. IgM+ CD B cells respond to TLR stimulation by proliferation and differentiation into antibody-secreting cells. Self-reactive IgM+ B cell development is blocked in 3H9 IgH knockin mice; however, cultures of 3H9 IgH knockin pro-B cells yields high frequencies of "forbidden", autoreactive IgM+ B cells. Furthermore, serum IgG autoantibody exceeded that present in autoimmune, C4(-/-) animals following the reconstitution of RAG1(-/-) mice with IgM+ CD cells derived from BL/6 mice.

Authors
Holl, TM; Haynes, BF; Kelsoe, G
MLA Citation
Holl, TM, Haynes, BF, and Kelsoe, G. "Stromal cell independent B cell development in vitro: generation and recovery of autoreactive clones." J Immunol Methods 354.1-2 (March 31, 2010): 53-67.
PMID
20109461
Source
pubmed
Published In
Journal of Immunological Methods
Volume
354
Issue
1-2
Publish Date
2010
Start Page
53
End Page
67
DOI
10.1016/j.jim.2010.01.007

Autoreactivity in an HIV-1 broadly reactive neutralizing antibody variable region heavy chain induces immunologic tolerance.

We previously reported that some of the rare broadly reactive, HIV-1 neutralizing antibodies are polyreactive, leading to the hypothesis that induction of these types of neutralizing antibody may be limited by immunologic tolerance. However, the notion that such antibodies are sufficiently autoreactive to trigger B cell tolerance is controversial. To test directly whether rare neutralizing HIV-1 antibodies can activate immunologic tolerance mechanisms, we generated a knock-in mouse in which the Ig heavy chain (HC) variable region rearrangement (V(H)DJ(H)) from the polyreactive and broadly neutralizing human monoclonal antibody 2F5 was targeted into the mouse Igh locus. In vitro, this insertion resulted in chimeric human/mouse 2F5 antibodies that were functionally similar to the human 2F5 antibody, including comparable reactivity to human and murine self-antigens. In vivo, the 2F5 V(H)DJ(H) insertion supported development of large- and small pre-B cells that expressed the chimeric human/mouse Igmu chain but not the production of immature B cells expressing membrane IgM. The developmental arrest exhibited in 2F5 V(H)DJ(H) knock-in mice is characteristic of other knock-in strains that express the Ig HC variable region of autoreactive antibodies and is consistent with the loss of immature B cells bearing 2F5 chimeric antibodies to central tolerance mechanisms. Moreover, homozygous 2F5 V(H)DJ(H) knock-in mice support reduced numbers of residual splenic B cells with low surface IgM density, severely diminished serum IgM levels, but normal to elevated quantities of serum IgGs that did not react with autoantigens. These features are consistent with elimination of 2F5 HC autoreactivity by additional negative selection mechanism(s) in the periphery.

Authors
Verkoczy, L; Diaz, M; Holl, TM; Ouyang, Y-B; Bouton-Verville, H; Alam, SM; Liao, H-X; Kelsoe, G; Haynes, BF
MLA Citation
Verkoczy, L, Diaz, M, Holl, TM, Ouyang, Y-B, Bouton-Verville, H, Alam, SM, Liao, H-X, Kelsoe, G, and Haynes, BF. "Autoreactivity in an HIV-1 broadly reactive neutralizing antibody variable region heavy chain induces immunologic tolerance." Proc Natl Acad Sci U S A 107.1 (January 5, 2010): 181-186.
PMID
20018688
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
107
Issue
1
Publish Date
2010
Start Page
181
End Page
186
DOI
10.1073/pnas.0912914107

Functional, non-clonal IgMa-restricted B cell receptor interactions with the HIV-1 envelope gp41 membrane proximal external region.

The membrane proximal external region (MPER) of HIV-1 gp41 has several features that make it an attractive antibody-based vaccine target, but eliciting an effective gp41 MPER-specific protective antibody response remains elusive. One fundamental issue is whether the failure to make gp41 MPER-specific broadly neutralizing antibodies like 2F5 and 4E10 is due to structural constraints with the gp41 MPER, or alternatively, if gp41 MPER epitope-specific B cells are lost to immunological tolerance. An equally important question is how B cells interact with, and respond to, the gp41 MPER epitope, including whether they engage this epitope in a non-canonical manner i.e., by non-paratopic recognition via B cell receptors (BCR). To begin understanding how B cells engage the gp41 MPER, we characterized B cell-gp41 MPER interactions in BALB/c and C57BL/6 mice. Surprisingly, we found that a significant (approximately 7%) fraction of splenic B cells from BALB/c, but not C57BL/6 mice, bound the gp41 MPER via their BCRs. This strain-specific binding was concentrated in IgM(hi) subsets, including marginal zone and peritoneal B1 B cells, and correlated with enriched fractions (approximately 15%) of gp41 MPER-specific IgM secreted by in vitro-activated splenic B cells. Analysis of Igh(a) (BALB/c) and Igh(b) (C57BL/6) congenic mice demonstrated that gp41 MPER binding was controlled by determinants of the Igh(a) locus. Mapping of MPER gp41 interactions with IgM(a) identified MPER residues distinct from those to which mAb 2F5 binds and demonstrated the requirement of Fc C(H) regions. Importantly, gp41 MPER ligation produced detectable BCR-proximal signaling events, suggesting that interactions between gp41 MPER and IgM(a) determinants may elicit partial B cell activation. These data suggest that low avidity, non-paratopic interactions between the gp41 MPER and membrane Ig on naïve B cells may interfere with or divert bnAb responses.

Authors
Verkoczy, L; Moody, MA; Holl, TM; Bouton-Verville, H; Scearce, RM; Hutchinson, J; Alam, SM; Kelsoe, G; Haynes, BF
MLA Citation
Verkoczy, L, Moody, MA, Holl, TM, Bouton-Verville, H, Scearce, RM, Hutchinson, J, Alam, SM, Kelsoe, G, and Haynes, BF. "Functional, non-clonal IgMa-restricted B cell receptor interactions with the HIV-1 envelope gp41 membrane proximal external region. (Published online)" PLoS One 4.10 (October 6, 2009): e7215-.
PMID
19806186
Source
pubmed
Published In
PloS one
Volume
4
Issue
10
Publish Date
2009
Start Page
e7215
DOI
10.1371/journal.pone.0007215

Activation-induced cytidine deaminase expression and activity in the absence of germinal centers: insights into hyper-IgM syndrome.

Somatic hypermutation normally occurs as a consequence of the expression of activation-induced cytidine deaminase (AID) by Ag-activated, mature B cells during T cell-dependent germinal center responses. Nonetheless, despite their inability to express CD154 and initiate GC responses, patients with type 1 hyper-IgM syndrome (HIGM1) support populations of IgM(+)IgD(+)CD27(+) B cells that express mutated Ig genes. The origin of these mutated B cells is unknown; the IgM(+)IgD(+)CD27(+) cells do not express AID and appear to acquire mutations independent of stringent selection by Ag. Here, we demonstrate that immature/transitional 1 B cells from the bone marrow of CD154-deficient mice express AID and acquire Ig mutations that lack the hallmarks of antigenic selection via BCR signaling. Comparable levels of AID expression was found in developmentally immature B cells recovered from murine fetal liver and from human immature/transitional 1 B cells recovered from umbilical cord blood. AID expression in human fetal liver was also robust, approaching that of human tonsil tissue and the human germinal center B cell line, Ramos. These observations led us to conclude that AID expression in developing human B cells is the origin of the mutated IgM(+)IgD(+)CD27(+) B cells present in HIGM1 patients, and we propose that both mice and humans share a latent, AID-dependent pathway for the preimmune diversification of B lymphocytes that is more prominent in chicken, sheep, and rabbits.

Authors
Kuraoka, M; Liao, D; Yang, K; Allgood, SD; Levesque, MC; Kelsoe, G; Ueda, Y
MLA Citation
Kuraoka, M, Liao, D, Yang, K, Allgood, SD, Levesque, MC, Kelsoe, G, and Ueda, Y. "Activation-induced cytidine deaminase expression and activity in the absence of germinal centers: insights into hyper-IgM syndrome." J Immunol 183.5 (September 1, 2009): 3237-3248.
PMID
19667096
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
183
Issue
5
Publish Date
2009
Start Page
3237
End Page
3248
DOI
10.4049/jimmunol.0901548

HIV-1 envelope induces memory B cell responses that correlate with plasma antibody levels after envelope gp120 protein vaccination or HIV-1 infection.

Successful vaccines (i.e., tetanus and diphtheria) can induce long-lived Ab levels that are maintained by bone marrow plasma cells and plasma Ab levels do not correlate with numbers of blood memory B cells. Destruction of CD4(+) T cells early in HIV-1 acute infection may result in insufficient induction of neutralizing Ab responses; thus, an HIV-1 vaccine should elicit high levels of durable Abs by long-lived plasma cells to be protective. We asked if HIV-1 envelope-specific memory responses were sustained by memory B cells in the settings of HIV-1 gp120 envelope vaccination and chronic HIV-1 infection. Levels of anti-HIV-1 envelope plasma Abs and memory B cells were found to correlate in both settings. Moreover, whereas the expected half-life of plasma Ab levels to protein vaccines was >10 years when maintained by long-lived plasma cells, anti-envelope Ab level half-lives were approximately 33-81 wk in plasma from antiretroviral drug-treated HIV-1(+) subjects. In contrast, anti-p55 Gag Ab level half-life was 648 wk, and Ab titers against influenza did not decay in-between yearly or biennial influenza vaccine boosts in the same patients. These data demonstrated that HIV-1 envelope induces predominantly short-lived memory B cell-dependent plasma Abs in the settings of envelope vaccination and HIV-1 infection. The inability to generate high titers of long-lived anti-envelope Abs is a major hurdle to overcome for the development of a successful HIV-1 vaccine.

Authors
Bonsignori, M; Moody, MA; Parks, RJ; Holl, TM; Kelsoe, G; Hicks, CB; Vandergrift, N; Tomaras, GD; Haynes, BF
MLA Citation
Bonsignori, M, Moody, MA, Parks, RJ, Holl, TM, Kelsoe, G, Hicks, CB, Vandergrift, N, Tomaras, GD, and Haynes, BF. "HIV-1 envelope induces memory B cell responses that correlate with plasma antibody levels after envelope gp120 protein vaccination or HIV-1 infection." J Immunol 183.4 (August 15, 2009): 2708-2717.
PMID
19625640
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
183
Issue
4
Publish Date
2009
Start Page
2708
End Page
2717
DOI
10.4049/jimmunol.0901068

Polyclonal B cell differentiation and loss of gastrointestinal tract germinal centers in the earliest stages of HIV-1 infection.

The antibody response to HIV-1 does not appear in the plasma until approximately 2-5 weeks after transmission, and neutralizing antibodies to autologous HIV-1 generally do not become detectable until 12 weeks or more after transmission. Moreover, levels of HIV-1-specific antibodies decline on antiretroviral treatment. The mechanisms of this delay in the appearance of anti-HIV-1 antibodies and of their subsequent rapid decline are not known. While the effect of HIV-1 on depletion of gut CD4(+) T cells in acute HIV-1 infection is well described, we studied blood and tissue B cells soon after infection to determine the effect of early HIV-1 on these cells.In human participants, we analyzed B cells in blood as early as 17 days after HIV-1 infection, and in terminal ileum inductive and effector microenvironments beginning at 47 days after infection. We found that HIV-1 infection rapidly induced polyclonal activation and terminal differentiation of B cells in blood and in gut-associated lymphoid tissue (GALT) B cells. The specificities of antibodies produced by GALT memory B cells in acute HIV-1 infection (AHI) included not only HIV-1-specific antibodies, but also influenza-specific and autoreactive antibodies, indicating very early onset of HIV-1-induced polyclonal B cell activation. Follicular damage or germinal center loss in terminal ileum Peyer's patches was seen with 88% of follicles exhibiting B or T cell apoptosis and follicular lysis.Early induction of polyclonal B cell differentiation, coupled with follicular damage and germinal center loss soon after HIV-1 infection, may explain both the high rate of decline in HIV-1-induced antibody responses and the delay in plasma antibody responses to HIV-1. Please see later in the article for Editors' Summary.

Authors
Levesque, MC; Moody, MA; Hwang, K-K; Marshall, DJ; Whitesides, JF; Amos, JD; Gurley, TC; Allgood, S; Haynes, BB; Vandergrift, NA; Plonk, S; Parker, DC; Cohen, MS; Tomaras, GD; Goepfert, PA; Shaw, GM; Schmitz, JE; Eron, JJ; Shaheen, NJ; Hicks, CB; Liao, H-X; Markowitz, M; Kelsoe, G; Margolis, DM; Haynes, BF
MLA Citation
Levesque, MC, Moody, MA, Hwang, K-K, Marshall, DJ, Whitesides, JF, Amos, JD, Gurley, TC, Allgood, S, Haynes, BB, Vandergrift, NA, Plonk, S, Parker, DC, Cohen, MS, Tomaras, GD, Goepfert, PA, Shaw, GM, Schmitz, JE, Eron, JJ, Shaheen, NJ, Hicks, CB, Liao, H-X, Markowitz, M, Kelsoe, G, Margolis, DM, and Haynes, BF. "Polyclonal B cell differentiation and loss of gastrointestinal tract germinal centers in the earliest stages of HIV-1 infection." PLoS medicine 6.7 (July 7, 2009): e1000107-.
Website
http://hdl.handle.net/10161/10895
PMID
19582166
Source
epmc
Published In
PLoS medicine
Volume
6
Issue
7
Publish Date
2009
Start Page
e1000107
DOI
10.1371/journal.pmed.1000107

Conserved cryptic recombination signals in Vkappa gene segments are cleaved in small pre-B cells.

BACKGROUND: The cleavage of recombination signals (RS) at the boundaries of immunoglobulin V, D, and J gene segments initiates the somatic generation of the antigen receptor genes expressed by B lymphocytes. RS contain a conserved heptamer and nonamer motif separated by non-conserved spacers of 12 or 23 nucleotides. Under physiologic conditions, V(D)J recombination follows the "12/23 rule" to assemble functional antigen-receptor genes, i.e., cleavage and recombination occur only between RS with dissimilar spacer types. Functional, cryptic RS (cRS) have been identified in VH gene segments; these VH cRS were hypothesized to facilitate self-tolerance by mediating VH --> VHDJH replacements. At the Igkappa locus, however, secondary, de novo rearrangements can delete autoreactive VkappaJkappa joins. Thus, under the hypothesis that V-embedded cRS are conserved to facilitate self-tolerance by mediating V-replacement rearrangements, there would be little selection for Vkappa cRS. Recent studies have demonstrated that VH cRS cleavage is only modestly more efficient than V(D)J recombination in violation of the 12/23 rule and first occurs in pro-B cells unable to interact with exogenous antigens. These results are inconsistent with a model of cRS cleavage during autoreactivity-induced VH gene replacement. RESULTS: To test the hypothesis that cRS are absent from Vkappa gene segments, a corollary of the hypothesis that the need for tolerizing VH replacements is responsible for the selection pressure to maintain VH cRS, we searched for cRS in mouse Vkappa gene segments using a statistical model of RS. Scans of 135 mouse Vkappa gene segments revealed highly conserved cRS that were shown to be cleaved in the 103/BCL2 cell line and mouse bone marrow B cells. Analogous to results for VH cRS, we find that Vkappa cRS are conserved at multiple locations in Vkappa gene segments and are cleaved in pre-B cells. CONCLUSION: Our results, together with those for VH cRS, support a model of cRS cleavage in which cleavage is independent of BCR-specificity. Our results are inconsistent with the hypothesis that cRS are conserved solely to support receptor editing. The extent to which these sequences are conserved, and their pattern of conservation, suggest that they may serve an as yet unidentified purpose.

Authors
Lieberman, AE; Kuraoka, M; Davila, M; Kelsoe, G; Cowell, LG
MLA Citation
Lieberman, AE, Kuraoka, M, Davila, M, Kelsoe, G, and Cowell, LG. "Conserved cryptic recombination signals in Vkappa gene segments are cleaved in small pre-B cells. (Published online)" BMC Immunol 10 (June 25, 2009): 37-.
PMID
19555491
Source
pubmed
Published In
BMC Immunology
Volume
10
Publish Date
2009
Start Page
37
DOI
10.1186/1471-2172-10-37

IL-1R type I-dependent hemopoietic stem cell proliferation is necessary for inflammatory granulopoiesis and reactive neutrophilia.

Infections and inflammation trigger neutrophilias that are supported by a hematopoietic program of accelerated granulopoiesis known as emergency granulopoiesis. The intrinsic factors that drive reactive neutrophilias and emergency granulopoiesis have been inferred but not demonstrated. Here, we show that alum cannot elicit reactive neutrophilias in IL-1R type I (IL-1RI)(-/-) mice, whereas other inflammatory responses, including eosinophilia and Ab production, remain intact. Analysis of this specific impairment revealed an unanticipated role for IL-1RI in supporting increased proliferation by granulocyte/macrophage progenitors and, surprisingly, multipotent progenitors and hematopoietic stem cells (HSC). Indeed, HSC and multipotent progenitor proliferative responses were most suppressed in IL-1RI(-/-) mice, suggesting a critical role for their proliferation in inflammatory granulopoiesis. Whereas IL-1 drives increased HSC proliferation directly in vitro, IL-1RI expression by radiation-resistant host cells was both necessary and sufficient for alum-induced HSC, multipotent progenitor, and granulocyte/macrophage progenitor proliferation and reactive neutrophilias in radiation chimeric mice. Thus, IL-1 plays a necessary, but indirect, role in the support of alum-induced neutrophilias by expanding both pluripotent and myeloid progenitor compartments to accelerate granulopoiesis.

Authors
Ueda, Y; Cain, DW; Kuraoka, M; Kondo, M; Kelsoe, G
MLA Citation
Ueda, Y, Cain, DW, Kuraoka, M, Kondo, M, and Kelsoe, G. "IL-1R type I-dependent hemopoietic stem cell proliferation is necessary for inflammatory granulopoiesis and reactive neutrophilia." J Immunol 182.10 (May 15, 2009): 6477-6484.
PMID
19414802
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
182
Issue
10
Publish Date
2009
Start Page
6477
End Page
6484
DOI
10.4049/jimmunol.0803961

Effects of acute and chronic inflammation on B-cell development and differentiation.

Recently, our understanding of hematopoiesis and the development of the immune system has fundamentally changed, leading to significant discoveries with important clinical relevance. Hematopoiesis, once described in terms of irreversible and discrete developmental branch points, is now understood to exist as a collection of alternative developmental pathways capable of generating functionally identical progeny. Developmental commitment to a particular blood-cell lineage is gradually acquired and reflects both cell intrinsic and extrinsic signals. Chief among the extrinsic factors are the environmental cues of hematopoietic microenvironments that comprise specific "developmental niches" that support hematopoietic stem and progenitor cells. Most of this new understanding comes from the study of normal, steady-state hematopoiesis, but there is ample reason to expect that special developmental and/or differentiative mechanisms operate in response to inflammation. For example, both stem and progenitor cells are now known to express Toll-like receptors that can influence hematopoietic cell fates in response to microbial products. Likewise, proinflammatory cytokines mobilize hematopoietic stem cells to peripheral tissues. In this Perspective, we review inflammation's effects on central and extramedullary B lymphopoiesis and discuss the potential consequences of peripheral B-cell development in the context of systemic autoimmune diseases.

Authors
Cain, D; Kondo, M; Chen, H; Kelsoe, G
MLA Citation
Cain, D, Kondo, M, Chen, H, and Kelsoe, G. "Effects of acute and chronic inflammation on B-cell development and differentiation." J Invest Dermatol 129.2 (February 2009): 266-277. (Review)
PMID
19148216
Source
pubmed
Published In
Journal of Investigative Dermatology
Volume
129
Issue
2
Publish Date
2009
Start Page
266
End Page
277
DOI
10.1038/jid.2008.286

A large-scale analysis of immunoglobulin sequences derived from plasmablasts/plasma cells in acute HIV-1 infection subjects

Authors
Munshaw, S; Liao, H; Dixon, A; Chen, X; Nagel, A; Derosa, K; Parks, R; Amos, J; Whitesides, JF; Marshalls, DJ; Yang, Y; Gao, F; Tomaras, GD; Moody, MA; Kelsoe, GH; Shea, TC; Margolis, DM; Markowitz, M; Goepfert, P; Shaw, G; Haynes, BF; Kepler, TB
MLA Citation
Munshaw, S, Liao, H, Dixon, A, Chen, X, Nagel, A, Derosa, K, Parks, R, Amos, J, Whitesides, JF, Marshalls, DJ, Yang, Y, Gao, F, Tomaras, GD, Moody, MA, Kelsoe, GH, Shea, TC, Margolis, DM, Markowitz, M, Goepfert, P, Shaw, G, Haynes, BF, and Kepler, TB. "A large-scale analysis of immunoglobulin sequences derived from plasmablasts/plasma cells in acute HIV-1 infection subjects." 2009.
Source
wos-lite
Published In
Retrovirology
Volume
6
Publish Date
2009

Generation of antibody responses to HIV-1 membrane proximal external region (MPER) antigen

Authors
Holl, TM; Kuraoka, M; Liao, D; Verkoczy, L; Moody, MA; Alam, M; Liao, H; Haynes, BF; Kelsoe, GH
MLA Citation
Holl, TM, Kuraoka, M, Liao, D, Verkoczy, L, Moody, MA, Alam, M, Liao, H, Haynes, BF, and Kelsoe, GH. "Generation of antibody responses to HIV-1 membrane proximal external region (MPER) antigen." RETROVIROLOGY 6 (2009).
Source
wos-lite
Published In
Retrovirology
Volume
6
Publish Date
2009

Maintenance of long-lived plasma cells and serological memory despite mature and memory B cell depletion during CD20 immunotherapy in mice.

CD20 mAb-mediated B cell depletion is an effective treatment for B cell malignancies and some autoimmune diseases. However, the full effects of B cell depletion on natural, primary, and secondary Ab responses and the maintenance of Ag-specific serum Ig levels are largely unknown. The relationship between memory B cells, long-lived plasma cells, and long-lived humoral immunity also remains controversial. To address the roles of B cell subsets in the longevity of humoral responses, mature B cells were depleted in mice using CD20 mAb. Peritoneal B cell depletion reduced natural and Ag-induced IgM responses. Otherwise, CD20+ B cell depletion prevented humoral immune responses and class switching and depleted existing and adoptively transferred B cell memory. Nonetheless, B cell depletion did not affect serum Ig levels, Ag-specific Ab titers, or bone marrow Ab-secreting plasma cell numbers. Coblockade of LFA-1 and VLA-4 adhesion molecules temporarily depleted long-lived plasma cells from the bone marrow. CD20+ B cell depletion plus LFA-1/VLA-4 mAb treatment significantly prolonged Ag-specific plasma cell depletion from the bone marrow, with a significant decrease in Ag-specific serum IgG. Collectively, these results support previous claims that bone marrow plasma cells are intrinsically long-lived. Furthermore, these studies now demonstrate that mature and memory B cells are not required for maintaining bone marrow plasma cell numbers, but are required for repopulation of plasma cell-deficient bone marrow. Thereby, depleting mature and memory B cells does not have a dramatic negative effect on preexisting Ab levels.

Authors
DiLillo, DJ; Hamaguchi, Y; Ueda, Y; Yang, K; Uchida, J; Haas, KM; Kelsoe, G; Tedder, TF
MLA Citation
DiLillo, DJ, Hamaguchi, Y, Ueda, Y, Yang, K, Uchida, J, Haas, KM, Kelsoe, G, and Tedder, TF. "Maintenance of long-lived plasma cells and serological memory despite mature and memory B cell depletion during CD20 immunotherapy in mice." J Immunol 180.1 (January 1, 2008): 361-371.
PMID
18097037
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
180
Issue
1
Publish Date
2008
Start Page
361
End Page
371

Multiple, conserved cryptic recombination signals in VH gene segments: detection of cleavage products only in pro B cells.

Receptor editing is believed to play the major role in purging newly formed B cell compartments of autoreactivity by the induction of secondary V(D)J rearrangements. In the process of immunoglobulin heavy (H) chain editing, these secondary rearrangements are mediated by direct V(H)-to-J(H) joining or cryptic recombination signals (cRSs) within V(H) gene segments. Using a statistical model of RS, we have identified potential cRSs within V(H) gene segments at conserved sites flanking complementarity-determining regions 1 and 2. These cRSs are active in extrachromosomal recombination assays and cleaved during normal B cell development. Cleavage of multiple V(H) cRSs was observed in the bone marrow of C57BL/6 and RAG2:GFP and microMT congenic animals, and we determined that cRS cleavage efficiencies are 30-50-fold lower than a physiological RS. cRS signal ends are abundant in pro-B cells, including those recovered from microMT mice, but undetectable in pre- or immature B cells. Thus, V(H) cRS cleavage regularly occurs before the generation of functional preBCR and BCR. Conservation of cRSs distal from the 3' end of V(H) gene segments suggests a function for these cryptic signals other than V(H) gene replacement.

Authors
Davila, M; Liu, F; Cowell, LG; Lieberman, AE; Heikamp, E; Patel, A; Kelsoe, G
MLA Citation
Davila, M, Liu, F, Cowell, LG, Lieberman, AE, Heikamp, E, Patel, A, and Kelsoe, G. "Multiple, conserved cryptic recombination signals in VH gene segments: detection of cleavage products only in pro B cells." J Exp Med 204.13 (December 24, 2007): 3195-3208.
Website
http://hdl.handle.net/10161/10905
PMID
18056287
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
204
Issue
13
Publish Date
2007
Start Page
3195
End Page
3208
DOI
10.1084/jem.20071224

The role of antibody polyspecificity and lipid reactivity in binding of broadly neutralizing anti-HIV-1 envelope human monoclonal antibodies 2F5 and 4E10 to glycoprotein 41 membrane proximal envelope epitopes.

Two neutralizing human mAbs, 2F5 and 4E10, that react with the HIV-1 envelope gp41 membrane proximal region are also polyspecific autoantibodies that bind to anionic phospholipids. To determine the autoantibody nature of these Abs, we have compared their reactivities with human anti-cardiolipin mAbs derived from a primary antiphospholipid syndrome patient. To define the role of lipid polyreactivity in binding of 2F5 and 4E10 mAbs to HIV-1 envelope membrane proximal epitopes, we determined the kinetics of binding of mAbs 2F5 and 4E10 to their nominal gp41 epitopes vs liposome-gp41 peptide conjugates. Both anti-HIV-1 mAbs 2F5 and 4E10 bound to cardiolipin with K(d) values similar to those of autoimmune anti-cardiolipin Abs, IS4 and IS6. Binding kinetics studies revealed that mAb 2F5 and 4E10 binding to their respective gp41 peptide-lipid conjugates could best be defined by a two-step (encounter-docking) conformational change model. In contrast, binding of 2F5 and 4E10 mAbs to linear peptide epitopes followed a simple Langmuir model. A mouse mAb, 13H11, that cross-blocks mAb 2F5 binding to the gp41 epitope did not cross-react with lipids nor did it neutralize HIV-1 viruses. Taken together, these data demonstrate the similarity of 2F5 and 4E10 mAbs to known anti-cardiolipin Abs and support the model that mAb 2F5 and 4E10 binding to HIV-1 involves both viral lipid membrane and gp41 membrane proximal epitopes.

Authors
Alam, SM; McAdams, M; Boren, D; Rak, M; Scearce, RM; Gao, F; Camacho, ZT; Gewirth, D; Kelsoe, G; Chen, P; Haynes, BF
MLA Citation
Alam, SM, McAdams, M, Boren, D, Rak, M, Scearce, RM, Gao, F, Camacho, ZT, Gewirth, D, Kelsoe, G, Chen, P, and Haynes, BF. "The role of antibody polyspecificity and lipid reactivity in binding of broadly neutralizing anti-HIV-1 envelope human monoclonal antibodies 2F5 and 4E10 to glycoprotein 41 membrane proximal envelope epitopes." J Immunol 178.7 (April 1, 2007): 4424-4435.
PMID
17372000
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
178
Issue
7
Publish Date
2007
Start Page
4424
End Page
4435

T-independent activation-induced cytidine deaminase expression, class-switch recombination, and antibody production by immature/transitional 1 B cells.

Inflammation elicits a splenic lymphopoiesis of unknown physiologic significance but one that juxtaposes developing B cells and exogenous Ag. We show that immature and transitional 1 (immature/T1) B cells constitutively express activation-induced cytidine deaminase and B lymphocyte-induced maturation protein 1 in amounts that support accelerated plasmacytic differentiation and limited class-switch recombination. In vivo, activation of immature/T1 B cells by TLR ligands or bacterial vaccine rapidly induces T1 cells to divide, proliferate, and secrete IgM, IgG, or IgA Ab; in vitro, proliferation and differentiation are substantially enhanced by B cell-activating factor. We propose that inflammation-induced extramedullary lymphopoiesis represents a specialized mechanism for innate Ab responses to microbial pathogens.

Authors
Ueda, Y; Liao, D; Yang, K; Patel, A; Kelsoe, G
MLA Citation
Ueda, Y, Liao, D, Yang, K, Patel, A, and Kelsoe, G. "T-independent activation-induced cytidine deaminase expression, class-switch recombination, and antibody production by immature/transitional 1 B cells." J Immunol 178.6 (March 15, 2007): 3593-3601.
PMID
17339456
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
178
Issue
6
Publish Date
2007
Start Page
3593
End Page
3601

Progressive immunoglobulin gene mutations in chronic lymphocytic leukemia: evidence for antigen-driven intraclonal diversification.

Somatic mutations of immunoglobulin genes characterize mature memory B cells, and intraclonal B-cell diversification is typically associated with expansion of B-cell clones with greater affinity for antigen (antigen drive). Evidence for a role of antigen in progression of intraclonal chronic lymphocytic leukemia (CLL) cell diversification in patients with mutated immunoglobulin genes has not been previously presented. We performed a single-cell analysis of immunoglobulin heavy and light chains in 6 patients with somatically mutated CLL-cell immunoglobulin genes and identified 2 patients with multiple related (oligoclonal) subgroups of CLL cells. We constructed genealogic trees of these oligoclonal CLL-cell subgroups and assessed the effects of immunoglobulin somatic mutations on the ratios of replacement and silent amino acid changes in the framework and antigen-binding regions (CDRs) of the immunoglobulin heavy and light chains from each oligoclonal CLL-cell population. In one subject, the amino acid changes were consistent with an antigen-driven progression of clonally related CLL-cell populations. In the other subject, intraclonal diversification was associated with immunoglobulin amino acid changes that would have likely lessened antigen affinity. Taken together, these studies support the hypothesis that in some CLL cases intraclonal diversification is dependent on antigen interactions with immunoglobulin receptors.

Authors
Volkheimer, AD; Weinberg, JB; Beasley, BE; Whitesides, JF; Gockerman, JP; Moore, JO; Kelsoe, G; Goodman, BK; Levesque, MC
MLA Citation
Volkheimer, AD, Weinberg, JB, Beasley, BE, Whitesides, JF, Gockerman, JP, Moore, JO, Kelsoe, G, Goodman, BK, and Levesque, MC. "Progressive immunoglobulin gene mutations in chronic lymphocytic leukemia: evidence for antigen-driven intraclonal diversification." Blood 109.4 (February 15, 2007): 1559-1567.
PMID
17082314
Source
pubmed
Published In
Blood
Volume
109
Issue
4
Publish Date
2007
Start Page
1559
End Page
1567
DOI
10.1182/blood-2006-05-020644

V(D)J recombinase-mediated processing of coding junctions at cryptic recombination signal sequences in peripheral T cells during human development.

V(D)J recombinase mediates rearrangements at immune loci and cryptic recombination signal sequences (cRSS), resulting in a variety of genomic rearrangements in normal lymphocytes and leukemic cells from children and adults. The frequency at which these rearrangements occur and their potential pathologic consequences are developmentally dependent. To gain insight into V(D)J recombinase-mediated events during human development, we investigated 265 coding junctions associated with cRSS sites at the hypoxanthine-guanine phosphoribosyltransferase (HPRT) locus in peripheral T cells from 111 children during the late stages of fetal development through early adolescence. We observed a number of specific V(D)J recombinase processing features that were both age and gender dependent. In particular, TdT-mediated nucleotide insertions varied depending on age and gender, including percentage of coding junctions containing N-nucleotide inserts, predominance of GC nucleotides, and presence of inverted repeats (Pr-nucleotides) at processed coding ends. In addition, the extent of exonucleolytic processing of coding ends was inversely related to age. We also observed a coding-partner-dependent difference in exonucleolytic processing and an age-specific difference in the subtypes of V(D)J-mediated events. We investigated these age- and gender-specific differences with recombination signal information content analysis of the cRSS sites in the human HPRT locus to gain insight into the mechanisms mediating these developmentally specific V(D)J recombinase-mediated rearrangements in humans.

Authors
Murray, JM; O'Neill, JP; Messier, T; Rivers, J; Walker, VE; McGonagle, B; Trombley, L; Cowell, LG; Kelsoe, G; McBlane, F; Finette, BA
MLA Citation
Murray, JM, O'Neill, JP, Messier, T, Rivers, J, Walker, VE, McGonagle, B, Trombley, L, Cowell, LG, Kelsoe, G, McBlane, F, and Finette, BA. "V(D)J recombinase-mediated processing of coding junctions at cryptic recombination signal sequences in peripheral T cells during human development." J Immunol 177.8 (October 15, 2006): 5393-5404.
PMID
17015725
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
177
Issue
8
Publish Date
2006
Start Page
5393
End Page
5404

Outside influence: TLRs direct hematopoietic cell fates.

Toll-like receptors (TLRs) modulate immune responses indirectly by promoting the efficacy of antigen presentation. In this issue of Immunity, Nagai et al. (2006) demonstrate that TLR signals also bias hematopoietic progenitor cells toward myelopoiesis directly by replacing cytokine and differentiative cues.

Authors
Holl, TM; Kelsoe, G
MLA Citation
Holl, TM, and Kelsoe, G. "Outside influence: TLRs direct hematopoietic cell fates." Immunity 24.6 (June 2006): 667-669. (Review)
PMID
16782021
Source
pubmed
Published In
Immunity
Volume
24
Issue
6
Publish Date
2006
Start Page
667
End Page
669
DOI
10.1016/j.immuni.2006.06.007

Inflammation and the reciprocal production of granulocytes and lymphocytes in bone marrow.

The coordinated production of leukocytes in bone marrow is crucial for innate and adaptive immunity. Inflammation alters normal leukocyte production by promoting granulopoiesis over lymphopoiesis, a response that supports the reactive neutrophilia that follows infection. Here we demonstrate that this specialization for granulopoiesis is determined by inflammation-induced reductions of growth and retention factors, most significantly stem cell factor and CXCL12, which act preferentially to inhibit lymphoid development. These hierarchical effects suggest that the normal equilibrium of leukocyte production in bone marrow is determined by lymphopoiesis' higher demand for specific growth factors and/or retention signals. Inflammation regulates this balance by reducing growth factors that have less impact on developing neutrophils than lymphocytes. We demonstrate that granulopoiesis and lymphopoiesis are coupled specifically in the bone marrow by development in a common niche and propose that the leukopoietic equilibrium is specified by limiting amounts of developmental resources.

Authors
Ueda, Y; Kondo, M; Kelsoe, G
MLA Citation
Ueda, Y, Kondo, M, and Kelsoe, G. "Inflammation and the reciprocal production of granulocytes and lymphocytes in bone marrow." J Exp Med 201.11 (June 6, 2005): 1771-1780.
Website
http://hdl.handle.net/10161/10906
PMID
15939792
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
201
Issue
11
Publish Date
2005
Start Page
1771
End Page
1780
DOI
10.1084/jem.20041419

Antibody polyspecificity and neutralization of HIV-1: a hypothesis.

HIV-1 has evolved many ways to evade protective host immune responses, thus creating a number of problems for HIV vaccine developers. In particular, durable, broadly specific neutralizing antibodies to HIV-1 have proved difficult to induce with current HIV-1 vaccine candidates. The recent observation that some broadly neutralizing anti-HIV-1 envelope monoclonal antibodies have polyspecific reactivities to host antigens have raised the hypothesis that one reason antibodies against some of the conserved HIV-1 envelope trimer neutralizing epitopes are not routinely made may be down-regulation of some specificities of anti-HIV-1 antibody producing B cells by host B cell tolerance mechanisms.

Authors
Haynes, BF; Moody, MA; Verkoczy, L; Kelsoe, G; Alam, SM
MLA Citation
Haynes, BF, Moody, MA, Verkoczy, L, Kelsoe, G, and Alam, SM. "Antibody polyspecificity and neutralization of HIV-1: a hypothesis." Hum Antibodies 14.3-4 (2005): 59-67. (Review)
PMID
16720975
Source
pubmed
Published In
Human antibodies
Volume
14
Issue
3-4
Publish Date
2005
Start Page
59
End Page
67

Computational tools for understanding sequence variability in recombination signals.

The recombination signals (RSs) that guide V(D)J rearrangement are remarkably diverse. In mice, fewer than 16% of RSs carry consensus heptamers and nonamers and none also contain a consensus spacer sequence. It is increasingly clear that this variability regulates recombination: genetic variability in RSs may help enforce allelic exclusion, determine the general nature of antigen receptor repertoires, and mitigate autoreactivity in B lymphocytes. The great diversity of RSs has largely precluded, however, empiric determinations of how RS sequence affects recombination. For example, 4(39) unique 23-RSs are possible or approximately 3 x 10(23) sequences; some 7 x 10(13) unique 23-RSs can be produced just by changes in the spacer. In contrast, the recombination activities of only 100 or so RSs have been measured, and it is unlikely that the activities of even a tiny fraction of extant RSs can be determined. We have addressed the problem of how sequence determines the efficiency of RS templates by generating computational models that describe the correlation structure of mouse RSs. These models successfully predict RS activity and identify functional, cryptic RSs (cRSs). These models permit studies to identify RSs and cRSs for empiric study and constitute a tool useful for understanding RS structure and function.

Authors
Cowell, LG; Davila, M; Ramsden, D; Kelsoe, G
MLA Citation
Cowell, LG, Davila, M, Ramsden, D, and Kelsoe, G. "Computational tools for understanding sequence variability in recombination signals." Immunol Rev 200 (August 2004): 57-69. (Review)
PMID
15242396
Source
pubmed
Published In
Immunological Reviews
Volume
200
Publish Date
2004
Start Page
57
End Page
69
DOI
10.1111/j.0105-2896.2004.00171.x

Should we B-leavin' now?

Authors
Gunn, MD; Kelsoe, G
MLA Citation
Gunn, MD, and Kelsoe, G. "Should we B-leavin' now?." Nat Immunol 5.7 (July 2004): 703-704.
PMID
15224099
Source
pubmed
Published In
Nature Immunology
Volume
5
Issue
7
Publish Date
2004
Start Page
703
End Page
704
DOI
10.1038/ni0704-703

Inflammation controls B lymphopoiesis by regulating chemokine CXCL12 expression.

Inflammation removes developing and mature lymphocytes from the bone marrow (BM) and induces the appearance of developing B cells in the spleen. BM granulocyte numbers increase after lymphocyte reductions to support a reactive granulocytosis. Here, we demonstrate that inflammation, acting primarily through tumor necrosis factor alpha (TNFalpha), mobilizes BM lymphocytes. Mobilization reflects a reduced CXCL12 message and protein in BM and changes to the BM environment that prevents homing by cells from naive donors. The effects of TNFalpha are potentiated by interleukin 1 beta (IL-1beta), which acts primarily to expand the BM granulocyte compartment. Our observations indicate that inflammation induces lymphocyte mobilization by suppressing CXCL12 retention signals in BM, which, in turn, increases the ability of IL-1beta to expand the BM granulocyte compartment. Consistent with this idea, lymphocyte mobilization and a modest expansion of BM granulocyte numbers follow injections of pertussis toxin. We propose that TNFalpha and IL-1beta transiently specialize the BM to support acute granulocytic responses and consequently promote extramedullary lymphopoiesis.

Authors
Ueda, Y; Yang, K; Foster, SJ; Kondo, M; Kelsoe, G
MLA Citation
Ueda, Y, Yang, K, Foster, SJ, Kondo, M, and Kelsoe, G. "Inflammation controls B lymphopoiesis by regulating chemokine CXCL12 expression." J Exp Med 199.1 (January 5, 2004): 47-58.
Website
http://hdl.handle.net/10161/10909
PMID
14707114
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
199
Issue
1
Publish Date
2004
Start Page
47
End Page
58
DOI
10.1084/jem.20031104

A birthday for B cells: Lymphopoiesis II, a scientific symposium honoring Max Cooper.

Authors
Kincade, PW; Kelsoe, G
MLA Citation
Kincade, PW, and Kelsoe, G. "A birthday for B cells: Lymphopoiesis II, a scientific symposium honoring Max Cooper." Nat Immunol 4.12 (December 2003): 1155-1157.
PMID
14639461
Source
pubmed
Published In
Nature Immunology
Volume
4
Issue
12
Publish Date
2003
Start Page
1155
End Page
1157
DOI
10.1038/ni1203-1155

Transcriptome analysis of mouse stem cells and early embryos.

Understanding and harnessing cellular potency are fundamental in biology and are also critical to the future therapeutic use of stem cells. Transcriptome analysis of these pluripotent cells is a first step towards such goals. Starting with sources that include oocytes, blastocysts, and embryonic and adult stem cells, we obtained 249,200 high-quality EST sequences and clustered them with public sequences to produce an index of approximately 30,000 total mouse genes that includes 977 previously unidentified genes. Analysis of gene expression levels by EST frequency identifies genes that characterize preimplantation embryos, embryonic stem cells, and adult stem cells, thus providing potential markers as well as clues to the functional features of these cells. Principal component analysis identified a set of 88 genes whose average expression levels decrease from oocytes to blastocysts, stem cells, postimplantation embryos, and finally to newborn tissues. This can be a first step towards a possible definition of a molecular scale of cellular potency. The sequences and cDNA clones recovered in this work provide a comprehensive resource for genes functioning in early mouse embryos and stem cells. The nonrestricted community access to the resource can accelerate a wide range of research, particularly in reproductive and regenerative medicine.

Authors
Sharov, AA; Piao, Y; Matoba, R; Dudekula, DB; Qian, Y; VanBuren, V; Falco, G; Martin, PR; Stagg, CA; Bassey, UC; Wang, Y; Carter, MG; Hamatani, T; Aiba, K; Akutsu, H; Sharova, L; Tanaka, TS; Kimber, WL; Yoshikawa, T; Jaradat, SA; Pantano, S; Nagaraja, R; Boheler, KR; Taub, D; Hodes, RJ; Longo, DL; Schlessinger, D; Keller, J; Klotz, E; Kelsoe, G; Umezawa, A; Vescovi, AL; Rossant, J; Kunath, T; Hogan, BLM; Curci, A; D'Urso, M; Kelso, J; Hide, W; Ko, MSH
MLA Citation
Sharov, AA, Piao, Y, Matoba, R, Dudekula, DB, Qian, Y, VanBuren, V, Falco, G, Martin, PR, Stagg, CA, Bassey, UC, Wang, Y, Carter, MG, Hamatani, T, Aiba, K, Akutsu, H, Sharova, L, Tanaka, TS, Kimber, WL, Yoshikawa, T, Jaradat, SA, Pantano, S, Nagaraja, R, Boheler, KR, Taub, D, Hodes, RJ, Longo, DL, Schlessinger, D, Keller, J, Klotz, E, Kelsoe, G, Umezawa, A, Vescovi, AL, Rossant, J, Kunath, T, Hogan, BLM, Curci, A, D'Urso, M, Kelso, J, Hide, W, and Ko, MSH. "Transcriptome analysis of mouse stem cells and early embryos." PLoS Biol 1.3 (December 2003): E74-.
PMID
14691545
Source
pubmed
Published In
PLoS biology
Volume
1
Issue
3
Publish Date
2003
Start Page
E74
DOI
10.1371/journal.pbio.0000074

Therapeutic CD154 antibody for lupus: promise for the future?

Systemic lupus erythematosus (SLE) is a prototypical systemic autoimmune disease characterized by the production of pathogenic autoantibodies. A new study demonstrates that passive antibody specific for the TNF family member, CD154, ameliorates disease by reducing levels of self-reactive antibody in the serum. This study demonstrates a substantial potential for anti-CD154 antibody in the treatment of humoral autoimmunity.

Authors
Kelsoe, G
MLA Citation
Kelsoe, G. "Therapeutic CD154 antibody for lupus: promise for the future?." J Clin Invest 112.10 (November 2003): 1480-1482.
PMID
14617748
Source
pubmed
Published In
Journal of Clinical Investigation
Volume
112
Issue
10
Publish Date
2003
Start Page
1480
End Page
1482
DOI
10.1172/JCI20371

A functional analysis of the spacer of V(D)J recombination signal sequences.

During lymphocyte development, V(D)J recombination assembles antigen receptor genes from component V, D, and J gene segments. These gene segments are flanked by a recombination signal sequence (RSS), which serves as the binding site for the recombination machinery. The murine Jbeta2.6 gene segment is a recombinationally inactive pseudogene, but examination of its RSS reveals no obvious reason for its failure to recombine. Mutagenesis of the Jbeta2.6 RSS demonstrates that the sequences of the heptamer, nonamer, and spacer are all important. Strikingly, changes solely in the spacer sequence can result in dramatic differences in the level of recombination. The subsequent analysis of a library of more than 4,000 spacer variants revealed that spacer residues of particular functional importance are correlated with their degree of conservation. Biochemical assays indicate distinct cooperation between the spacer and heptamer/nonamer along each step of the reaction pathway. The results suggest that the spacer serves not only to ensure the appropriate distance between the heptamer and nonamer but also regulates RSS activity by providing additional RAG:RSS interaction surfaces. We conclude that while RSSs are defined by a "digital" requirement for absolutely conserved nucleotides, the quality of RSS function is determined in an "analog" manner by numerous complex interactions between the RAG proteins and the less-well conserved nucleotides in the heptamer, the nonamer, and, importantly, the spacer. Those modulatory effects are accurately predicted by a new computational algorithm for "RSS information content." The interplay between such binary and multiplicative modes of interactions provides a general model for analyzing protein-DNA interactions in various biological systems.

Authors
Lee, AI; Fugmann, SD; Cowell, LG; Ptaszek, LM; Kelsoe, G; Schatz, DG
MLA Citation
Lee, AI, Fugmann, SD, Cowell, LG, Ptaszek, LM, Kelsoe, G, and Schatz, DG. "A functional analysis of the spacer of V(D)J recombination signal sequences." PLoS Biol 1.1 (October 2003): E1-.
Website
http://hdl.handle.net/10161/11485
PMID
14551903
Source
pubmed
Published In
PLoS biology
Volume
1
Issue
1
Publish Date
2003
Start Page
E1
DOI
10.1371/journal.pbio.0000001

Enhanced differentiation of splenic plasma cells but diminished long-lived high-affinity bone marrow plasma cells in aged mice.

In the present work, we have dissected the mechanisms responsible for the impaired humoral responses in aging. We found that there was a substantially higher level of Ab-forming cells in the spleens of aged mice than that of young controls. However, the number of high-affinity, class-switched Ab-forming cells was severely decreased in the spleen of aged mice. The accumulation of low-affinity IgM Ab-forming cells in the spleens of aged animals was not due to a deficiency in isotype switching because the number of total IgG1 splenic plasma cells was not significantly reduced. Remarkably, plasma cells of both low and high affinity were significantly diminished in the bone marrow of aged mice compared with that of young mice. The results from reconstitution experiments showed that aged bone marrow was less supportive for plasma cells derived from young splenic B cells. These findings suggest that humoral immune deficiency in aging results from at least two mechanisms: the inability to generate sufficient numbers of high-affinity Ab-forming cells, which is a result of diminished germinal center reaction, and the defective bone marrow environment that has diminished ability to support the selection and survival of long-term Ab-forming cells.

Authors
Han, S; Yang, K; Ozen, Z; Peng, W; Marinova, E; Kelsoe, G; Zheng, B
MLA Citation
Han, S, Yang, K, Ozen, Z, Peng, W, Marinova, E, Kelsoe, G, and Zheng, B. "Enhanced differentiation of splenic plasma cells but diminished long-lived high-affinity bone marrow plasma cells in aged mice." J Immunol 170.3 (February 1, 2003): 1267-1273.
PMID
12538685
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
170
Issue
3
Publish Date
2003
Start Page
1267
End Page
1273

Prospective estimation of recombination signal efficiency and identification of functional cryptic signals in the genome by statistical modeling.

The recombination signals (RS) that guide V(D)J recombination are phylogenetically conserved but retain a surprising degree of sequence variability, especially in the nonamer and spacer. To characterize RS variability, we computed the position-wise information, a measure correlated with sequence conservation, for each nucleotide position in an RS alignment and demonstrate that most position-wise information is present in the RS heptamers and nonamers. We have previously demonstrated significant correlations between RS positions and here show that statistical models of the correlation structure that underlies RS variability efficiently identify physiologic and cryptic RS and accurately predict the recombination efficiencies of natural and synthetic RS. In scans of mouse and human genomes, these models identify a highly conserved family of repetitive DNA as an unexpected source of frequent, cryptic RS that rearrange both in extrachromosomal substrates and in their genomic context.

Authors
Cowell, LG; Davila, M; Yang, K; Kepler, TB; Kelsoe, G
MLA Citation
Cowell, LG, Davila, M, Yang, K, Kepler, TB, and Kelsoe, G. "Prospective estimation of recombination signal efficiency and identification of functional cryptic signals in the genome by statistical modeling." J Exp Med 197.2 (January 20, 2003): 207-220.
Website
http://hdl.handle.net/10161/10907
PMID
12538660
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
197
Issue
2
Publish Date
2003
Start Page
207
End Page
220

The "dispensable" portion of RAG2 is necessary for efficient V-to-DJ rearrangement during B and T cell development.

Previous in vitro studies defined the minimal regions of RAG1 and RAG2 essential for V(D)J recombination. In order to characterize the role of the C-terminal "dispensable" portion of RAG2, we generated core-RAG2 knock-in mice. We found that the core-RAG2-containing recombinase complex is selectively defective in catalyzing V-to-DJ rearrangement at the IgH and TCRbeta loci, resulting in partial developmental blocks in B and T lymphopoiesis. Analysis of recombination intermediates showed defects at the cleavage phase of the reaction. We also observed a reduction in overall recombinase activity in core-RAG2-expressing thymocytes, leading us to suggest that the interaction of a defective recombinase with RSS sequences unique to VH and Vbeta gene segments may underlie the specific V-to-DJ rearrangement defect in core-RAG2 mice.

Authors
Liang, H-E; Hsu, L-Y; Cado, D; Cowell, LG; Kelsoe, G; Schlissel, MS
MLA Citation
Liang, H-E, Hsu, L-Y, Cado, D, Cowell, LG, Kelsoe, G, and Schlissel, MS. "The "dispensable" portion of RAG2 is necessary for efficient V-to-DJ rearrangement during B and T cell development." Immunity 17.5 (November 2002): 639-651.
PMID
12433370
Source
pubmed
Published In
Immunity
Volume
17
Issue
5
Publish Date
2002
Start Page
639
End Page
651

Very low affinity B cells form germinal centers, become memory B cells, and participate in secondary immune responses when higher affinity competition is reduced.

To understand the relationship between the affinity of the B cell antigen receptor (BCR) and the immune response to antigen, two lines of immunoglobulin H chain transgenic (Tg) mice were created. H50Gmu(a) and T1(V23)mu(a) mice express mu H chain transgenes that associate with the lambda1 L chains to bind the (4-hydroxy-3-nitrophenyl)acetyl hapten with association constants (K(a)s) of only 1.2 x 10(5) M(-1) and 3 x 10(4) M(-1), respectively. Both lines mounted substantial antibody-forming cell (AFC) and germinal center (GC) responses. H50Gmu(a) Tg mice also generated memory B cells. T1(V23)mu(a) B cells formed AFC and GCs, but were largely replaced in late GCs by antigen-specific cells that express endogenous BCRs. Thus, B lymphocytes carrying BCRs with affinities previously thought to be irrelevant in specific immune responses are in fact capable of complete T cell-dependent immune responses when relieved of substantial competition from other B cells. The failure to observe such B cells normally in late primary responses and in memory B cell populations is the result of competition, rather than an intrinsic inability of low affinity B cells.

Authors
Dal Porto, JM; Haberman, AM; Kelsoe, G; Shlomchik, MJ
MLA Citation
Dal Porto, JM, Haberman, AM, Kelsoe, G, and Shlomchik, MJ. "Very low affinity B cells form germinal centers, become memory B cells, and participate in secondary immune responses when higher affinity competition is reduced." J Exp Med 195.9 (May 6, 2002): 1215-1221.
Website
http://hdl.handle.net/10161/10908
PMID
11994427
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
195
Issue
9
Publish Date
2002
Start Page
1215
End Page
1221

Identification and utilization of arbitrary correlations in models of recombination signal sequences.

BACKGROUND: A significant challenge in bioinformatics is to develop methods for detecting and modeling patterns in variable DNA sequence sites, such as protein-binding sites in regulatory DNA. Current approaches sometimes perform poorly when positions in the site do not independently affect protein binding. We developed a statistical technique for modeling the correlation structure in variable DNA sequence sites. The method places no restrictions on the number of correlated positions or on their spatial relationship within the site. No prior empirical evidence for the correlation structure is necessary. RESULTS: We applied our method to the recombination signal sequences (RSS) that direct assembly of B-cell and T-cell antigen-receptor genes via V(D)J recombination. The technique is based on model selection by cross-validation and produces models that allow computation of an information score for any signal-length sequence. We also modeled RSS using order zero and order one Markov chains. The scores from all models are highly correlated with measured recombination efficiencies, but the models arising from our technique are better than the Markov models at discriminating RSS from non-RSS. CONCLUSIONS: Our model-development procedure produces models that estimate well the recombinogenic potential of RSS and are better at RSS recognition than the order zero and order one Markov models. Our models are, therefore, valuable for studying the regulation of both physiologic and aberrant V(D)J recombination. The approach could be equally powerful for the study of promoter and enhancer elements, splice sites, and other DNA regulatory sites that are highly variable at the level of individual nucleotide positions.

Authors
Cowell, LG; Davila, M; Kepler, TB; Kelsoe, G
MLA Citation
Cowell, LG, Davila, M, Kepler, TB, and Kelsoe, G. "Identification and utilization of arbitrary correlations in models of recombination signal sequences." Genome Biol 3.12 (2002): RESEARCH0072-.
Website
http://hdl.handle.net/10161/11484
PMID
12537561
Source
pubmed
Published In
Genome Biology
Volume
3
Issue
12
Publish Date
2002
Start Page
RESEARCH0072

Spontaneous formation of germinal centers in autoimmune mice.

The mechanisms of autoantibody production are not well understood. Germinal centers (GC) may be important sites of immune disregulation in autoimmune diseases. In this study, we document the presence of spontaneous GC formation in the spleens of several autoimmune mouse strains that spontaneously develop autoimmune Type I diabetes and a lupus-like disease. In contrast, mouse strains that do not develop lupus did not exhibit spontaneous formation of GC. In all of the autoimmune strains studied, GC were present at 1-2 months of age, a time that closely parallels the appearance of autoantibodies. Like the GC that develop after purposeful immunization, GC in autoimmune mice contained B220(+), PNA(+), and GL-7(+) B cells, and FDC-M1(+) follicular dendritic cells. In addition, spontaneously formed GC in autoimmunity and those caused by immunization were abrogated in a similar way by a short-term treatment with anti-CD40 ligand antibody. These data indicate that spontaneously forming GC in autoimmunity are similar to those appearing after purposeful immunization.

Authors
Luzina, IG; Atamas, SP; Storrer, CE; daSilva, LC; Kelsoe, G; Papadimitriou, JC; Handwerger, BS
MLA Citation
Luzina, IG, Atamas, SP, Storrer, CE, daSilva, LC, Kelsoe, G, Papadimitriou, JC, and Handwerger, BS. "Spontaneous formation of germinal centers in autoimmune mice." J Leukoc Biol 70.4 (October 2001): 578-584.
PMID
11590194
Source
pubmed
Published In
Journal of leukocyte biology
Volume
70
Issue
4
Publish Date
2001
Start Page
578
End Page
584

Definition of a novel cellular constituent of the bone marrow that regulates the response of immature B cells to B cell antigen receptor engagement.

Previously we defined a Thy1(dull) bone marrow-derived cell population that regulated fate decisions by immature B cells after Ag receptor signaling. The microenvironmental signals provided by this cell population were shown to redirect the B cell Ag receptor -induced apoptotic response of immature B cells toward continued recombination-activating gene (RAG) expression and secondary light chain recombination (receptor editing). Neither the identity of the cell responsible for this activity nor its role in immature B cell development in vivo were addressed by these previous studies. Here we show that this protective microenvironmental niche is defined by the presence of a novel Thy1(dull), DX5(pos) cell that can be found in close association with immature B cells in vivo. Depletion of this cell eliminates the anti-apoptotic effect of bone marrow in vitro and leads to a significant decrease in the number and frequency of bone marrow immature B cells in vivo. We propose that, just as the bone marrow environment is essential for the survival and progression of pro-B and pre-B cells through their respective developmental checkpoints, this cellular niche regulates the progression of immature stage B cells through negative selection.

Authors
Sandel, PC; Gendelman, M; Kelsoe, G; Monroe, JG
MLA Citation
Sandel, PC, Gendelman, M, Kelsoe, G, and Monroe, JG. "Definition of a novel cellular constituent of the bone marrow that regulates the response of immature B cells to B cell antigen receptor engagement." J Immunol 166.10 (May 15, 2001): 5935-5944.
PMID
11342608
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
166
Issue
10
Publish Date
2001
Start Page
5935
End Page
5944

CD4+Thy1- thymocytes with a Th-type 2 cytokine response.

We have identified a small subset of CD3(+), CD4(+), CD8(-) thymocytes that do not express Thy1 (CD90). This Thy1(-) subset represents 1-3.7% of the total number of thymocytes in a naive mouse. CD4(+)Thy1(-) thymocytes express high levels of CD3, intermediate to high levels of heat-stable antigen (HSA), and low levels of CD25, CD45RB, CD69, CD44 and CD62L. They produce high titers of IL-4 and no IFN-gamma upon stimulation in vitro, a response characteristic of T(h)2 cells. In the thymi of mice infected neonatally with a high dose of the retrovirus Cas-Br-E MuLV, the frequency of CD4(+)Thy1(-) cells increased approximately 10-fold. High-dose virus infection resulted in decreased HSA and increased CD44 expression on CD4(+)Thy1(-) cells relative to cells from naive mice. CD4(+)Thy1(-) cells from high-dose infected mice also secreted IL-4 and not IFN-gamma upon in vitro stimulation. We previously reported that infection of newborn mice with a high dose of murine retrovirus results in the induction of a non-protective anti-viral T(h)2 T cell response; CD4(+)Thy1(-) thymocytes with a T(h)2-like cytokine profile may play a role in determining the cytokine bias of this anti-viral response.

Authors
Cerasoli, DM; Kelsoe, G; Sarzotti, M
MLA Citation
Cerasoli, DM, Kelsoe, G, and Sarzotti, M. "CD4+Thy1- thymocytes with a Th-type 2 cytokine response." Int Immunol 13.1 (January 2001): 75-83.
PMID
11133836
Source
pubmed
Published In
International Immunology
Volume
13
Issue
1
Publish Date
2001
Start Page
75
End Page
83

A role for secondary V(D)J recombination in oncogenic chromosomal translocations?

Chromosomal translocations are hallmarks of certain lymphoproliferative disorders. Indeed, in many leukemias and lymphomas, translocations are the transforming event that brings about malignancy. Recurrence of the immunoglobulin (Ig) and T-cell receptor (Tcr) loci at the breakpoints of oncogenic chromosomal translocations has led to speculation that the lymphocyte-specific process of V(D)J rearrangement, which is necessary for the generation of functional Ig and TCR antigen receptors on B and T lymphocytes, mediates translocation. Recent studies have led to a fuller understanding of the molecular mechanisms of V(D)J rearrangement and have revealed that the V(D)J recombinase possesses latent transposase activity. These studies have led to plausible models of illegitimate V(D)J recombination producing chromosomal translocations consistent with those present in lymphomas and leukemias. Errors of V(D)J recombination may even generate lymphomas with the phenotypes of mature cells. For example, follicular and Burkitt's lymphomas have been classified by phenotype and somatic genotype as malignant germinal center (GC) B or post-GC B cells. The GC is a site of affinity maturation where B cells undergo V(D)J hypermutation and Ig class switch; in addition, much evidence has accumulated to suggest that GC B cells may also support secondary V(D)J recombination. Interestingly, all three of these elements, genomic plasticity, mutation, and translocation breakpoints near switch sites or recombinational elements, are characteristic of certain lymphomas. The high frequency of lymphomas carrying these GC markers suggests that the GC reaction may play a significant role in lymphomagenesis.

Authors
Davila, M; Foster, S; Kelsoe, G; Yang, K
MLA Citation
Davila, M, Foster, S, Kelsoe, G, and Yang, K. "A role for secondary V(D)J recombination in oncogenic chromosomal translocations?." Adv Cancer Res 81 (2001): 61-92. (Review)
PMID
11430596
Source
pubmed
Published In
Advances in cancer research
Volume
81
Publish Date
2001
Start Page
61
End Page
92

CD4+ Thy1- thymocytes with a Th-type 2 cytokine response

We have identified a small subset of CD3+, CD4+, CD8- thymocytes that do not express Thy1 (CD90). This Thy1- subset represents 1-3.7% of the total number of thymocytes in a naive mouse. CD4+ Thy1- thymocytes express high levels of CD3, intermediate to high levels of heat-stable antigen (HSA), and low levels of CD25, CD45RB, CD69, CD44 and CD62L. They produce high titers of IL-4 and no IFN-γ upon stimulation in vitro, a response characteristic of Th2 cells. In the thymi of mice infected neonatally with a high dose of the retrovirus Cas-Br-E MuLV, the frequency of CD4+Thy1- cells increased ∼10-fold. High-dose virus infection resulted in decreased HSA and increased CD44 expression on CD4+Thy1- cells relative to cells from naive mice. CD4+Thy1- cells from high-dose infected mice also secreted IL-4 and not IFN-γ upon in vitro stimulation. We previously reported that infection of newborn mice with a high dose of murine retrovirus results in the induction of a non-protective anti-viral Th2 T cell response; CD4+Thy1- thymocytes with a Th2-like cytokine profile may play a role in determining the cytokine bias of this anti-viral response.

Authors
Cerasoli, DM; Kelsoe, G; Sarzotti, M
MLA Citation
Cerasoli, DM, Kelsoe, G, and Sarzotti, M. "CD4+ Thy1- thymocytes with a Th-type 2 cytokine response." International Immunology 13.1 (2001): 75-83.
Source
scival
Published In
International Immunology
Volume
13
Issue
1
Publish Date
2001
Start Page
75
End Page
83

Complement C4 inhibits systemic autoimmunity through a mechanism independent of complement receptors CR1 and CR2.

The complement system enhances antibody responses to T-dependent antigens, but paradoxically, deficiencies in C1 and C4 are strongly linked to autoantibody production in humans. In mice, disruption of the C1qa gene also results in spontaneous autoimmunity. Moreover, deficiencies in C4 or complement receptors 1 and 2 (CR1/CR2) lead to reduced selection against autoreactive B cells and impaired humoral responses. These observations suggest that C1 and C4 act through CR1/CR2 to enhance humoral immunity and somehow suppress autoimmunity. Here we report high titers of spontaneous antinuclear antibody (ANA) in C4(-/)- mice. This systemic lupus erythematosus-like autoimmunity is highly penetrant; by 10 mo of age, all C4(-)(/)- females and most males produced ANA. In contrast, titers and frequencies of ANA in Cr2(-)(/)- mice, which are deficient in CR1 and CR2, never rose significantly above those in normal controls. Glomerular deposition of immune complexes (ICs), glomerulonephritis, and splenomegaly were observed in C4(-)(/)- but not Cr2(-)(/)- mice. C4(-)(/)-, but not Cr2(-)(/)-, mice accumulate activated T and B cells. Clearance of circulating ICs is impaired in preautoimmune C4(-)(/)-, but not Cr2(-)(/)-, mice. C4 deficiency causes spontaneous, lupus-like autoimmunity through a mechanism that is independent of CR1/CR2.

Authors
Chen, Z; Koralov, SB; Kelsoe, G
MLA Citation
Chen, Z, Koralov, SB, and Kelsoe, G. "Complement C4 inhibits systemic autoimmunity through a mechanism independent of complement receptors CR1 and CR2." J Exp Med 192.9 (November 6, 2000): 1339-1352.
Website
http://hdl.handle.net/10161/11489
PMID
11067882
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
192
Issue
9
Publish Date
2000
Start Page
1339
End Page
1352

Remembrance of things past.

Authors
Kelsoe, G
MLA Citation
Kelsoe, G. "Remembrance of things past." Nat Immunol 1.5 (November 2000): 375-376.
PMID
11062494
Source
pubmed
Published In
Nature Immunology
Volume
1
Issue
5
Publish Date
2000
Start Page
375
End Page
376
DOI
10.1038/80811

Regulation of humoral immune responses by CD21/CD35.

Before antigen-specific immunity arises, the complement system responds by activation through the classical and/or alternative pathways leading to the covalent deposition of complement fragments. Three models, not mutually exclusive, have been proposed to explain how these complement fragments interact with their receptors, CD21/CD35, to enhance humoral immune responses: i) CD21/CD35 retain and focus antigens for optimal presentation; ii) CD21/CD35 on B cells serve as enhancing co-receptors for B-cell antigen receptor (BCR) signaling; iii) CD21/CD35 regulate B-cell responses, by CD19 aggregation. The coreceptor model led us to predict that CD21/CD3 5 may lower the threshold of BCR affinity to diversify the repertoire of humoral immune responses, but surprisingly, CD21/CD3 5-deficient mice expressing a transgenic BCR with very low affinity (Kalpha approximately =1.2 x 10(5) M(-1)) for the (4-hydroxy-3-nitrophenyl)acetyl hapten generated significant antibody and germinal center responses to even low doses of antigens in alum. The magnitudes of these responses were much below those of normal controls but higher doses of antigens substantially rectified these deficits. Thus, while CD21/CD35 play important roles in humoral immunity, our observations provide little support to the hypothesis that CD21/CD35 promote clonal diversity in immune responses by helping recruit low-affinity B cells.

Authors
Chen, Z; Koralov, SB; Kelsoe, G
MLA Citation
Chen, Z, Koralov, SB, and Kelsoe, G. "Regulation of humoral immune responses by CD21/CD35." Immunol Rev 176 (August 2000): 194-204. (Review)
PMID
11043778
Source
pubmed
Published In
Immunological Reviews
Volume
176
Publish Date
2000
Start Page
194
End Page
204

Humoral immune responses in Cr2-/- mice: enhanced affinity maturation but impaired antibody persistence.

Deficiency in CD21/CD35 by disruption of the Cr2 loci leads to impaired humoral immune responses. In this study, we detail the role of CD21/CD35 on Ab responses to the hapten (4-hydroxy-3-nitrophenyl)acetyl conjugated to chicken gamma-globulin. Surprisingly, Cr2-/- mice generate significant Ab responses and germinal center (GC) reactions to low doses of this Ag in alum, although the magnitude of their responses is much reduced in comparison with those of Cr2+/- and C57BL/6 controls. Increasing Ag dose partially corrected this deficit. In situ study of the somatic genetics of GC B cells demonstrated that VDJ hypermutation does not require CD21/CD35, and Cr2-/- mice exhibited enhanced affinity maturation of serum Ab in the post-GC phase of the primary response. On the other hand, Cr2-/- mice displayed accelerated loss of serum Ab and long-lived Ab-forming cells. These observations suggest that B cell activation/survival signals mediated by CD21 and/or the retention of Ag by CD21/CD35 play important roles in the generation, quality, and maintenance of serum Ab.

Authors
Chen, Z; Koralov, SB; Gendelman, M; Carroll, MC; Kelsoe, G
MLA Citation
Chen, Z, Koralov, SB, Gendelman, M, Carroll, MC, and Kelsoe, G. "Humoral immune responses in Cr2-/- mice: enhanced affinity maturation but impaired antibody persistence." J Immunol 164.9 (May 1, 2000): 4522-4532.
PMID
10779753
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
164
Issue
9
Publish Date
2000
Start Page
4522
End Page
4532

Germline sequences of V(H)7183 gene family members in C57BL/6 mice demonstrate natural selection of particular sequences during recent evolution.

Authors
Langdon, SD; Inaioki, M; Kelsoe, G; Tedder, TF
MLA Citation
Langdon, SD, Inaioki, M, Kelsoe, G, and Tedder, TF. "Germline sequences of V(H)7183 gene family members in C57BL/6 mice demonstrate natural selection of particular sequences during recent evolution." Immunogenetics 51.3 (March 2000): 241-245.
PMID
10752635
Source
pubmed
Published In
Immunogenetics
Volume
51
Issue
3
Publish Date
2000
Start Page
241
End Page
245

Studies of the humoral immune response.

The humoral immune response arises from a complex choreography of cells and molecules that interact to produce lasting and effective defenses against pathogens. For more than fifteen years, our laboratory has studied how humoral responses are initiated, how they mature, and how they are remembered. This work has come from many hands and in this brief synopsis, I cannot provide the full recognition that my students, postdoctoral fellows, and collaborators merit. I hope that my colleagues can accept this translucence and know that their efforts are recognized and deeply appreciated, nonetheless.

Authors
Kelsoe, G
MLA Citation
Kelsoe, G. "Studies of the humoral immune response." Immunol Res 22.2-3 (2000): 199-210. (Review)
PMID
11339356
Source
pubmed
Published In
Immunologic Research
Volume
22
Issue
2-3
Publish Date
2000
Start Page
199
End Page
210
DOI
10.1385/IR:22:2-3:199

Relaxed negative selection in germinal centers and impaired affinity maturation in bcl-xL transgenic mice.

The role of apoptosis in affinity maturation was investigated by determining the affinity of (4-hydroxy-3-nitrophenyl)acetyl (NP)-specific antibody-forming cells (AFCs) and serum antibody in transgenic mice that overexpress a suppressor of apoptosis, Bcl-xL, in the B cell compartment. Although transgenic animals briefly expressed higher numbers of splenic AFCs after immunization, the bcl-xL transgene did not increase the number or size of germinal centers (GCs), alter the levels of serum antibody, or change the frequency of NP-specific, long-lived AFCs. Nonetheless, the bcl-xL transgene product, in addition to endogenous Bcl-xL, reduced apoptosis in GC B cells and resulted in the expansion of B lymphocytes bearing VDJ rearrangements that are usually rare in primary anti-NP responses. Long-lived AFCs bearing these noncanonical rearrangements were frequent in the bone marrow and secreted immunoglobulin G(1) antibodies with low affinity for NP. The abundance of noncanonical cells lowered the average affinity of long-lived AFCs and serum antibody, demonstrating that Bcl-xL and apoptosis influence clonal selection/maintenance for affinity maturation.

Authors
Takahashi, Y; Cerasoli, DM; Dal Porto, JM; Shimoda, M; Freund, R; Fang, W; Telander, DG; Malvey, EN; Mueller, DL; Behrens, TW; Kelsoe, G
MLA Citation
Takahashi, Y, Cerasoli, DM, Dal Porto, JM, Shimoda, M, Freund, R, Fang, W, Telander, DG, Malvey, EN, Mueller, DL, Behrens, TW, and Kelsoe, G. "Relaxed negative selection in germinal centers and impaired affinity maturation in bcl-xL transgenic mice." J Exp Med 190.3 (August 2, 1999): 399-410.
Website
http://hdl.handle.net/10161/10897
PMID
10430628
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
190
Issue
3
Publish Date
1999
Start Page
399
End Page
410

RAG2:GFP knockin mice reveal novel aspects of RAG2 expression in primary and peripheral lymphoid tissues.

We generated mice in which a functional RAG2:GFP fusion gene is knocked in to the endogenous RAG2 locus. In bone marrow and thymus, RAG2:GFP expression occurs in appropriate stages of developing B and T cells as well as in immature bone marrow IgM+ B cells. RAG2:GFP also is expressed in IgD+ B cells following cross-linking of IgM on immature IgM+ IgD+ B cells generated in vitro. RAG2:GFP expression is undetectable in most immature splenic B cells; however, in young RAG2:GFP mice, there are substantial numbers of splenic RAG2:GFP+ cells that mostly resemble pre-B cells. The latter population decreases in size with age but reappears following immunization of older RAG2:GFP mice. We discuss the implications of these findings for current models of receptor assembly and diversification.

Authors
Monroe, RJ; Seidl, KJ; Gaertner, F; Han, S; Chen, F; Sekiguchi, J; Wang, J; Ferrini, R; Davidson, L; Kelsoe, G; Alt, FW
MLA Citation
Monroe, RJ, Seidl, KJ, Gaertner, F, Han, S, Chen, F, Sekiguchi, J, Wang, J, Ferrini, R, Davidson, L, Kelsoe, G, and Alt, FW. "RAG2:GFP knockin mice reveal novel aspects of RAG2 expression in primary and peripheral lymphoid tissues." Immunity 11.2 (August 1999): 201-212.
PMID
10485655
Source
pubmed
Published In
Immunity
Volume
11
Issue
2
Publish Date
1999
Start Page
201
End Page
212

Spontaneous germinal center formation in autoimmune mice.

Authors
Luzina, IG; daSilva, LC; Kelsoe, GH; Gerdelman, M; Storrer, CE; Handwerger, BS
MLA Citation
Luzina, IG, daSilva, LC, Kelsoe, GH, Gerdelman, M, Storrer, CE, and Handwerger, BS. "Spontaneous germinal center formation in autoimmune mice." FASEB JOURNAL 13.5 (March 15, 1999): A956-A956.
Source
wos-lite
Published In
The FASEB journal : official publication of the Federation of American Societies for Experimental Biology
Volume
13
Issue
5
Publish Date
1999
Start Page
A956
End Page
A956

V(D)J hypermutation and receptor revision: coloring outside the lines.

At least three mechanisms increase potential genetic diversity in peripheral B lymphocytes: hypermutation, gene conversion and secondary V(D)J rearrangements. These diversifying activities were once believed to be strictly confined to the immunoglobulin loci and B cells. Recent experiments demonstrate that this is not the case. Hypermutation has now been shown to diversify the BCL-6 genes of germinal-center B cells. The role, if any, of these mutations in the immune response remains unknown but the notion that the hypermutation mechanism is targeted solely to immunoglobulin genes is no longer tenable. Peripheral T cells may also diversify their antigen receptors by the reactivation of RAG (recombination-activating gene)1 and RAG2 and secondary V(D)J rearrangements. These new findings suggest a remarkable genetic plasticity in subsets of antigen-reactive lymphocytes and may frame new questions of clonal selection and self tolerance.

Authors
Kelsoe, G
MLA Citation
Kelsoe, G. "V(D)J hypermutation and receptor revision: coloring outside the lines." Curr Opin Immunol 11.1 (February 1999): 70-75. (Review)
PMID
10047542
Source
pubmed
Published In
Current Opinion in Immunology
Volume
11
Issue
1
Publish Date
1999
Start Page
70
End Page
75

Do germinal centers have a role in the generation of lymphomas?

Authors
Yang, K; Davila, M; Kelsoe, G
MLA Citation
Yang, K, Davila, M, and Kelsoe, G. "Do germinal centers have a role in the generation of lymphomas?." Curr Top Microbiol Immunol 246 (1999): 53-60. (Review)
PMID
10396039
Source
pubmed
Published In
Current topics in microbiology and immunology
Volume
246
Publish Date
1999
Start Page
53
End Page
60

Antigen drives very low affinity B cells to become plasmacytes and enter germinal centers.

In the first week of the primary immune response to the (4-hydroxy-3-nitrophenyl)acetyl (NP) hapten, plasmacytic foci and germinal centers (GCs) in C57BL/6 mice are comprised of polyclonal populations of B lymphocytes bearing the lambda1 L-chain (lambda1+). The Ig H-chains of these early populations of B cells are encoded by a variety of VH and D exons undiversified by hypermutation while later, oligoclonal populations are dominated by mutated rearrangements of the VH186.2 and DFL16.1 gene segments. To assess directly Ab affinities within these defined splenic microenvironments, representative VDJ rearrangements were recovered from B cells participating in the early immune response to NP, inserted into Ig H-chain expression cassettes, and transfected into J558L (H-; lambda1+) myeloma cells. These transfectoma Abs expressed a remarkably wide range of measured affinities (Ka = 5 x 10(4)-1.3 x 10(6) M(-1)) for NP. VDJs recovered from both foci and early GCs generated comparable affinities, suggesting that initial differentiation into these compartments occurs stochastically. We conclude that Ag normally activates B cells bearing an unexpectedly wide spectrum of Ab affinities and that this initial, promiscuous clonal activation is followed by affinity-driven competition to determine survival and clonal expansion within GCs and entry into the memory and bone marrow plasmacyte compartments.

Authors
Dal Porto, JM; Haberman, AM; Shlomchik, MJ; Kelsoe, G
MLA Citation
Dal Porto, JM, Haberman, AM, Shlomchik, MJ, and Kelsoe, G. "Antigen drives very low affinity B cells to become plasmacytes and enter germinal centers." J Immunol 161.10 (November 15, 1998): 5373-5381.
PMID
9820511
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
161
Issue
10
Publish Date
1998
Start Page
5373
End Page
5381

Predicted and inferred waiting times for key mutations in the germinal centre reaction: evidence for stochasticity in selection.

The germinal centre reaction (GCR) is a fundamental component of the immune response to T-dependent antigens, during which the immunoglobulin (Ig) genes of B cells experience somatic hypermutation and selection. A maximum-likelihood method on DNA sequence data from 16 individual germinal centres was used to infer that the waiting time for position 33 key (high-affinity) mutations in the anti-(4-hydroxy-3-nitrophenyl) acetyl (NP) response is 8.3 days. This is in marked contrast to the prediction of a key mutant each generation (waiting time about 1/3 day) obtained from a simple model and parameters available in the literature. This disagreement is resolved in part by the finding that the targeted base occurs in a cold spot for hypermutation, raising the predicted waiting time to 2.3 days, although this value remains significantly lower than that inferred from the sequence data. It is proposed that the remaining disparity is attributable to some further stochastic process in the GCR: many early key mutations arise but fail to 'take root' within the GC, either due to emigration or failure of cognate T cell/B cell interaction. Furthermore, it is argued that the frequency with which position 33 mutations are found in secondary responses to NP indicates the presence of selection after the GCR.

Authors
Radmacher, MD; Kelsoe, G; Kepler, TB
MLA Citation
Radmacher, MD, Kelsoe, G, and Kepler, TB. "Predicted and inferred waiting times for key mutations in the germinal centre reaction: evidence for stochasticity in selection." Immunol Cell Biol 76.4 (August 1998): 373-381.
PMID
9723780
Source
pubmed
Published In
Immunology and cell biology
Volume
76
Issue
4
Publish Date
1998
Start Page
373
End Page
381
DOI
10.1046/j.1440-1711.1998.00753.x

V(D)J hypermutation and DNA mismatch repair: vexed by fixation.

Authors
Kelsoe, G
MLA Citation
Kelsoe, G. "V(D)J hypermutation and DNA mismatch repair: vexed by fixation." Proc Natl Acad Sci U S A 95.12 (June 9, 1998): 6576-6577.
PMID
9618453
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
95
Issue
12
Publish Date
1998
Start Page
6576
End Page
6577

Dependence of germinal center B cells on expression of CD21/CD35 for survival.

Affinity-driven selection of B lymphocytes within germinal centers is critical for the development of high-affinity memory cells and host protection. To investigate the role of the CD21/CD35 coreceptor in B cell competition for follicular retention and survival within the germinal center, either Cr2+ or Cr2null lysozyme-specific transgenic B cells were adoptively transferred into normal mice immunized with duck (DEL) or turkey (TEL) lysozyme, which bind with different affinities. In mice injected with high-affinity turkey lysozyme, Cr2null B cells responded by follicular retention; however, they could not survive within germinal centers. This suggests that CD21 provides a signal independent of antigen that is required for survival of B cells in the germinal center.

Authors
Fischer, MB; Goerg, S; Shen, L; Prodeus, AP; Goodnow, CC; Kelsoe, G; Carroll, MC
MLA Citation
Fischer, MB, Goerg, S, Shen, L, Prodeus, AP, Goodnow, CC, Kelsoe, G, and Carroll, MC. "Dependence of germinal center B cells on expression of CD21/CD35 for survival." Science 280.5363 (April 24, 1998): 582-585.
PMID
9554848
Source
pubmed
Published In
Science
Volume
280
Issue
5363
Publish Date
1998
Start Page
582
End Page
585

Immunoglobulin gene hypermutation in germinal centers is independent of the RAG-1 V(D)J recombinase.

Antigen-driven somatic hypermutation in immunoglobulin genes coupled with stringent selection leads to affinity maturation in the B-lymphocyte populations present in germinal centers. To date, no gene(s) has been identified that drives the hypermutation process. The site-specific recombination of antigen-receptor gene segments in T and B lymphocytes is dependent on the expression of two recombination activating genes, RAG-1 and RAG-2. The RAG-1 and RAG-2 proteins are essential for the cleavage of DNA at highly conserved recombination signals to make double-strand breaks and their expression is sufficient to confer V(D)J recombination activity to non-lymphoid cells. Until very recently, expression of the V(D)J recombinase in adults was believed to be restricted to sites of primary lymphogenesis. However, several laboratories have now demonstrated expression of RAG-1 and RAG-2 and active V-to-(D)J recombination in germinal center B cells. This observation of active recombinase in germinal centers raises the issue of RAG-mediated nuclease activity as a component of V(D)J hypermutation. Here, we show that a transgenic kappa-light chain gene in a RAG-1-/- genetic background can acquire high frequencies of mutations. Thus, the RAG-1 protein is not essential for the machinery of immunoglobulin hypermutation. The genetic approaches to identifying the genes necessary for somatic hypermutation will require further studies on DNA-repair and immunodeficient models.

Authors
Zheng, B; Han, S; Spanopoulou, E; Kelsoe, G
MLA Citation
Zheng, B, Han, S, Spanopoulou, E, and Kelsoe, G. "Immunoglobulin gene hypermutation in germinal centers is independent of the RAG-1 V(D)J recombinase." Immunol Rev 162 (April 1998): 133-141. (Review)
PMID
9602359
Source
pubmed
Published In
Immunological Reviews
Volume
162
Publish Date
1998
Start Page
133
End Page
141

In situ studies of the primary immune response to (4-hydroxy-3-nitrophenyl)acetyl. V. Affinity maturation develops in two stages of clonal selection.

To examine the role of germinal centers (GCs) in the generation and selection of high affinity antibody-forming cells (AFCs), we have analyzed the average affinity of (4-hydroxy-3-nitrophenyl)acetyl (NP)-specific AFCs and serum antibodies both during and after the GC phase of the immune response. In addition, the genetics of NP-binding AFCs were followed to monitor the generation and selection of high affinity AFCs at the clonal level. NP-binding AFCs gradually accumulate in bone marrow (BM) after immunization and BM becomes the predominant locale of specific AFCs in the late primary response. Although the average affinity of NP-specific BM AFCs rapidly increased while GCs were present (GC phase), the affinity of both BM AFCs and serum antibodies continued to increase even after GCs waned (post-GC phase). Affinity maturation in the post-GC phase was also reflected in a shift in the distribution of somatic mutations as well as in the CDR3 sequences of BM AFC antibody heavy chain genes. Disruption of GCs by injection of antibody specific for CD154 (CD40 ligand) decreased the average affinity of subsequent BM AFCs, suggesting that GCs generate the precursors of high affinity BM AFCs; inhibition of CD154-dependent cellular interactions after the GC reaction was complete had no effect on high affinity BM AFCs. Interestingly, limited affinity maturation in the BM AFC compartment still occurs during the late primary response even after treatment with anti-CD154 antibody. Thus, GCs are necessary for the generation of high affinity AFC precursors but are not the only sites for the affinity-driven clonal selection responsible for the maturation of humoral immune responses.

Authors
Takahashi, Y; Dutta, PR; Cerasoli, DM; Kelsoe, G
MLA Citation
Takahashi, Y, Dutta, PR, Cerasoli, DM, and Kelsoe, G. "In situ studies of the primary immune response to (4-hydroxy-3-nitrophenyl)acetyl. V. Affinity maturation develops in two stages of clonal selection." J Exp Med 187.6 (March 16, 1998): 885-895.
Website
http://hdl.handle.net/10161/11493
PMID
9500791
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
187
Issue
6
Publish Date
1998
Start Page
885
End Page
895

Lymphocyte development and selection in germinal centers.

Authors
Przylepa, J; Himes, C; Kelsoe, G
MLA Citation
Przylepa, J, Himes, C, and Kelsoe, G. "Lymphocyte development and selection in germinal centers." Curr Top Microbiol Immunol 229 (1998): 85-104. (Review)
PMID
9479850
Source
pubmed
Published In
Current topics in microbiology and immunology
Volume
229
Publish Date
1998
Start Page
85
End Page
104

Immunosenescence and germinal center reaction.

Dysfunction of the immune system in aged individuals includes at least two important factors: accumulation of immunocytes with reduced function and accumulation of lymphocyte clones with self-reactive potential. Coincidently, there is a profound reduction of the germinal center reaction in the aged. While this reduction is likely the result of age-associated impairment in lymphocyte function (e.g. diminished response to costimulus, altered lymphokine production etc.), the reduction of germinal centers may itself make an important contribution to further immunological dysfunction.

Authors
Zheng, B; Han, S; Takahashi, Y; Kelsoe, G
MLA Citation
Zheng, B, Han, S, Takahashi, Y, and Kelsoe, G. "Immunosenescence and germinal center reaction." Immunol Rev 160 (December 1997): 63-77. (Review)
PMID
9476666
Source
pubmed
Published In
Immunological Reviews
Volume
160
Publish Date
1997
Start Page
63
End Page
77

V(D)J recombinase activity in a subset of germinal center B lymphocytes.

Reexpression of the V(D)J recombinase-activating genes RAG1 and RAG2 in germinal center B cells creates the potential for immunoglobulin gene rearrangement and the generation of new antigen receptor specificities. Intermediate products of V(D)J recombination are abundant in a subset of germinal center B cells, demonstrating that the kappa immunoglobulin light-chain locus becomes a substrate for renewed V(D)J recombinase activity. This recombinationally active cell compartment contains many heavy-chain VDJ rearrangements that encode low-affinity or nonfunctional antibody. In germinal centers, secondary V(D)J recombination may be induced by diminished binding to antigen ligands, thereby limiting abrupt changes in receptor specificity to B cells that are usually eliminated from the germinal center reaction. This restriction preserves efficient antigen-driven selection in germinal centers while allowing for saltations in the somatic evolution of B cells.

Authors
Han, S; Dillon, SR; Zheng, B; Shimoda, M; Schlissel, MS; Kelsoe, G
MLA Citation
Han, S, Dillon, SR, Zheng, B, Shimoda, M, Schlissel, MS, and Kelsoe, G. "V(D)J recombinase activity in a subset of germinal center B lymphocytes." Science 278.5336 (October 10, 1997): 301-305.
PMID
9323211
Source
pubmed
Published In
Science
Volume
278
Issue
5336
Publish Date
1997
Start Page
301
End Page
305

Distinctive characteristics of germinal center B cells.

A cardinal property of the immune system is its ability to respond to an antigen that was encountered years before with an accelerated and enhanced secondary response. The property of anamnestic reactions depends upon the formation of long-lived compartments of specialized T and B lymphocytes called memory cells. While the origin of the memory T-cell compartment is not known, germinal centers are the specialized sites for memory B-cell generation and the immunoglobulin V-region hypermutation necessary for the affinity maturation of serum antibody. Interestingly, the peripheral differentiation pathway that leads to this most mature B-cell state begins with the recapitulation of many characters of immature B lymphocytes in bone marrow. This review describes the distinctive cellular basis of germinal center reaction and the characteristics of B cells in germinal centers that later enter the memory pool.

Authors
Han, S; Zheng, B; Takahashi, Y; Kelsoe, G
MLA Citation
Han, S, Zheng, B, Takahashi, Y, and Kelsoe, G. "Distinctive characteristics of germinal center B cells." Semin Immunol 9.4 (August 1997): 255-260. (Review)
PMID
9237932
Source
pubmed
Published In
Seminars in Immunology
Volume
9
Issue
4
Publish Date
1997
Start Page
255
End Page
260
DOI
10.1006/smim.1997.0081

Neoteny in lymphocytes: Rag1 and Rag2 expression in germinal center B cells.

The products of the Rag1 and Rag2 genes drive genomic V(D)J rearrangements that assemble functional immunoglobulin and T cell antigen receptor genes. Expression of the Rag genes has been thought to be limited to developmentally immature lymphocyte populations that in normal adult animals are primarily restricted to the bone marrow and thymus. Abundant RAG1 and RAG2 protein and messenger RNA was detected in the activated B cells that populate murine splenic and Peyer's patch germinal centers. Germinal center B cells thus share fundamental characteristics of immature lymphocytes, raising the possibility that antigen-dependent secondary V(D)J rearrangements modify the peripheral antibody repertoire.

Authors
Han, S; Zheng, B; Schatz, DG; Spanopoulou, E; Kelsoe, G
MLA Citation
Han, S, Zheng, B, Schatz, DG, Spanopoulou, E, and Kelsoe, G. "Neoteny in lymphocytes: Rag1 and Rag2 expression in germinal center B cells." Science 274.5295 (December 20, 1996): 2094-2097.
PMID
8953043
Source
pubmed
Published In
Science
Volume
274
Issue
5295
Publish Date
1996
Start Page
2094
End Page
2097

Alternative pathways for the selection of antigen-specific peripheral T cells.

In the thymus, maturing lymphocytes receive activation signals mediated by the T-cell antigen receptor (TCR) that either promote clonal survival (positive selection) or induce apoptosis (negative selection). This balance between life and death is mirrored by the sensitivity of cortical thymocytes to apoptotic death induced by antibodies against the CD3 component of the TCR signal-transduction complex, bacterial superantigens that bind to the TCR beta-chain, and corticosteroids. In contrast, mature peripheral T cells are positively activated by anti-CD3 antibody or superantigens and are resistant to steroid-induced death. Here we show that in splenic germinal centres, T cells regain thymocyte-like sensitivity to TCR- and steroid-induced apoptosis and undergo antigen-driven positive and negative selection. T-cell responses elsewhere in the spleen are unaccompanied by programmed cell death. Our observations define a new differentiation pathway for peripheral T cells and suggest that germinal centres induce a lymphocyte phenotype necessary for the maintenance of self-tolerance.

Authors
Zheng, B; Han, S; Zhu, Q; Goldsby, R; Kelsoe, G
MLA Citation
Zheng, B, Han, S, Zhu, Q, Goldsby, R, and Kelsoe, G. "Alternative pathways for the selection of antigen-specific peripheral T cells." Nature 384.6606 (November 21, 1996): 263-266.
PMID
8918876
Source
pubmed
Published In
Nature
Volume
384
Issue
6606
Publish Date
1996
Start Page
263
End Page
266
DOI
10.1038/384263a0

Gamma delta T cell help of B cells is induced by repeated parasitic infection, in the absence of other T cells.

BACKGROUND: gamma delta T cells, like alpha beta T cells, are components of all well-studied vertebrate immune systems. Yet, the contribution of gamma delta T cells to immune responses is poorly characterized. In particular, it has not been resolved whether gamma delta cells, independent of any other T cells, can help B cells produce immunoglobulin and form germinal centers, anatomical foci of specialized T cell-B cell collaboration. RESULTS: TCR beta-/- mice, which lack all T cells except gamma delta T cells, routinely displayed higher levels of antibody than fully T cell-deficient mice. Repeated parasitic infection of TCR beta-/- mice, but not of T cell-deficient mice, increased antibody levels and induced germinal centers that contained B cells and monoclonal gamma delta cells in close juxtaposition. However, antibody specificities were more commonly against self than against the challenging pathogen. gamma delta T cell-B cell help was not induced by repeated inoculation of TCR beta-/- mice with mycobacterial antigens. CONCLUSIONS: In the absence of any other T cells, gamma delta T cell-B cell collaboration can be significantly enhanced by repeated infection. However, the lack of obvious enrichment for antibodies against the challenging pathogen distinguishes gamma delta T cell help from alpha beta T cell help induced under analogous circumstances. The increased production of generalized antibodies may be particularly relevant to the development of autoimmunity, which commonly occurs in patients suffering from alpha beta T cell deficiencies, such as AIDS.

Authors
Pao, W; Wen, L; Smith, AL; Gulbranson-Judge, A; Zheng, B; Kelsoe, G; MacLennan, IC; Owen, MJ; Hayday, AC
MLA Citation
Pao, W, Wen, L, Smith, AL, Gulbranson-Judge, A, Zheng, B, Kelsoe, G, MacLennan, IC, Owen, MJ, and Hayday, AC. "Gamma delta T cell help of B cells is induced by repeated parasitic infection, in the absence of other T cells." Curr Biol 6.10 (October 1, 1996): 1317-1325.
PMID
8939571
Source
pubmed
Published In
Current Biology
Volume
6
Issue
10
Publish Date
1996
Start Page
1317
End Page
1325

T helper cells in murine germinal centers are antigen-specific emigrants that downregulate Thy-1.

After immunization, activated splenic T cells proliferate in periarteriolar lymphoid sheaths (PALS) and subsequently migrate to the lymphoid follicle where they enter nascent germinal centers. Analysis of TCR V(D)J gene rearrangements indicates extensive emigration, frequently involving more than a single white pulp region. These migrants constitute a unique set of T helper cells that express antigen-specific alpha beta TCR, CD3, and CD4, but little or no Thy-1, a differentiation antigen present on the great majority of peripheral murine T lymphocytes. The origin of CD4+ Thy-1 follicular T cells appears to be the Thy+ population in the PALS, as both sets commonly share identical V(D)J rearrangements.

Authors
Zheng, B; Han, S; Kelsoe, G
MLA Citation
Zheng, B, Han, S, and Kelsoe, G. "T helper cells in murine germinal centers are antigen-specific emigrants that downregulate Thy-1." J Exp Med 184.3 (September 1, 1996): 1083-1091.
Website
http://hdl.handle.net/10161/11488
PMID
9064325
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
184
Issue
3
Publish Date
1996
Start Page
1083
End Page
1091

Regulation of the B cell response to T-dependent antigens by classical pathway complement.

Mice deficient in complement components C3 (C3 -/-) and C4 (C4 -/-) were found to have a profound defect in their Ab response to a T-dependent Ag (bacteriophage (phi X174). Characterization of the deficient mice demonstrated a diminished level of peanut agglutinin+ germinal centers and a failure in isotype switching despite normal B cell signaling in vitro. The nature of the defect was found to lie at the B cell level, as the T cells were primed in C3- and C4-deficient mice as well as those in wild-type mice. These results, and the finding that the defect could be partly reversed by a 10-fold increase in Ag dose, support the hypothesis that covalent attachment of complement ligands, i.e., C3b and C3d to the Ag-Ab complex, increases its immunogenicity.

Authors
Fischer, MB; Ma, M; Goerg, S; Zhou, X; Xia, J; Finco, O; Han, S; Kelsoe, G; Howard, RG; Rothstein, TL; Kremmer, E; Rosen, FS; Carroll, MC
MLA Citation
Fischer, MB, Ma, M, Goerg, S, Zhou, X, Xia, J, Finco, O, Han, S, Kelsoe, G, Howard, RG, Rothstein, TL, Kremmer, E, Rosen, FS, and Carroll, MC. "Regulation of the B cell response to T-dependent antigens by classical pathway complement." J Immunol 157.2 (July 15, 1996): 549-556.
PMID
8752901
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
157
Issue
2
Publish Date
1996
Start Page
549
End Page
556

CD28 is required for germinal center formation.

Previous studies have demonstrated that the T cell costimulatory molecule, CD28, is important in the development of humoral immunity. CD28-deficient mice exhibit defects in isotype switching and are more susceptible to pathogens that depend on an effective Ab response. To determine the basis of these defects, we have examined B cell responses of CD28-deficient mice at the microenvironmental level. Early in a normal T-dependent immune response, small numbers of B cells undergo activation in the T cell-rich zone of secondary lymphoid tissues and then migrate to B cell areas. These migrant B cells found developing germinal centers by proliferative expansion, during which individual cells acquire mutations in their rearranged Ig genes. B cell mutants retaining higher affinities for Ag undergo positive selection in germinal centers, resulting in the establishment of the memory B cell compartment. In the present study, we demonstrate that although potentially Ag-reactive cells within the lymphoid follicle accumulate following antigenic challenge, these cells fail to undergo proliferative expansion to form germinal centers and do not acquire somatic mutations in CD28-deficient animals. Thus, the CD28 activation pathway is required for Ab responses to T-dependent Ags. cell compartment. In the present study, we demonstrate that although potentially Ag-reactive cells within the lymphoid follicle accumulate following antigenic challenge, these cells fail to undergo proliferative expansion to form germinal centers and do not acquire somatic mutations in CD28-deficient animals. Thus, the CD28 activation pathway is required for Ab responses to T-dependent Ags.

Authors
Ferguson, SE; Han, S; Kelsoe, G; Thompson, CB
MLA Citation
Ferguson, SE, Han, S, Kelsoe, G, and Thompson, CB. "CD28 is required for germinal center formation." J Immunol 156.12 (June 15, 1996): 4576-4581.
PMID
8648099
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
156
Issue
12
Publish Date
1996
Start Page
4576
End Page
4581

The germinal center: a crucible for lymphocyte selection.

Antigen first activates T and B lymphocytes in the T-cell areas of secondary lymphoid tissues where cognate- and costimulus-dependent proliferation expands the population of reactive lymphocytes. Selected T- and B-cell progeny from this population migrate into B-cell zones to form germinal centers (GC), where intense proliferation, apoptosis, and V(D)J hypermutation takes place. It is now known that each of these processes occur in both compartments of GC lymphocytes and that the GC T-cell represents an unusual Thy-1- subset of alpha beta T-helper cells that may represent a terminally differentiated cell that is lost with the end of the GC reaction.

Authors
Kelsoe, G
MLA Citation
Kelsoe, G. "The germinal center: a crucible for lymphocyte selection." Semin Immunol 8.3 (June 1996): 179-184. (Review)
PMID
8738917
Source
pubmed
Published In
Seminars in Immunology
Volume
8
Issue
3
Publish Date
1996
Start Page
179
End Page
184
DOI
10.1006/smim.1996.0022

Germinal center formation, immunoglobulin class switching, and autoantibody production driven by "non alpha/beta" T cells.

The production of class-switched antibodies, particularly immunoglobulin (Ig) G1 and IgE, occurs efficiently in T cell receptor (TCR) alpha-/- mice that are congenitally devoid of alpha/beta T cells. This finding runs counter to a wealth of data indicating that IgG1 and IgE synthesis are largely dependent on the collaboration between B and alpha/beta T cells. Furthermore, many of the antibodies synthesized in TCR alpha-/- mice are reactive to a similar spectrum of self-antigens as that targeted by autoantibodies characterizing human systemic lupus erythematosus (SLE). SLE, too, is most commonly regarded as an alpha/beta T cell-mediated condition. To distinguish whether the development of autoantibodies in TCR alpha-/- mice is due to an intrinsic de-regulation of B cells, or to a heretofore poorly characterized collaboration between B and "non-alpha/beta T" cells, the phenotype has been reconstituted by transfer of various populations of B and non-alpha/beta T cells including cloned gamma/delta T cells derived from TCR alpha-/- mice, to severe combined immunodeficient (SCID) mice. The results establish that the reproducible production of IgG1 (including autoantibodies) is a product of non-alpha/beta T cell help that can be provided by gamma/delta T cells. This type of B-T collaboration sustains the production of germinal centers, lymphoid follicles that ordinarily are anatomical signatures of alpha/beta T-B cell collaboration. Thus, non-alpha/beta T cell help may drive Ig synthesis and autoreactivity under various circumstances, especially in cases of alpha/beta T cell immunodeficiency.

Authors
Wen, L; Pao, W; Wong, FS; Peng, Q; Craft, J; Zheng, B; Kelsoe, G; Dianda, L; Owen, MJ; Hayday, AC
MLA Citation
Wen, L, Pao, W, Wong, FS, Peng, Q, Craft, J, Zheng, B, Kelsoe, G, Dianda, L, Owen, MJ, and Hayday, AC. "Germinal center formation, immunoglobulin class switching, and autoantibody production driven by "non alpha/beta" T cells." J Exp Med 183.5 (May 1, 1996): 2271-2282.
PMID
8642336
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
183
Issue
5
Publish Date
1996
Start Page
2271
End Page
2282

Antibody response to a T-dependent antigen requires B cell expression of complement receptors.

Several lines of evidence indicate that antibody responses to T-dependent antigens require complement receptors expressed on either B lymphocytes or follicular dendritic cells. We have used RAG-2 deficient blastocyst complementation to create mice specifically lacking B cell complement receptors. Despite normal expression of complement receptor 1 (CR1[CD35]) and CR2 (CD21) on follicular dendritic cells, these mice have a profound defect in their capacity to mount a T-dependent antibody response. This is the first direct demonstration in vivo that B cell expression of complement receptors is required for a humoral immune response. This is the first direct demonstration in vivo that B cell expression of complement receptors is required for a humoral immune response. This suggests that CD21 and/or CD35 on B lymphocytes may be required for cellular activation, adsorptive endocytosis of antigen, recruitment to germinal centers, and/or protection from apoptosis during the humoral response to T-dependent antigens.

Authors
Croix, DA; Ahearn, JM; Rosengard, AM; Han, S; Kelsoe, G; Ma, M; Carroll, MC
MLA Citation
Croix, DA, Ahearn, JM, Rosengard, AM, Han, S, Kelsoe, G, Ma, M, and Carroll, MC. "Antibody response to a T-dependent antigen requires B cell expression of complement receptors." J Exp Med 183.4 (April 1, 1996): 1857-1864.
PMID
8666942
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
183
Issue
4
Publish Date
1996
Start Page
1857
End Page
1864

Life and death in germinal centers (redux).

Authors
Kelsoe, G
MLA Citation
Kelsoe, G. "Life and death in germinal centers (redux)." Immunity 4.2 (February 1996): 107-111. (Review)
PMID
8624801
Source
pubmed
Published In
Immunity
Volume
4
Issue
2
Publish Date
1996
Start Page
107
End Page
111

Somatic diversification of antibody responses.

Authors
Zheng, B; Kelsoe, G; Han, S
MLA Citation
Zheng, B, Kelsoe, G, and Han, S. "Somatic diversification of antibody responses." J Clin Immunol 16.1 (January 1996): 1-11. (Review)
PMID
8926280
Source
pubmed
Published In
Journal of Clinical Immunology
Volume
16
Issue
1
Publish Date
1996
Start Page
1
End Page
11

γδ T cell help of B cells is induced by repeated parasitic infection, in the absence of other T cells

Background: γδ T cells, like αβ T cells, are components of all well- studied vertebrate immune systems. Yet, the contribution of γδ T cells to immune responses is poorly characterized. In particular, it has not been resolved whether γδ cells, independent of any other T cells, can help B cells produce immunoglobulin and form germinal centers, anatomical foci of specialized T cell-B cell collaboration. Results: TCRβ(-/-) mice, which lack all T cells except γδ T cells, routinely displayed higher levels of antibody than fully T cell-deficient mice. Repeated parasitic infection of TCRβ(-/-) mice, but not of T cell-deficient mice, increased antibody levels and induced germinal centers that contained B cells and monoclonal γδ cells in close juxtaposition. However, antibody specificities were more commonly against self than against the challenging pathogen. γδ T cell-B cell help was not induced by repeated inoculation of TCRβ(-/-) mice with mycobacterial antigens. Conclusions: In the absence of any other T cells, γδ T cell-B cell collaboration can be significantly enhanced by repeated infection. However, the lack of obvious enrichment for antibodies against the challenging pathogen distinguishes γδ T cell help from αβ T cell help induced under analogous circumstances. The increased production of generalized antibodies may be particularly relevant to the development of autoimmunity, which commonly occurs in patients suffering from αβ T cell deficiencies, such as AIDS.

Authors
Pao, W; Wen, L; Smith, AL; Gulbranson-Judge, A; Zheng, B; Kelsoe, G; MacLennan, ICM; Owen, MJ; Hayday, AC
MLA Citation
Pao, W, Wen, L, Smith, AL, Gulbranson-Judge, A, Zheng, B, Kelsoe, G, MacLennan, ICM, Owen, MJ, and Hayday, AC. "γδ T cell help of B cells is induced by repeated parasitic infection, in the absence of other T cells." Current Biology 6.10 (1996): 1317-1325.
Source
scival
Published In
Current Biology
Volume
6
Issue
10
Publish Date
1996
Start Page
1317
End Page
1325

In situ studies of the primary immune response to (4-hydroxy-3-nitrophenyl)acetyl. IV. Affinity-dependent, antigen-driven B cell apoptosis in germinal centers as a mechanism for maintaining self-tolerance.

Germinal centers (GCs) are the sites of antigen-driven V(D)J gene hypermutation and selection necessary for the generation of high affinity memory B lymphocytes. Despite the antigen dependence of this reaction, injection of soluble antigen during an established primary immune response induces massive apoptotic death in GC B cells, but not in clonally related populations of nonfollicular B lymphoblasts and plasmacytes. Cell death in GCs occurs predominantly among light zone centrocytes, is antigen specific, and peaks within 4-8 h after injection. Antigen-induced programmed death does not involve cellular interactions mediated by CD40 ligand (CD40L) or Fas; disruption of GCs by antibody specific for CD40L was not driven by apoptosis and C57BL/6.lpr mice, though unable to express the Fas death trigger, remained fully susceptible to soluble antigen. Single injections of antigen did not significantly decrease GC numbers or average size, but repeated injections during an 18-h period resulted in fewer and substantially smaller GCs. As cell loss appeared most extensive in the light zone, decreased GC cellularity after prolonged exposure to soluble antigen implies that the Ig- centroblasts of the dark zone may require replenishment from light zone cells that have survived antigenic selection. GC cell death is avidity-dependent; oligovalent antigen induced relatively little apoptosis and GC B cells that survived long exposures to multivalent antigen expressed atypical VDJ rearrangements unlikely to encode high affinity antibody. Antigen-induced apoptotic death in GCs may represent a mechanism for the peripheral deletion of autoreactive B cell mutants much as the combinatorial repertoire of immature B lymphocytes is censored in the bone marrow.

Authors
Han, S; Zheng, B; Dal Porto, J; Kelsoe, G
MLA Citation
Han, S, Zheng, B, Dal Porto, J, and Kelsoe, G. "In situ studies of the primary immune response to (4-hydroxy-3-nitrophenyl)acetyl. IV. Affinity-dependent, antigen-driven B cell apoptosis in germinal centers as a mechanism for maintaining self-tolerance." J Exp Med 182.6 (December 1, 1995): 1635-1644.
Website
http://hdl.handle.net/10161/11487
PMID
7500008
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
182
Issue
6
Publish Date
1995
Start Page
1635
End Page
1644

Ig VH hypermutation is absent in the germinal centers of aged mice.

After injection with immunogenic conjugates of the hapten (4-hydroxy-3-nitrophenyl)acetyl (NP), two distinct B cell populations can be identified in the spleen during the primary response. One of these populations is specialized for Ab production; the other, the germinal centers (GCs), has been identified as the site of Ig somatic hypermutation. Ag-driven selection of GC B cells bearing mutated receptors with higher affinity leads to the affinity maturation of serum Ab and increased protective humoral immunity. Microdissection of GC B cell populations specific for NP and sequencing of the recovered Ig heavy chain variable region genes revealed that the somatic hypermutation process is absent in the GCs of aged C57BL/6 mice. However, selection for Ag appears to occur in the absence of hypermutation in the form of competition between unmutated clones of Ag-activated B lymphocytes. Thus, affinity maturation in these animals is limited to the affinities of Ab encoded by the germline.

Authors
Miller, C; Kelsoe, G
MLA Citation
Miller, C, and Kelsoe, G. "Ig VH hypermutation is absent in the germinal centers of aged mice." J Immunol 155.7 (October 1, 1995): 3377-3384.
PMID
7561032
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
155
Issue
7
Publish Date
1995
Start Page
3377
End Page
3384

Cellular interaction in germinal centers. Roles of CD40 ligand and B7-2 in established germinal centers.

Costimulatory interactions between T and B lymphocytes are crucial for T cell activation and B cell proliferation and differentiation. We have compared the roles of CD40L and B7-2 in the initiation and maturation of humoral immunity by administering anti-CD40 ligand (L) or anti-B7-2 Ab during the early (days -1 to 3) or late (days 6-10) phases of primary responses to thymus-dependent (Td) and -independent (Ti) Ags. Germinal center (GC) formation in response to a Td Ag was inhibited completely by the early administration of anti-CD40L or anti-B7-2 Abs. Later in the response, established GCs remained sensitive to anti-CD40L but were resistant to treatment with anti-B7-2. However, Ig hypermutation was reduced dramatically in GCs of anti-B7-2-treated mice and humoral memory was impaired. Early administration of anti-CD40L reduced serum Ab levels to approximately 10% of controls, whereas early treatment with anti-B7-2 reduced Ab production by only 50%. Later treatments with either Ab had no effect on Ab production. Response to a type II Ti Ag was more resistant than Td responses to interruption of costimulatory interactions. Our findings suggest that the costimulatory roles of CD40:CD40L and B7-2:CD28/CTLA-4 differ in the GC; administration of anti-CD40L abrogates an established GC reaction, whereas Ab to B7-2 suppresses Ig hypermutation and entry into the B cell memory compartment. Once B cells have entered the differentiation pathway to Ab production, neither CD40L nor B7-2 is necessary for their continued differentiation and persistence.

Authors
Han, S; Hathcock, K; Zheng, B; Kepler, TB; Hodes, R; Kelsoe, G
MLA Citation
Han, S, Hathcock, K, Zheng, B, Kepler, TB, Hodes, R, and Kelsoe, G. "Cellular interaction in germinal centers. Roles of CD40 ligand and B7-2 in established germinal centers." J Immunol 155.2 (July 15, 1995): 556-567.
PMID
7541819
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
155
Issue
2
Publish Date
1995
Start Page
556
End Page
567

The germinal center reaction.

Authors
Kelsoe, G
MLA Citation
Kelsoe, G. "The germinal center reaction." Immunol Today 16.7 (July 1995): 324-326. (Review)
PMID
7576065
Source
pubmed
Published In
Immunology Today
Volume
16
Issue
7
Publish Date
1995
Start Page
324
End Page
326

Facultative role of germinal centers and T cells in the somatic diversification of IgVH genes.

The development of memory B cells takes place in germinal centers (GC) of lymphoid follicles where antigen-driven lymphocytes undergo somatic hypermutation and affinity selection, presumably under the influence of helper T cells. However, the mechanisms that drive this complex response are not well understood. We explored the relationship between GC formation and the onset of hypermutation in response to the hapten phosphorylcholine (PC) coupled to antigenic proteins in mice bearing different frequencies of CD4+ T cells. PC-reactive GC were identified by staining frozen splenic sections with peanut agglutinin (PNA) and with monoclonal Abs against AB1-2, a dominant idiotope of T15+ anti-PC antibody. The nucleotide sequences of rearranged T15 VH1 genes were determined from polymerase chain reaction amplifications of genomic DNA from microdissected GC B cells. T15+ GC became fully developed by day 6-7 after primary immunization of euthymic mice with either PC-keyhole limpet hemocyanin (KLH) or PC-chicken gamma globulin (CGG). Yet the VH1 gene segments recovered from the primary GC as late as day 10-14 had low numbers of mutations, in contrast to responses to the haptens nitrophenyl or oxazolone that sustain high levels of hypermutation after GC formation. PC-reactive B cells proliferate in histologically typical GC for considerable periods with no or little somatic hypermutation; the signals for GC formation are independent of those for the activation of hypermutation. We then examined GC 7 d after secondary immunization with PC-KLH in euthymic mice, in nu/nu mice reconstituted with limited numbers of normal CD4+ cells before priming (CD4(+)-nu/nu) and in nu/nu mice. All of these animals develop T15+ GC after antigen priming, however, the patterns of V gene mutations in the secondary GC reflected the levels of CD4+ cells present during the primary response. VDJ sequences from secondary GC of euthymic mice were heavily mutated, but most of these mutations were shared among all related (identical VDJ joints) sequences suggesting the proliferation of mutated, memory B cells, with little de novo somatic hypermutation. In contrast, the patterns of V gene diversity in secondary GC from CD4(+)-nu/nu mice suggested that there was ongoing mutation and clonal diversification during the first week after rechallenge. The secondary GC from T cell-deficient, nu/nu mice showed little evidence for mutational and/or recombinational diversity of T15+ B cells. We conclude that the participation of CD4+ helper cells is required for full activation of the mutator in GC and takes place in a dose-dependent fashion.

Authors
Miller, C; Stedra, J; Kelsoe, G; Cerny, J
MLA Citation
Miller, C, Stedra, J, Kelsoe, G, and Cerny, J. "Facultative role of germinal centers and T cells in the somatic diversification of IgVH genes." J Exp Med 181.4 (April 1, 1995): 1319-1331.
PMID
7535332
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
181
Issue
4
Publish Date
1995
Start Page
1319
End Page
1331

The nude mutation results in impaired primary antibody repertoire.

We have studied the effect of the nude mutation and/or T lymphocytes on the development of V gene germ-line repertoire in neonatal athymic (nu/nu) and euthymic (+/nu) littermates. A total of 2.35 x 10(6) and 1.47 x 10(6) B lymphocyte clones from nu/nu and +/nu neonates, respectively, were examined for the expression of select VH (J558, J606, S107, 36-60, 7183 and Q52) and Vx (1, 2, 8 and 9) gene families as well as VH (J558, S107) + Vx (1, 9) associations. Data showed that the nude mutation, whether homozygous or heterozygous, significantly affects VH and Vx gene expression as well as VH and Vx pairings and, thus, provide evidence for a defective development of B cell repertoire in both athymic nude (nu/nu) and euthymic (+/nu) mice. In addition, an analysis of 3.34 x 10(6) B lymphocyte clones from adult C57BL/6 mice showed non-stochastic association between VHJ558 + Vx1 gene families that suggests restrictions on clonal population in order to maintain homeostasis in the immune system. Studies outlined here, therefore, describe an hitherto unknown defect in the development of B lymphocyte repertoire as a result of the nude mutation which is independent of thymic dysgenesis.

Authors
Kaushik, A; Kelsoe, G; Jaton, JC
MLA Citation
Kaushik, A, Kelsoe, G, and Jaton, JC. "The nude mutation results in impaired primary antibody repertoire." Eur J Immunol 25.2 (February 1995): 631-634.
PMID
7875225
Source
pubmed
Published In
European Journal of Immunology
Volume
25
Issue
2
Publish Date
1995
Start Page
631
End Page
634
DOI
10.1002/eji.1830250249

In situ studies of the germinal center reaction.

Authors
Kelsoe, G
MLA Citation
Kelsoe, G. "In situ studies of the germinal center reaction." Adv Immunol 60 (1995): 267-288. (Review)
PMID
8607371
Source
pubmed
Published In
Advances in immunology
Volume
60
Publish Date
1995
Start Page
267
End Page
288

Skewed VH and V kappa gene family expression and pairing occurs among B lymphocytes in autoimmune motheaten mice.

Motheaten mice homozygous for the recessive mev mutation develop a fatal immunodeficiency syndrome associated with hypergammaglobulinemia, thymic aplasia, production of autoantibodies and development of a severe lupus like systemic autoimmune disease. Most B lymphocytes in this mutant strain belong to B-1 subset. We have addressed the question if differences existed in the V-gene repertoire of autoimmune mev/mev mice as compared to phenotypically normal mev/+ and C57BL/6 background strain by examining the VH and V kappa gene family expression as well as the association of VH and V kappa gene families among B lymphocyte clones. The data outlined here demonstrate that both the expression of VH and V kappa gene families and their association is skewed in mev/mev mice, suffering from systemic autoimmune disease, and differs significantly from phenotypically normal mev/+ litter mates as well as the C57BL/6 background strain. In addition, VH+V kappa gene family pairs in phenotypically normal mev/+ differed from normal C57BL/6 mice suggesting that motheaten mutation, whether homozygous or heterozygous, alters the development of the B lymphocyte repertoire. These observations suggest positive selection of B-1 lymphocytes in autoimmune motheaten mice either as a result of selective processes, via receptor-ligand interactions, operating on the development of the primary antibody repertoire or defective B lymphocyte haematopoiesis due to the deficiency of haematopoietic cell phosphatase involved in determining the threshold by which B cells respond to self antigen(s).

Authors
Saitoh, Y; Kelsoe, G; Bona, C; Kaushik, A
MLA Citation
Saitoh, Y, Kelsoe, G, Bona, C, and Kaushik, A. "Skewed VH and V kappa gene family expression and pairing occurs among B lymphocytes in autoimmune motheaten mice." Autoimmunity 21.3 (1995): 185-193.
PMID
8822276
Source
pubmed
Published In
Autoimmunity (Informa)
Volume
21
Issue
3
Publish Date
1995
Start Page
185
End Page
193

Hypermutation in T cells questioned [5]

Authors
Bachl, J; Wabl, M; Kelsoe, G; Zheng, B; Kepler, TB
MLA Citation
Bachl, J, Wabl, M, Kelsoe, G, Zheng, B, and Kepler, TB. "Hypermutation in T cells questioned [5]." Nature 375.6529 (1995): 285-286.
PMID
7753192
Source
scival
Published In
Nature
Volume
375
Issue
6529
Publish Date
1995
Start Page
285
End Page
286

Locus-specific somatic hypermutation in germinal centre T cells.

Somatic hypermutation and affinity-driven selection of active immunoglobulin genes occur in germinal centres (GCs), resulting in the generation of high-affinity memory B cells. In contrast, T lymphocytes do not require the germinal centre microenvironment to establish memory and the T-cell antigen receptor (TCR) genes, though homologous to immunoglobulin genes, are believed to be incapable of hypermutation. Here we present direct evidence that the small population of antigen-specific T cells that are recruited into splenic GCs acquire mutations in the variable region of genes encoding TCR alpha-chains (V alpha) but not those of beta-chains. These locus-specific mutations reach frequencies comparable to mutated immunoglobulin VH exons recovered from the same site and exhibit similar substitution biases and DNA strand polarity. T cells bearing identical mutations appear in multiple GCs, raising the possibility that some cells bearing mutant TCRs may re-enter the peripheral lymphocyte pool.

Authors
Zheng, B; Xue, W; Kelsoe, G
MLA Citation
Zheng, B, Xue, W, and Kelsoe, G. "Locus-specific somatic hypermutation in germinal centre T cells." Nature 372.6506 (December 8, 1994): 556-559.
PMID
7990929
Source
pubmed
Published In
Nature
Volume
372
Issue
6506
Publish Date
1994
Start Page
556
End Page
559
DOI
10.1038/372556a0

B cell diversification and differentiation in the periphery.

Authors
Kelsoe, G
MLA Citation
Kelsoe, G. "B cell diversification and differentiation in the periphery." J Exp Med 180.1 (July 1, 1994): 5-6.
PMID
8006600
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
180
Issue
1
Publish Date
1994
Start Page
5
End Page
6

Lack of B7-2 expression in the germinal centers of aged mice

The humoral immune response is known to be depressed in the aged mouse. This immunodeficiency is associated with an impairment in the formation of germinal centers, the anatomic site of antibody affinity maturation and memory B cell generation. The formation of germinal centers is dependent on the expression of costimulatory molecules. We have investigated the expression of the recently identified costimulatory molecule, B7-2, and another activation marker, GL7, in the germinal centers of aged mice. In contrast to young syngeneic controls, B7-2 and GL7 are not expressed in the germinal centers of aged C57BL/6 mice. Administration of an antibody specific for the B7-2 molecule produces a defective immune response in young mice that is similar to that seen in old animals. Thus, we propose that the absence of differentiation pathways mediated by B7-2 in aged mice defines a major, age-related immunodeficiency.

Authors
Miller, C; Kelsoe, G; Han, S
MLA Citation
Miller, C, Kelsoe, G, and Han, S. "Lack of B7-2 expression in the germinal centers of aged mice." Aging: Immunology and Infectious Disease 5.3-4 (1994): 249-257.
Source
scival
Published In
Aging: Immunology and Infectious Disease
Volume
5
Issue
3-4
Publish Date
1994
Start Page
249
End Page
257

In situ studies of the primary immune response to (4-hydroxy-3-nitrophenyl)acetyl. III. The kinetics of V region mutation and selection in germinal center B cells.

In the murine spleen, germinal centers are the anatomic sites for antigen-driven hypermutation and selection of immunoglobulin (Ig) genes. To detail the kinetics of Ig mutation and selection, 178 VDJ sequences from 16 antigen-induced germinal centers were analyzed. Although germinal centers appeared by day 4, mutation was not observed in germinal center B cells until day 8 postimmunization; thereafter, point mutations favoring asymmetrical transversions accumulated until day 14. During this period, strong phenotypic selection on the mutant B lymphocytes was inferred from progressively biased distributions of mutations within the Ig variable region, the loss of crippling mutations, decreased relative clonal diversity, and increasingly restricted use of canonical gene segments. The period of most intense selection on germinal center B cell populations preceded significant levels of mutation and may represent a physiologically determined restriction on B cells permitted to enter the memory pathway. Noncanonical Ig genes recovered from germinal centers were mostly unmutated although they probably came from antigen-reactive cells. Together, these observations demonstrate that the germinal center microenvironment is rich and temporally complex but may not be constitutive for somatic hypermutation.

Authors
Jacob, J; Przylepa, J; Miller, C; Kelsoe, G
MLA Citation
Jacob, J, Przylepa, J, Miller, C, and Kelsoe, G. "In situ studies of the primary immune response to (4-hydroxy-3-nitrophenyl)acetyl. III. The kinetics of V region mutation and selection in germinal center B cells." J Exp Med 178.4 (October 1, 1993): 1293-1307.
Website
http://hdl.handle.net/10161/11486
PMID
8376935
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
178
Issue
4
Publish Date
1993
Start Page
1293
End Page
1307

Pairing of VH gene families with the lambda 1 light chain: evidence for a non-stochastic association.

The frequencies at which four VH gene families pair with the lambda 1 light (L) chain were determined by sequential hybridization of VH- and lambda 1-specific DNA probes to mitogen-induced colonies of B cells. Analysis of pair frequencies indicates that the repertoire of lambda L chain antibodies is generated by the stochastic pairing of smaller 3'-to-mid-locus VH gene families (X-24, S107, Q52). However, the large 5' VH J558 family appeared to associate with the lambda 1 L chain non-stochastically; the frequency of VHJ558/lambda 1+ colonies among all lambda 1+ colonies was significantly lower than the frequency of J558 expression among all (C mu+) B cell colonies. This difference suggests that selection, either intrinsic at the level of rearrangement or heavy and L chain pairing, or extrinsic following surface immunoglobulin expression, may operate to shape the lambda antibody repertoire prior to the introduction of exogenous antigen.

Authors
Yurovsky, VV; Kelsoe, G
MLA Citation
Yurovsky, VV, and Kelsoe, G. "Pairing of VH gene families with the lambda 1 light chain: evidence for a non-stochastic association." Eur J Immunol 23.8 (August 1993): 1975-1979.
PMID
8344362
Source
pubmed
Published In
European Journal of Immunology
Volume
23
Issue
8
Publish Date
1993
Start Page
1975
End Page
1979
DOI
10.1002/eji.1830230837

Sites of B-cell activation in vivo.

Novel techniques have made possible in situ analyses of the lymphocyte populations responding to antigen. In the spleen, antigen-specific T and B cells are first observed in the periarteriolar lymphoid sheath. Following conjugate formation between specific T and B lymphocytes, B-cell proliferation and differentiation takes place in two distinct sites, the periarteriolar lymphoid sheath-associated foci and germinal centers.

Authors
Kelsoe, G; Zheng, B
MLA Citation
Kelsoe, G, and Zheng, B. "Sites of B-cell activation in vivo." Curr Opin Immunol 5.3 (June 1993): 418-422. (Review)
PMID
8347301
Source
pubmed
Published In
Current Opinion in Immunology
Volume
5
Issue
3
Publish Date
1993
Start Page
418
End Page
422

In situ studies of the primary immune response to (4-hydroxy-3-nitrophenyl)acetyl. II. A common clonal origin for periarteriolar lymphoid sheath-associated foci and germinal centers.

In the genetically restricted response that follows immunization with (4-hydroxy-3-nitrophenyl)acetyl coupled to protein carriers, two distinct populations of B cells are observed in the spleens of C57BL/6 mice. By 48 h postimmunization, foci of antigen-binding B cells appear along the periphery of the periarteriolar lymphoid sheaths. These foci expand to contain large numbers of antibody-forming cells that neither bind the lectin, peanut agglutinin, nor mutate the rearranged immunoglobulin variable region loci. Germinal centers containing peanut agglutinin-positive B cells can be observed by 96-120 h after immunization. Although specific for the immunizing hapten, these B cells do not produce substantial amounts of antibody, but are the population that undergoes somatic hypermutation and affinity-driven selection. Both focus and germinal center populations are pauciclonal, founded, on average, by three or fewer B lymphocytes. Despite the highly specialized roles of the focus (early antibody production) and germinal center (higher affinity memory cells) B cell populations, analysis of VH to D to JH joins in neighboring foci and germinal centers demonstrate that these B cell populations have a common clonal origin.

Authors
Jacob, J; Kelsoe, G
MLA Citation
Jacob, J, and Kelsoe, G. "In situ studies of the primary immune response to (4-hydroxy-3-nitrophenyl)acetyl. II. A common clonal origin for periarteriolar lymphoid sheath-associated foci and germinal centers." J Exp Med 176.3 (September 1, 1992): 679-687.
Website
http://hdl.handle.net/10161/11492
PMID
1512536
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
176
Issue
3
Publish Date
1992
Start Page
679
End Page
687

Aging and humoral immunity.

If human antibody responses undergo molecular shifts similar to those identified in mice, the appropriate immunization strategy for the elderly would be a passive administration of the protective antibody from young donors rather than an attempt to boost the individual's own response with a more potent vaccine, because the shifted immune system can no longer make the right kind of antibody.

Authors
Goidl, EA; Cerny, J; Kelsoe, G; Schulze, DH
MLA Citation
Goidl, EA, Cerny, J, Kelsoe, G, and Schulze, DH. "Aging and humoral immunity." Md Med J 41.7 (July 1992): 609-613.
PMID
1640817
Source
pubmed
Published In
Maryland Medicine
Volume
41
Issue
7
Publish Date
1992
Start Page
609
End Page
613

In situ studies of the antigen-driven somatic hypermutation of immunoglobulin genes.

Authors
Jacob, J; Miller, C; Kelsoe, G
MLA Citation
Jacob, J, Miller, C, and Kelsoe, G. "In situ studies of the antigen-driven somatic hypermutation of immunoglobulin genes." Immunol Cell Biol 70 ( Pt 2) (April 1992): 145-152. (Review)
PMID
1398774
Source
pubmed
Published In
Immunology and cell biology
Volume
70 ( Pt 2)
Publish Date
1992
Start Page
145
End Page
152
DOI
10.1038/icb.1992.19

Intraclonal generation of antibody mutants in germinal centres.

The generation and selection of somatic antibody mutants are key elements of acquired immunity, essential for the affinity maturation of antibody responses dependent on T cells. The mutants are generated through a mechanism that introduces point mutations at high rate into rearranged variable (V) region genes in the course of cell proliferation. Their appearance coincides with the generation of germinal centres, which are characterized by oligoclonal B-cell proliferation and have been suggested to be the microenvironment in which antibody mutants are generated. We report here direct evidence for this hypothesis. Rearranged V-region genes were amplified from the genomic DNA of cells picked from individual germinal centres. The sequence analysis of these genes revealed that most represent cells of distinct B-cell clones which expanded locally, generating somatic antibody mutants at high rate. By contrast, antigen-induced proliferation of B cells at another site, periarteriolar lymphocyte sheath-associated foci, was not associated with somatic hypermutation.

Authors
Jacob, J; Kelsoe, G; Rajewsky, K; Weiss, U
MLA Citation
Jacob, J, Kelsoe, G, Rajewsky, K, and Weiss, U. "Intraclonal generation of antibody mutants in germinal centres." Nature 354.6352 (December 5, 1991): 389-392.
PMID
1956400
Source
pubmed
Published In
Nature
Volume
354
Issue
6352
Publish Date
1991
Start Page
389
End Page
392
DOI
10.1038/354389a0

In situ studies of the primary immune response to (4-hydroxy-3-nitrophenyl)acetyl. I. The architecture and dynamics of responding cell populations.

After primary immunization with an immunogenic conjugate of (4-hydroxy-3-nitrophenyl)acetyl, two anatomically and phenotypically distinct populations of antibody-forming cells arise in the spleen. As early as 2 d after immunization, foci of antigen-binding B cells are observed along the periphery of the periarteriolar lymphoid sheaths. These foci expand, occupying as much as 1% of the splenic volume by day 8 of the response. Later, foci grow smaller and are virtually absent from the spleen by day 14. A second responding population, germinal center B cells, appear on day 8-10 and persist at least until day 16 post-immunization. Individual foci and germinal centers represent discrete pauciclonal populations that apparently undergo somatic evolution in the course of the primary response. We suggest that foci may represent regions of predominantly interclonal competition for antigen among unmutated B cells, while germinal centers are sites of intraclonal clonal competition between mutated sister lymphocytes.

Authors
Jacob, J; Kassir, R; Kelsoe, G
MLA Citation
Jacob, J, Kassir, R, and Kelsoe, G. "In situ studies of the primary immune response to (4-hydroxy-3-nitrophenyl)acetyl. I. The architecture and dynamics of responding cell populations." J Exp Med 173.5 (May 1, 1991): 1165-1175.
Website
http://hdl.handle.net/10161/11490
PMID
1902502
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
173
Issue
5
Publish Date
1991
Start Page
1165
End Page
1175

Contribution of the VH11 gene family to mitogen-responsive B cell repertoire in C57BL/6 mice.

The contribution of VH11 gene family to the development of the primary B cell repertoire has been studied by analyzing 1.8 x 10(4) mitogen induced B lymphocyte colonies. The data demonstrate that VH11 family is predominantly expressed among neonatal splenic as well as adult peritoneal B cell colonies, both rich in Ly-1+ B cells. VH11 gene family expression among B splenocytes decreases during ontogeny and VH11 family pairs stochastically with different V kappa families among mitogen-activated neonatal B cell colonies, which are representative of an antigen unselected B cell repertoire. Thus, an increased VH11 expression among peritoneal and neonatal B cells points towards its biased expression among Ly-1+ B lymphocytes. The restricted V gene rearrangements and VH11-V kappa 9 pairing observed among anti-bromelain-treated mouse red blood cells autoantibodies are likely to be an outcome of both intrinsic gene recombination processes per se as well as selection by an autoantigen and/or local selective environmental factors.

Authors
Kaushik, A; Reininger, L; Kelsoe, G; Jaton, JC; Bona, C
MLA Citation
Kaushik, A, Reininger, L, Kelsoe, G, Jaton, JC, and Bona, C. "Contribution of the VH11 gene family to mitogen-responsive B cell repertoire in C57BL/6 mice." Eur J Immunol 21.3 (March 1991): 827-830.
PMID
1901267
Source
pubmed
Published In
European Journal of Immunology
Volume
21
Issue
3
Publish Date
1991
Start Page
827
End Page
830
DOI
10.1002/eji.1830210344

Cloning of murine splenic T lymphocytes and natural killer (NK) cells on filter paper discs: detection of a novel NK/T phenotype.

Discrete colonies of splenocytes were grown on filter paper discs in the presence of concanavalin A and interleukin 2. Phenotypic analysis of the colonies indicated that the majority expressed the Thy-1.2 marker and 72% of these co-expressed the CD3 molecule. Of the colonies 20%-25% were NK 1.1+ and they developed regardless of the presence of Con A in the culture medium, a property of the NK lineage. In addition, Thy-1.2+ colonies developed when splenocytes from scid mice, which lack mature T and B cells, were grown both in the presence and absence of concanavalin A. These results demonstrate that colonies of murine splenic T lymphocytes and NK cells could be successfully grown on filter paper discs and phenotypically characterized. With this colonies technique, it was possible to identify a novel subset of NK 1.1+ colonies that co-expresses CD3 and shares growth properties with T cell colonies.

Authors
Sarzotti, M; Kumar, V; Bennett, M; Kelsoe, G
MLA Citation
Sarzotti, M, Kumar, V, Bennett, M, and Kelsoe, G. "Cloning of murine splenic T lymphocytes and natural killer (NK) cells on filter paper discs: detection of a novel NK/T phenotype." Eur J Immunol 21.3 (March 1991): 635-641.
PMID
1672644
Source
pubmed
Published In
European Journal of Immunology
Volume
21
Issue
3
Publish Date
1991
Start Page
635
End Page
641
DOI
10.1002/eji.1830210315

The primary antibody repertoire: the somatic genotype and paratopic phenotype of B-cell populations.

Authors
Kelsoe, G
MLA Citation
Kelsoe, G. "The primary antibody repertoire: the somatic genotype and paratopic phenotype of B-cell populations." Immunol Ser 55 (1991): 81-92. (Review)
PMID
1954293
Source
pubmed
Published In
Immunology series
Volume
55
Publish Date
1991
Start Page
81
End Page
92

Pairing of VK and VK gene families in self-reactive antibodies.

Authors
Bona, CA; Saitoh, Y; Kelsoe, G
MLA Citation
Bona, CA, Saitoh, Y, and Kelsoe, G. "Pairing of VK and VK gene families in self-reactive antibodies." J Clin Immunol 10.5 (September 1990): 223-236. (Review)
PMID
2266149
Source
pubmed
Published In
Journal of Clinical Immunology
Volume
10
Issue
5
Publish Date
1990
Start Page
223
End Page
236

Stochastic pairing of heavy-chain and kappa light-chain variable gene families occurs in polyclonally activated B cells.

Frequencies of 25 immunoglobulin heavy-chain and kappa light-chain variable (VH + V kappa) gene-family pairings expressed in splenic B-cell populations were determined by hybridization of VH- and V kappa-family-specific DNA probes to mitogen-induced B-cell colonies from C57BL/6 mice or hybridomas derived from BALB/c and NZB mice. Both analyses support the conclusion that VH and V kappa gene families pair without bias; as would be expected for random association, the frequencies of specific VH + V kappa pairs may be estimated by the product of the independent VH and V kappa frequencies. Based upon the frequencies at which 9 VH and 12 V kappa gene families are expressed, we calculated the expected usage for approximately 100 VH + V kappa family pairings in neonatal and adult C57BL/6 mice. Variability in the expression of such VH + V kappa pairings is considerable; pairs representing greater than 10% to less than 0.01% of the splenic B-cell population occur. This variability is most pronounced in the neonate, where 6 VH + V kappa family pairs account for nearly 40% of all mitogen-reactive B cells. As the neonate matures, the distribution of frequencies for VH + V kappa pairings becomes more nearly uniform. This process may underlie the patterned acquisition of humoral immune responsiveness.

Authors
Kaushik, A; Schulze, DH; Bonilla, FA; Bona, C; Kelsoe, G
MLA Citation
Kaushik, A, Schulze, DH, Bonilla, FA, Bona, C, and Kelsoe, G. "Stochastic pairing of heavy-chain and kappa light-chain variable gene families occurs in polyclonally activated B cells." Proc Natl Acad Sci U S A 87.13 (July 1990): 4932-4936.
PMID
2114644
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
87
Issue
13
Publish Date
1990
Start Page
4932
End Page
4936

Antigen-binding repertoire and Ig H chain gene usage among B cell hybridomas from normal and autoimmune mice.

LPS-stimulated B cells were used to generate a panel of mAb that were a random sample of the preimmune repertoire of C57BL/6 and highly autoimmune, viable motheaten mice. These mAb were tested for reactivity to a number of "self" and foreign Ag. Binding that could be detected only at nM mAb concentrations or less was considered significant. We found that a surprisingly high number of the mAb bound one or more of the Ag tested, and many mAb bound more than a single Ag. Ag-induced mAb were likewise tested and found to have greatly reduced cross-reactivities. We found no significant differences, either in frequency of Ag binding or degree of cross-reactivity, between normal and autoimmune mice. Furthermore, the frequency with which a given Ag was bound by our panel of mAb was found to be proportional to the size of the Ag. The frequency with which individual VH gene families were expressed by our panel was consistent with a stochastic usage of VH genes in the preimmune repertoire. We interpret these data as showing that the preimmune repertoire is highly cross-reactive and that the activation of autoreactive clones in autoimmune animals is due to a defect in cellular regulation rather than a difference in repertoire.

Authors
Striebich, CC; Miceli, RM; Schulze, DH; Kelsoe, G; Cerny, J
MLA Citation
Striebich, CC, Miceli, RM, Schulze, DH, Kelsoe, G, and Cerny, J. "Antigen-binding repertoire and Ig H chain gene usage among B cell hybridomas from normal and autoimmune mice." J Immunol 144.5 (March 1, 1990): 1857-1865.
PMID
2106554
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
144
Issue
5
Publish Date
1990
Start Page
1857
End Page
1865

Murine V kappa gene expression does not follow the VH paradigm.

V kappa gene family expression among LPS-reactive murine B lymphocytes, unlike that of VH gene families, is not proportional to genomic complexity, i.e., nonstoichiometric. Furthermore, no positional bias for the overexpression of J-proximal V kappa genes (V kappa 21) is observed among neonatal B lymphocytes. Yet, the V kappa 1 and V kappa 9 families located in the center of V kappa locus are preferentially used by neonatal B splenocytes. Thus, the mechanisms of V kappa gene rearrangement and expression appear to differ significantly from those controlling the VH locus.

Authors
Kaushik, A; Schulze, DH; Bona, C; Kelsoe, G
MLA Citation
Kaushik, A, Schulze, DH, Bona, C, and Kelsoe, G. "Murine V kappa gene expression does not follow the VH paradigm." J Exp Med 169.5 (May 1, 1989): 1859-1864.
Website
http://hdl.handle.net/10161/11491
PMID
2497227
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
169
Issue
5
Publish Date
1989
Start Page
1859
End Page
1864

Mapping of antibody specificities to VH gene families.

VH gene segments represent the products of the repeated duplication and subsequent diversification of a primordial V gene element. It is widely assumed that natural selection, operating via pathogens, has played the dominant role in this process. Here, we screen some 3.7 x 10(4) C mu+ colonies of mitogen-activated B cells for the production of antibodies specific for phosphorylcholine or hen egg lysozyme and expression of the VH X-24, S107, Q52, or J558 gene families. These gene families were expressed at frequencies proportional to their genomic complexity among both unselected and antigen-specific C mu+ colonies. Thus, the capacity to encode equivalent antibody-combining sites is dispersed uniformly among VH families. This result suggests that individual VH genes have not evolved to address specific antigens.

Authors
Kelsoe, G; Miceli, R; Cerny, J; Schulze, DH
MLA Citation
Kelsoe, G, Miceli, R, Cerny, J, and Schulze, DH. "Mapping of antibody specificities to VH gene families." Immunogenetics 29.5 (1989): 288-296.
PMID
2785504
Source
pubmed
Published In
Immunogenetics
Volume
29
Issue
5
Publish Date
1989
Start Page
288
End Page
296

Genotypic analysis of B cell colonies by in situ hybridization. Stoichiometric expression of three VH families in adult C57BL/6 and BALB/c mice.

The filter paper disc method for cloning inducible lymphocytes was used to census the splenic B cell population of C57BL/6 and BALB/c mice for the expression of three VH gene-families, VH X-24, -Q52, and -J558. B cell colonies, arising from single founder lymphocytes, were identified by in situ hybridization with VH family- and C mu-specific cDNA probes. Some 6.7 X 10(4) C mu+ colonies were screened. Among C57BL/6- or BALB/c-derived colonies, approximately 3% were VH X-24+, approximately 19% were VH Q52+, and approximately 54% were VH J558+. These frequencies are consistent with a process of equiprobable expression for individual VH segments, and provide direct evidence that normal splenic B lymphocytes use a process of random genetic combinatorics to generate the antibody repertoire.

Authors
Schulze, DH; Kelsoe, G
MLA Citation
Schulze, DH, and Kelsoe, G. "Genotypic analysis of B cell colonies by in situ hybridization. Stoichiometric expression of three VH families in adult C57BL/6 and BALB/c mice." J Exp Med 166.1 (July 1, 1987): 163-172.
Website
http://hdl.handle.net/10161/10896
PMID
3110348
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
166
Issue
1
Publish Date
1987
Start Page
163
End Page
172

Cloning of mitogen- and antigen-reactive B lymphocytes on filter paper disks: phenotypic and genotypic analysis of B cell colonies.

Authors
Kelsoe, G
MLA Citation
Kelsoe, G. "Cloning of mitogen- and antigen-reactive B lymphocytes on filter paper disks: phenotypic and genotypic analysis of B cell colonies." Methods Enzymol 150 (1987): 287-304.
PMID
3501519
Source
pubmed
Published In
Methods in Enzymology
Volume
150
Publish Date
1987
Start Page
287
End Page
304

Cloning of mitogen- and antigen-reactive B lymphocytes on filter paper discs. II. Paratope frequencies within the mitogen-selected repertoire.

Paratopic frequencies of C57BL/6 (Igh-Vb) and BALB/c (Igh-Va) mice were compared by determining the frequency of lipopolysaccharide-reactive, splenic B lymphocytes secreting antibody specific for (4-hydroxy-5-iodo-3-nitrophenyl) acetyl (NIP), trinitrophenyl (TNP), phosphorylcholine (PC), NIP/TNP, NIP/PC, and sheep erythrocytes. Despite the known genotypic and phenotypic differences between the two Igh-V loci, no significant differences in paratope frequencies were demonstrated. Similar determinations in C.B-20 mice, Ighb congenics of the BALB/c strain, and in C57BL/10 nude mice indicated that the mitogen-generated paratope frequencies directly reflected the capacity of immunoglobulin variable region elements rather than complex interactive or regulatory controls to generate diversity. We conclude that at least for the paratopic repertoire, the role of the somatic processes for the generation of antibody diversity exceeds the influence of germ-line differences between the Ighb and Igha haplotypes.

Authors
Kelsoe, G; Stout, JT
MLA Citation
Kelsoe, G, and Stout, JT. "Cloning of mitogen- and antigen-reactive B lymphocytes on filter paper discs. II. Paratope frequencies within the mitogen-selected repertoire." Cell Immunol 98.2 (April 1, 1986): 506-516.
PMID
3530507
Source
pubmed
Published In
Cellular Immunology
Volume
98
Issue
2
Publish Date
1986
Start Page
506
End Page
516

Regulation of the immune response. II. Concomitant idiotope-specific enhancement and suppression can result in a phenotypically normal response.

Idiotope-specific immunoenhancement or suppression was induced in C57BL/6 mice by the injection of physiological amounts (100 ng-10 micrograms) of monoclonal anti-idiotope antibody. As previously described, nanogram doses enhanced idiotope expression while a 10-micrograms dose of anti-idiotope antibody induced the activation of a population of Thy 1.2+, Lyt 1-, 2+ suppressors. Both positive and negative regulatory activities were confined to the non-mu, idiotope+ compartment of the plaque-forming cell response. Administration of intermediate doses of anti-idiotope antibody resulted in an immune state indistinguishable from that of naive mice. This apparently normal response was in fact the product of a simultaneous activation of balanced enhancing and suppressive activities. When treated with anti-Lyt 2 or Lyt 1 and complement, spleen cell populations taken from such phenotypically "naive" mice revealed latent idiotope-specific immunoenhancement or suppression, demonstrating the components of a functional regulatory equilibrium.

Authors
Kelsoe, G
MLA Citation
Kelsoe, G. "Regulation of the immune response. II. Concomitant idiotope-specific enhancement and suppression can result in a phenotypically normal response." Cell Immunol 98.1 (March 1986): 145-155.
PMID
2943430
Source
pubmed
Published In
Cellular Immunology
Volume
98
Issue
1
Publish Date
1986
Start Page
145
End Page
155

Cloning of mitogen- and antigen-reactive B lymphocytes on filter paper discs. I. A description of the technique and of methods for the analysis of colonies.

A novel technique for establishing short term clones of antigen- or mitogen-activated splenic B lymphocytes is described. Spleen cells are plated onto the surface of filter paper discs and subsequently stimulated by antigen or mitogen in situ; activated B cells proliferate and differentiate into pure colonies of cells analogous to bacterial colonies growing on agar. These colonies of lymphocytes may be characterized in a series of replica hemolytic-plaque, autoradiographic, or immunoenzyme assays making possible a full characterization of the frequency of secreted idiotopes and paratopes and of the cells that produce them. Colony induction by either antigen or mitogen occurs under identical conditions, thus a rigorous comparison between the mitogen-selected and antigen-selected antibody repertoires may be made.

Authors
Kelsoe, G
MLA Citation
Kelsoe, G. "Cloning of mitogen- and antigen-reactive B lymphocytes on filter paper discs. I. A description of the technique and of methods for the analysis of colonies." J Immunol Methods 76.2 (February 11, 1985): 345-363.
PMID
2579160
Source
pubmed
Published In
Journal of Immunological Methods
Volume
76
Issue
2
Publish Date
1985
Start Page
345
End Page
363

Network interactions, 1983.

Authors
Kelsoe, G
MLA Citation
Kelsoe, G. "Network interactions, 1983." Surv Immunol Res 3.2-3 (1984): 169-171.
PMID
6718847
Source
pubmed
Published In
Survey of Immunologic Research
Volume
3
Issue
2-3
Publish Date
1984
Start Page
169
End Page
171

Priority of the anti-idiotypic response after antigen administration: artefact or intriguing network mechanism?

Immunization triggers at least two speck responses measurable at cellular and humoral levels: the antigen-driven proliferation of idiotype-bearing (idt+) cells; and an autochthonous anti-idt response'. As the expansion of the idt* clone(s) is presumed to provide the activating stimulus for the complementary response, one expects the proliferation of anti-idt clones to follow the idt response at an interval equivalent to the time required for ~pmphocyte activation. However, there have been several reports of an autochthonous anti-idt response preceding the expansion of antigen-driven idt' clone(s). In this brief commentary, we examine these reports and offer some suggestions that may resolve this apparent paradox. © 1984.

Authors
Cerny, J; Kelsoe, G
MLA Citation
Cerny, J, and Kelsoe, G. "Priority of the anti-idiotypic response after antigen administration: artefact or intriguing network mechanism?." Immunology Today 5.3 (1984): 61-63.
Source
scival
Published In
Immunology Today
Volume
5
Issue
3
Publish Date
1984
Start Page
61
End Page
63

Induction of immune responses with anti-idiotypic antibodies: implications for the induction of protective immunity.

Authors
Sacks, DL; Kelsoe, GH; Sachs, DH
MLA Citation
Sacks, DL, Kelsoe, GH, and Sachs, DH. "Induction of immune responses with anti-idiotypic antibodies: implications for the induction of protective immunity." Springer Semin Immunopathol 6.1 (1983): 79-97. (Review)
PMID
6412379
Source
pubmed
Published In
Springer seminars in immunopathology
Volume
6
Issue
1
Publish Date
1983
Start Page
79
End Page
97

Network interactions, 1982: audivi, legi, fugi.

Authors
Kelsoe, G
MLA Citation
Kelsoe, G. "Network interactions, 1982: audivi, legi, fugi." Surv Immunol Res 2.3 (1983): 230-232.
PMID
6201980
Source
pubmed
Published In
Survey of Immunologic Research
Volume
2
Issue
3
Publish Date
1983
Start Page
230
End Page
232

Idiotypic determinants used in the analysis of antibody diversification and as regulatory targets.

Authors
Beyreuther, K; Bovens, J; Brüggemann, M; Dildrop, R; Kelsoe, G; Krawinkel, U; Müller, C; Nishikawa, S; Radbruch, A; Reth, M
MLA Citation
Beyreuther, K, Bovens, J, Brüggemann, M, Dildrop, R, Kelsoe, G, Krawinkel, U, Müller, C, Nishikawa, S, Radbruch, A, and Reth, M. "Idiotypic determinants used in the analysis of antibody diversification and as regulatory targets." Ann N Y Acad Sci 418 (1983): 121-129.
PMID
6201100
Source
pubmed
Published In
Annals of the New York Academy of Sciences
Volume
418
Publish Date
1983
Start Page
121
End Page
129

Control of idiotope expression by monoclonal anti-idiotope and idiotope-bearing antibody.

Preinjection of C57BL/6 mice with nano-to microgram amounts of a monoclonal IgG1 antibody directed against a binding site-related idiotope of the anti-NP [(4-hydroxy-3-nitro-phenyl)acetyl] antibody B1-8 results in enhancement or suppression of the corresponding and of another B1-8 idiotope in a subsequent anti-NP response, depending on the dose of the injected anti-idiotope antibody. The enhancing and suppressive effects appear two weeks after anti-idiotope administration and are maximal after 6-8 weeks. They are predominantly expressed at the level of IgG, not IgM, antibodies. Enhancement of idiotype expression, i.e. idiotypic memory, can also be induced by the injection of idiotypic antibody of the IgM class, namely antibody B1-8. This effect might represent one of the general mechanisms by which immunological memory is established.

Authors
Kelsoe, G; Reth, M; Rajewsky, K
MLA Citation
Kelsoe, G, Reth, M, and Rajewsky, K. "Control of idiotope expression by monoclonal anti-idiotope and idiotope-bearing antibody." Eur J Immunol 11.5 (May 1981): 418-423.
PMID
6790289
Source
pubmed
Published In
European Journal of Immunology
Volume
11
Issue
5
Publish Date
1981
Start Page
418
End Page
423
DOI
10.1002/eji.1830110513

Idiotypic regulation by isologous monoclonal anti-idiotope antibodies.

Authors
Reth, M; Kelsoe, G; Rajewsky, K
MLA Citation
Reth, M, Kelsoe, G, and Rajewsky, K. "Idiotypic regulation by isologous monoclonal anti-idiotope antibodies." Nature 290.5803 (March 19, 1981): 257-259.
PMID
6782489
Source
pubmed
Published In
Nature
Volume
290
Issue
5803
Publish Date
1981
Start Page
257
End Page
259

Thymic requirement for cyclical idiotypic and reciprocal anti-idiotypic immune responses to a T-independent antigen.

The role of the thymus in the cyclical appearance of the dominant idiotype of the myeloma protein secreted by the TEPC-15 plasmacytoma (T-15)-bearing plaque-forming cells (PFC) and anti-idiotypic cells (i.e., cells with receptors for T-15) in the spleen during a primary response to the phosphorylcholine determinant of Streptococcus pneumoniae, strain R36a (Pn) was studied using normal mice, thymus-deficient nude mice, and thymus gland-grafted nude mice (TG-nude). The nude mice and their phenotypically normal littermates (LM) were backcrossed on the BALB/c genetic background. The kinetics of the anti-Pn PFC response of BALB/c inbred mice, littermates of nude mice, and TG-nude mice were essentially the same. There was an initial peak on day 5-6 followed by a decline to near background, and then a second peak on day 12. In nude mice, the first peak of anti-Pn PFC (day 5) was comparable in magnitude to that of mice with an intact thymus; however, there was no second peak. In contrast to the cellular response measured at the level of PFC, the serum antibody response to Pn (assayed by passive hemagglutination of sheep erythrocytes coated with Pn polysaccharide) was comparable in all groups of mice and did not show a measurable oscillation. The anti-idiotypic cellular activity was determined by the ability of spleen cells to bind radiolabeled (125I) TEPC-15 myeloma protein (IgA, kappa) which carries an idiotypic determinant indistinguishable from that of most anti-phosphorylcholine antibodies in BALB/c mice. Binding of radiolabeled McPC-603 (IgA, kappa) and MOPC-315 (IgA, lambda 2) myeloma proteins (which lack the T-15 idiotypic determinant) served as controls. The changes in T-15 binding by splenic lymphocytes following the Pn immunization differed between normal and athymic mice. BALB/c, LM, and TG-nude mice showed a biphasic pattern with peaks at days 3--4 and 10--11 that was nearly reciprocal to the PFC curve. On the other hand, T-15 binding in nude mice either declined and remained depressed or was not affected by the ongoing anti-Pn response. These observations demonstrate that mature T cells are required for cyclical idiotypic and anti-idiotypic responses to immunization with a T-independent antigen and suggest that the cyclical immune response may result from an interaction between idiotypic and anti-idiotypic cell clones.

Authors
Kelsoe, G; Isaak, D; Cerny, J
MLA Citation
Kelsoe, G, Isaak, D, and Cerny, J. "Thymic requirement for cyclical idiotypic and reciprocal anti-idiotypic immune responses to a T-independent antigen." J Exp Med 151.2 (February 1, 1980): 289-300.
Website
http://hdl.handle.net/10161/11494
PMID
6965397
Source
pubmed
Published In
The Journal of Experimental Medicine
Volume
151
Issue
2
Publish Date
1980
Start Page
289
End Page
300

Control idiotope expression by monoclonal anti-idiotope antibodies.

Authors
Kelsoe, G; Reth, M; Rajewsky, K
MLA Citation
Kelsoe, G, Reth, M, and Rajewsky, K. "Control idiotope expression by monoclonal anti-idiotope antibodies." Immunol Rev 52 (1980): 75-88. (Review)
PMID
6790422
Source
pubmed
Published In
Immunological Reviews
Volume
52
Publish Date
1980
Start Page
75
End Page
88

Reciprocal expansions of idiotypic and anti-idiotypic clones following antigen stimulation.

Authors
Kelsoe, G; Cerny, J
MLA Citation
Kelsoe, G, and Cerny, J. "Reciprocal expansions of idiotypic and anti-idiotypic clones following antigen stimulation." Nature 279.5711 (May 24, 1979): 333-334.
PMID
313014
Source
pubmed
Published In
Nature
Volume
279
Issue
5711
Publish Date
1979
Start Page
333
End Page
334

A microsporidan contaminant of a nonhuman primate cell culture: ultrastructural comparison with Nosema connori.

Authors
Shadduck, JA; Kelsoe, G; Helmke, RJ
MLA Citation
Shadduck, JA, Kelsoe, G, and Helmke, RJ. "A microsporidan contaminant of a nonhuman primate cell culture: ultrastructural comparison with Nosema connori." J Parasitol 65.1 (February 1979): 185-188.
PMID
109606
Source
pubmed
Published In
Journal of Parasitology
Volume
65
Issue
1
Publish Date
1979
Start Page
185
End Page
188

Immunodiagnosis of infection with Schistosoma mansoni: enzyme-linked immunosorbent assay for detection of antibody to circulating antigen.

A circulating antigen, a negatively charged polysaccharide from the trematode Schistosoma mansoni, was noncovalently bound to the surface of poly(L-lysine)-coated wells in polystyrene trays, which were then used in a micro-enzyme-linked immunosorbent assay (ELISA) test. The method provides an immunodiagnostic test for schistosomiasis of exceptional sensitivity with a high degree of specificity. Comparison of Bell egg counts and ELISA titers revealed a good correlation (r congruent to 0.80) in young individuals with low to moderate worm burdens, but this relationship was less marked in older individuals or those with high egg counts.

Authors
Kelsoe, GH; Weller, TH
MLA Citation
Kelsoe, GH, and Weller, TH. "Immunodiagnosis of infection with Schistosoma mansoni: enzyme-linked immunosorbent assay for detection of antibody to circulating antigen." Proc Natl Acad Sci U S A 75.11 (November 1978): 5715-5717.
PMID
364488
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
75
Issue
11
Publish Date
1978
Start Page
5715
End Page
5717

Regulation of the immune response. I. Regulatory cell equilibrium in the "virgin" state.

Spleen cells from pairs of inbred mice (age- and sex-matched) were challenged in vitro with a T-independent antigen. The specific plaque-forming cell (PFC) response in cultures containing cells from both donors was not intermediate to the responses of independently cultured donor spleen cells. Instead, a pattern of significant suppression (67% of pairs tested) or enhancement (22% of pairs tested) was observed. The suppression or enhancement was antigen-specific and did not represent a general phenomenon within the mixed cultures. A suppressive outcome in mixed cell cultures seems to be associated with cells from the donor with the higher individual PFC response. The converse is true for enhancement. The nonadditive nature of the PFC response in mixed cultures and the frequency of suppression to enhancement (3:1) imply that (a) specific, ongoing immunoregulation occurs even in the naive individual, and (b) this regulation is characterized by regular cycles of specific suppression and enhancement.

Authors
Kelsoe, G; Cerny, J
MLA Citation
Kelsoe, G, and Cerny, J. "Regulation of the immune response. I. Regulatory cell equilibrium in the "virgin" state." Eur J Immunol 8.3 (March 1978): 176-180.
PMID
77784
Source
pubmed
Published In
European Journal of Immunology
Volume
8
Issue
3
Publish Date
1978
Start Page
176
End Page
180
DOI
10.1002/eji.1830080307

The fine structure of spermatogenesis in Hymenolepis diminuta (Cestoda) with a description of the mature spermatozoon.

The processes of spermatogenesis and spermiogenesis in Hymenolepis diminuta were studied by electron microscopy using improved preparative techniques. Spermatogonia (Type A) are characterized by nuclei 3.79 (+/- 0.17) micrometer in diameter, dense cytoplasm packed with free ribosomes and aggregates of mitochondria. After mitoses, certain spermatogonia (Type B) assume syncytial rosettes containing eight nuclei. Primary spermatocytes maintain the rosette syncytium and have large nuclei (4.28 +/- 0.24 micrometer in diameter), smooth endoplasmic reticulum, and polysomes. The secondary spermatocyte is short-lived and is characterized by nuclei (2.0 +/- 0.11 micrometer in diai (2.0 +/- 0.11 micrometer in diameter) and perinuclear membranous lamellae. The syncytial spermatid cluster contains avoid nuclei which condense and elongate to a final diameter of 0.22 +/- 0.04 micrometer. Once elongated, these nuclei become delimited from the syncytium by invaginations of the plasma membrane. During delimitation, cortical peripheral microtubules arise beneath the spermatozoon plasmalemma and a 9 + 1 axoneme extends the length of the mature lance-shaped spermatozoon.

Authors
Kelsoe, GH; Ubelaker, JE; Allison, VF
MLA Citation
Kelsoe, GH, Ubelaker, JE, and Allison, VF. "The fine structure of spermatogenesis in Hymenolepis diminuta (Cestoda) with a description of the mature spermatozoon." Z Parasitenkd 54.2 (December 27, 1977): 175-187.
PMID
605649
Source
pubmed
Published In
Zeitschrift f�r Parasitenkunde
Volume
54
Issue
2
Publish Date
1977
Start Page
175
End Page
187
Show More

Research Areas:

  • Adult
  • Alleles
  • Allergy and Immunology
  • Amino Acids
  • Animals, Genetically Modified
  • Antibodies
  • Antibodies, Monoclonal
  • Antibodies, Viral
  • Antibody Affinity
  • Antibody Diversity
  • Antibody Formation
  • Antibody Specificity
  • Antibody-Producing Cells
  • Antigen-Antibody Complex
  • Antigenic Variation
  • Antigens
  • Antigens, Bacterial
  • Antigens, Surface
  • Antigens, T-Independent
  • Antigens, Viral
  • Apoptosis
  • Arthritis, Rheumatoid
  • Autoantibodies
  • Autoantigens
  • Autoimmune Diseases
  • Autoimmunity
  • B cells
  • B-Lymphocyte Subsets
  • B-Lymphocytes
  • Bacterial Vaccines
  • Binding Sites, Antibody
  • Biotechnology
  • Bone Marrow
  • Bone Marrow Cells
  • CD40 Ligand
  • Cloning, Molecular
  • Complement Activation
  • Complementarity Determining Regions
  • Computational Biology
  • Computer Simulation
  • Crystallography, X-Ray
  • Culture Techniques
  • Cytidine Deaminase
  • Cytokines
  • DNA Repair
  • Dendritic Cells
  • Dendritic Cells, Follicular
  • Dysgammaglobulinemia
  • Enzyme-Linked Immunosorbent Assay
  • Epitope Mapping
  • Epitopes
  • Epitopes, B-Lymphocyte
  • Evolution, Molecular
  • Fetus
  • Flow Cytometry
  • Fluorescent Antibody Technique, Indirect
  • Gene Expression
  • Gene Expression Regulation
  • Gene Expression Regulation, Developmental
  • Gene Rearrangement, B-Lymphocyte
  • Gene Rearrangement, B-Lymphocyte, Heavy Chain
  • Genes, Immunoglobulin
  • Genetic Variation
  • Genotype
  • Germinal Center
  • Granulocytes
  • HIV Antibodies
  • HIV Antigens
  • HIV Envelope Protein gp120
  • HIV Infections
  • HIV-1
  • Haptens
  • Hemagglutinin Glycoproteins, Influenza Virus
  • Hematopoietic Stem Cells
  • Hemolytic Plaque Technique
  • Homeostasis
  • Hyper-IgM Immunodeficiency Syndrome
  • Immune Tolerance
  • Immunity
  • Immunity, Innate
  • Immunization
  • Immunization Schedule
  • Immunogenetics
  • Immunoglobulin Heavy Chains
  • Immunoglobulin Idiotypes
  • Immunoglobulin Isotypes
  • Immunoglobulin Joining Region
  • Immunoglobulin Light Chains
  • Immunoglobulin M
  • Immunoglobulin Switch Region
  • Immunoglobulin Variable Region
  • Immunoglobulin kappa-Chains
  • Immunoglobulin lambda-Chains
  • Immunohistochemistry
  • Immunologic Deficiency Syndromes
  • Immunologic Memory
  • Immunophenotyping
  • Immunosuppression
  • Inflammation
  • Influenza Vaccines
  • Influenza, Human
  • Lipopolysaccharides
  • Lupus Erythematosus, Systemic
  • Lymph Nodes
  • Lymphocyte Activation
  • Lymphocyte Cooperation
  • Lymphoid Tissue
  • Lymphopenia
  • Mice
  • Mice, Inbred Strains
  • Mice, Knockout
  • Mice, Mutant Strains
  • Mice, Transgenic
  • Mitogens
  • Models, Molecular
  • Multigene Family
  • Multipotent Stem Cells
  • Mutation
  • Neutrophils
  • Peyer's Patches
  • Phylogeny
  • Plasma Cells
  • Plasmids
  • Point Mutation
  • Polymerase Chain Reaction
  • Probability
  • Protein Structure, Tertiary
  • Receptors, Antigen, B-Cell
  • Receptors, Complement
  • Receptors, Complement 3b
  • Receptors, Complement 3d
  • Recombinant Proteins
  • Recombination, Genetic
  • Selection, Genetic
  • Sequence Alignment
  • Sequence Homology
  • Somatic Hypermutation, Immunoglobulin
  • Spleen
  • Stochastic Processes
  • Streptococcus pneumoniae
  • Stromal Cells
  • Surface Plasmon Resonance
  • T cells
  • T-Lymphocytes
  • Vaccines
  • Vaccines, Subunit
  • Viremia
  • env Gene Products, Human Immunodeficiency Virus
  • gamma-Globulins