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Markert, Mary Louise

Overview:

Dr. Markert is currently investigating thymus transplantation in complete DiGeorge syndrome. Complete DiGeorge syndrome is a fatal genetic disorder in which patients have heart defects, severe parathyroid hypoplasia and absence of the thymus. In a research protocol complete DiGeorge patients who have no T cells are transplanted with postnatal cultured human thymic epithelial tissue. The transplants are later biopsied to evaluate whether host stem cells have migrated to the tranplanted tissue and developed into T cells. Seventy nine infants with complete DiGeorge anomaly have been transplanted and 56 survive (71%). Her research to date has shown that the patient can develop new recipient T cells in the graft and normal T cell proliferative responses to mitogens and antigens. Thus, in infants born with no T cells because of DiGeorge syndrome, cultured donor postnatal thymic tissue is sufficient for the development of recipient T cells from recipient stem cells. Dr. Markert is now studying patients previously transplanted to learn how long the thymus functions and why the T cell numbers in her post thymus transplantation patients remain low for age - similar to the T cell numbers in patients with partial DiGeorge anomaly who do not need thymus transplantation. In 2012, Dr. Markert began studies in an animal model to use thymus transplantatin to induce tolerance to solid organ transplants. This work was funded by The Hartwell Foundation and is continuing under internal funding.

Positions:

Professor of Pediatrics

Pediatrics, Allergy and Immunology
School of Medicine

Professor of Immunology

Immunology
School of Medicine

Member of the Duke Cancer Institute

Duke Cancer Institute
School of Medicine

Education:

Ph.D. 1981

Ph.D. — Duke University

M.D. 1982

M.D. — Duke University

Grants:

Research Training in Allergy and Clinical Immunology

Administered By
Pediatrics, Allergy and Immunology
AwardedBy
National Institutes of Health
Role
Mentor
Start Date
September 01, 2000
End Date
August 31, 2021

Thymus Transplantation

Administered By
Pediatrics, Allergy and Immunology
AwardedBy
Orphavant Sciences GmbH
Role
Principal Investigator
Start Date
January 01, 2017
End Date
December 31, 2019

An integrated and diverse genomic medicine program for undiagnosed diseases

Administered By
Pediatrics, Medical Genetics
AwardedBy
National Institutes of Health
Role
Collaborator
Start Date
July 01, 2014
End Date
March 31, 2018

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

Thymic Transplantation in Complete DiGeorge Syndrome

Administered By
Pediatrics, Allergy and Immunology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
September 10, 2004
End Date
August 31, 2016

Phase I Serum-free cultured thymus transplantation in DiGeorge anomaly, IND9836, Feb 5, 2008

Administered By
Pediatrics, Allergy and Immunology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
February 15, 2009
End Date
January 31, 2011

Thymic Transplantation in Complete DiGeorge Syndrome

Administered By
Pediatrics, Allergy and Immunology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
September 12, 2009
End Date
August 31, 2010

Dynamics of TCR Repertoire Following Thymus Transplant

Administered By
Pediatrics, Allergy and Immunology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
May 15, 2004
End Date
April 30, 2009

Institutional Training Grant in Pediatric Cardiology

Administered By
Pediatrics, Cardiology
AwardedBy
National Institutes of Health
Role
Mentor
Start Date
April 01, 2004
End Date
March 31, 2009

Dose Study of Thymus Transplantation, IND 9836

Administered By
Pediatrics, Allergy and Immunology
AwardedBy
Food and Drug Administration
Role
Principal Investigator
Start Date
September 30, 2005
End Date
September 29, 2008

Parathyroid and Thymus Transplants in DiGeorge Syndrome

Administered By
Pediatrics, Allergy and Immunology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
July 01, 2004
End Date
June 30, 2006

Telomerase RNA Transfected Dendritic Cell Vaccines

Administered By
Surgery, Urology
AwardedBy
National Institutes of Health
Role
Consultant
Start Date
July 10, 2002
End Date
June 30, 2006

Thymus Transplantation in Complete DiGeorge Syndrome

Administered By
Pediatrics, Allergy and Immunology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
September 24, 1999
End Date
August 31, 2004

Thymic Transplantation in Complete DiGeorge Syndrome

Administered By
Pediatrics, Allergy and Immunology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
September 24, 1999
End Date
June 30, 2004

General Clinical Research Center

Administered By
School of Medicine
AwardedBy
National Institutes of Health
Role
Program Director
Start Date
October 01, 1975
End Date
November 30, 2003

Viral Control and Immune Reconstitution in HIV Infection

Administered By
Surgery, Surgical Sciences
AwardedBy
National Institutes of Health
Role
Co Investigator
Start Date
September 30, 1995
End Date
August 31, 2000

Viral Control And Immune Reconstitution In Hiv Infection

Administered By
Surgery, Surgical Sciences
AwardedBy
National Institutes of Health
Role
Co-Principal Investigator
Start Date
September 01, 1996
End Date
August 31, 1999

Viral Control & Immune Reconstitution In Hiv Infection

Administered By
Surgery
AwardedBy
National Institutes of Health
Role
Co-Principal Investigator
Start Date
September 30, 1995
End Date
August 31, 1999

Mechanisms And Therapy Of Primary Immunodeficiency

Administered By
Pediatrics, Allergy and Immunology
AwardedBy
National Institutes of Health
Role
Co-Principal Investigator
Start Date
August 01, 1996
End Date
July 31, 1999

Mechanisms And Therapy Of Primary Immunodeficiency

Administered By
Pediatrics, Allergy and Immunology
AwardedBy
National Institutes of Health
Role
Co-Principal Investigator
Start Date
August 01, 1995
End Date
July 31, 1999

mechanisms and therapy of primary immunodefficiency

Administered By
Pediatrics, Allergy and Immunology
AwardedBy
National Institutes of Health
Role
Co-Principal Investigator
Start Date
August 01, 1995
End Date
July 31, 1999

Research Training In Allergy And Clinical Immunology

Administered By
Pediatrics, Allergy and Immunology
AwardedBy
National Institutes of Health
Role
Co-Principal Investigator
Start Date
July 19, 1907
End Date
June 30, 1999

General Clinical Research Center (Cru)

Administered By
Medicine, Rheumatology and Immunology
AwardedBy
National Institutes of Health
Role
Co-Principal Investigator
Start Date
October 01, 1975
End Date
November 30, 1998

Center For Developmental Immunology And Host Defense

Administered By
Pediatrics
AwardedBy
National Institutes of Health
Role
Co-Principal Investigator
Start Date
September 01, 1996
End Date
August 31, 1998

Center For Developmental Immunology And Host Defense

Administered By
Pediatrics, Allergy and Immunology
AwardedBy
National Institutes of Health
Role
Co-Principal Investigator
Start Date
April 01, 1992
End Date
August 31, 1997

General Clinical Research Center

Administered By
Medicine, Rheumatology and Immunology
AwardedBy
National Institutes of Health
Role
Co-Principal Investigator
Start Date
December 01, 1993
End Date
November 30, 1995

The Thymus In Immune Reconstitution For Hiv Infection

Administered By
Surgery
AwardedBy
National Institutes of Health
Role
Co-Principal Investigator
Start Date
July 01, 1994
End Date
June 30, 1995

Ada Regulation And The Immune Response: Animal Models

Administered By
Pediatrics, Allergy and Immunology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
April 01, 1994
End Date
March 31, 1995

Ada Regulation And The Immune Response: Animal Models

Administered By
Pediatrics, Allergy and Immunology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
April 01, 1992
End Date
March 31, 1995

Immunodeficiency And Ada Deficiency -- Molecular Analysis

Administered By
Pediatrics, Allergy and Immunology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
July 01, 1990
End Date
June 30, 1991

Immunodeficiency And Ada Deficiency - Molecular Analysis

Administered By
Pediatrics, Allergy and Immunology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
July 01, 1988
End Date
June 01, 1991

Immunodeficiency And Ada Deficiency: Molecular Analysis

Administered By
Pediatrics, Allergy and Immunology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
July 01, 1986
End Date
June 01, 1989
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Publications:

Disseminated Mycobacterium kansasii disease in complete DiGeorge syndrome.

Complete DiGeorge syndrome (cDGS) describes a subset of patients with DiGeorge syndrome that have thymic aplasia, and thus are at risk for severe opportunistic infections. Patients with cDGS and mycobacterial infection have not previously been described. We present this case to illustrate that patients with cDGS are at risk for nontuberculous mycobacterial infections and to discuss further antimicrobial prophylaxis prior to thymic transplantation.A 13-month old male was identified as T cell deficient by the T cell receptor excision circle (TREC) assay on newborn screening, and was subsequently confirmed to have cDGS. He presented with fever and cough, and was treated for chronic aspiration pneumonia as well as Pneumocystis jirovecii infection without significant improvement. It was only after biopsy of mediastinal lymph nodes seen on CT that the diagnosis of disseminated Mycobacterium kansasii was made. We reviewed the literature regarding atypical mycobacterial infections and prophylaxis used in other immunocompromised patients, as well as the current data regarding cDGS detection through TREC newborn screening.Multiple cases of cDGS have been diagnosed via TREC newborn screening, however this is the first patient with cDGS and disseminated mycobacterial infection to be reported in literature. Thymic transplantation is the definitive treatment of choice for cDGS. Prophylaxis with either clarithromycin or azithromycin has been shown to reduce mycobacterial infections in children with advanced human immunodeficiency virus infection.Children with cDGS should receive thymic transplantion as soon as possible, but prior to this are at risk for nontuberculous mycobacterial infections. Severe, opportunistic infections may require invasive testing for diagnosis in patients with cDGS. Antimicrobial prophylaxis should be considered to prevent disseminated mycobacterial infection in these patients.

Authors
Yin, SM; Ferdman, RM; Wang, L; Markert, ML; Tam, JS
MLA Citation
Yin, SM, Ferdman, RM, Wang, L, Markert, ML, and Tam, JS. "Disseminated Mycobacterium kansasii disease in complete DiGeorge syndrome." Journal of clinical immunology 35.5 (July 2015): 435-438.
PMID
26048260
Source
epmc
Published In
Journal of Clinical Immunology
Volume
35
Issue
5
Publish Date
2015
Start Page
435
End Page
438
DOI
10.1007/s10875-015-0171-3

Disseminated Atypical Mycobacterial Infection in Three Patients with Complete Digeorge Anomaly

Authors
Agada, NO; Yin, SM; Tam, JS; Kelly, MS; Markert, ML
MLA Citation
Agada, NO, Yin, SM, Tam, JS, Kelly, MS, and Markert, ML. "Disseminated Atypical Mycobacterial Infection in Three Patients with Complete Digeorge Anomaly." February 2015.
Source
wos-lite
Published In
Journal of Allergy and Clinical Immunology
Volume
135
Issue
2
Publish Date
2015
Start Page
AB10
End Page
AB10

Complete thymectomy in adult rats with non-invasive endotracheal intubation.

Thymectomy in neonatal rodents is an established and reliable procedure for immunological studies. However, in adult rats, complications of hemorrhage and pneumothorax from pleural disruption can result in a significant mortality rate. This protocol is a simple method of rat thymectomy that utilizes a mini-sternotomy and endotracheal intubation. Intubation is accomplished with a non-invasive and easily reproducible method and allows for positive pressure ventilation to prevent pneumothorax and a controlled airway that allows sufficient time for careful thymus dissection to minimize pleural disruption. A 1.5 cm sternal incision decreases contact with mediastinal vessels and pleura, while still providing full visualization of the thymus. Following exposure of the mediastinum, the thymus is removed by blunt dissection under magnification. The pleural space is then sealed by suture closure of the pre-tracheal muscles followed by the application of surgical glue. The thorax is then closed by suture closure of the sternum, followed by suture closure of the skin. All thymectomies were complete as evidenced by immunohistochemical (IHC) staining of mediastinal tissue, and absence of naïve T-cells by flow cytometry, and the procedure had a 96% survival rate. This method is suitable when complete thymectomy with minimal complications is desired for further immunological studies in athymic adult rats.

Authors
Rendell, VR; Giamberardino, C; Li, J; Markert, ML; Brennan, TV
MLA Citation
Rendell, VR, Giamberardino, C, Li, J, Markert, ML, and Brennan, TV. "Complete thymectomy in adult rats with non-invasive endotracheal intubation." Journal of visualized experiments : JoVE 94 (December 29, 2014).
PMID
25590868
Source
epmc
Published In
Journal of Visualized Experiments
Issue
94
Publish Date
2014
DOI
10.3791/52152

Clinical course and outcome predictors of critically ill infants with complete DiGeorge anomaly following thymus transplantation.

To identify risk factors for PICU admission and mortality of infants with complete DiGeorge anomaly treated with thymus transplantation. We hypothesized that age at transplantation and the presence of congenital heart disease would be risk factors for emergent PICU admission, and these factors plus development of septicemia would increase morbidity and mortality.Retrospective review.Academic medical-surgical PICU.All infants with complete DiGeorge anomaly treated with thymus transplantation between January 1, 1993, and July 1, 2010.None.Consent was obtained from 71 infants with complete DiGeorge anomaly for thymus transplantation, and 59 infants were transplanted. Median age at transplantation was 5.0 months (range, 1.1-22.1 mo). After transplantation, 12 of 59 infants (20%) required 25 emergent PICU admissions. Seven of 12 infants (58%) survived to PICU discharge with six surviving 6 months posttransplantation. Forty-two of 59 infants (71%) transplanted had congenital heart disease, and 9 of 12 (75%) who were admitted to the PICU had congenital heart disease. In 15 of 25 admissions (60%), intubation and mechanical ventilation were necessary. There was no difference between median ventilation-free days between infants with and without congenital heart disease (33 d vs 23 d, p = 0.544). There was also no correlation between ventilation-free days and age of transplantation (R, 0.17; p = 0.423). Age at transplantation and the presence of congenital heart disease were not associated with risk for PICU admission (odds ratio, 0.95; 95% CI, 0.78-1.15 and odds ratio, 1.27; 95% CI, 0.30-5.49, respectively) or PICU mortality (odds ratio, 0.98; 95% CI, 0.73-1.31 and odds ratio, 0.40; 95% CI, 0.15-1.07, respectively).Most transplanted infants did not require emergent PICU admission. Age at transplantation and the presence of congenital heart disease were not associated with PICU admission or mortality.

Authors
Lee, JH; Markert, ML; Hornik, CP; McCarthy, EA; Gupton, SE; Cheifetz, IM; Turner, DA
MLA Citation
Lee, JH, Markert, ML, Hornik, CP, McCarthy, EA, Gupton, SE, Cheifetz, IM, and Turner, DA. "Clinical course and outcome predictors of critically ill infants with complete DiGeorge anomaly following thymus transplantation." Pediatric critical care medicine : a journal of the Society of Critical Care Medicine and the World Federation of Pediatric Intensive and Critical Care Societies 15.7 (September 2014): e321-e326.
PMID
25068252
Source
epmc
Published In
Pediatric Critical Care Medicine
Volume
15
Issue
7
Publish Date
2014
Start Page
e321
End Page
e326
DOI
10.1097/pcc.0000000000000219

Defects in Thymic Development: DiGeorge/CHARGE/Chromosome 22q11.2 Deletion

© 2014 Elsevier Inc. All rights reserved.DiGeorge anomaly is characterized by defects of the heart, thymus, and parathyroid. To make the diagnosis of DiGeorge anomaly, two of the three organs must be affected. The genetic and syndromic associations with DiGeorge anomaly include 22q11.2 deletion syndrome, CHARGE (coloboma, heart defects, atresia of the choanae, retardation of growth and development, genital hypoplasia, ear anomalies and/or deafness) syndrome, and diabetic embryopathy. Most infants with DiGeorge anomaly have a small thymus resulting in low T cell numbers. Less than 1% of infants with DiGeorge anomaly are athymic and consequently have no thymically-derived T cells. This condition is "complete" DiGeorge anomaly. These infants are at high risk for life-threatening infections. Some infants with complete DiGeorge anomaly may develop an atypical phenotype similar to Omenn syndrome, with circulating oligoclonal T cells and severe rash. The symptoms leading to the diagnosis of DiGeorge anomaly and the management of these patients are described in this chapter.

Authors
Markert, ML
MLA Citation
Markert, ML. "Defects in Thymic Development: DiGeorge/CHARGE/Chromosome 22q11.2 Deletion." Stiehm's Immune Deficiencies. August 12, 2014. 221-242.
Source
scopus
Publish Date
2014
Start Page
221
End Page
242
DOI
10.1016/B978-0-12-405546-9.00008-X

Thymus Transplantation

© 2014 Elsevier Inc. All rights reserved.Thymus transplantation is an experimental procedure in which allogeneic, cultured, postnatal thymus tissue is transplanted into an athymic infant. Almost all transplant recipients are infants with complete DiGeorge anomaly. The donor thymus is obtained as discarded tissue from pediatric heart surgery after informed consent. It is sectioned, and maintained in culture while donor screening is performed. After 2-3 weeks in culture, the thymus tissue is transplanted into the quadriceps muscle of the athymic infant under general anesthesia in the operating room. Temporary immunosuppression is required in approximately half of transplant recipients. In a series of 67 consecutive patients, the 1-year survival rate was 76%. The median age of survivors is 7.8 years. Naïve CD4+ and CD8+ T cells develop over the first 6-9 months. Immunosuppression, immunoglobulin replacement therapy, and Pneumocystis prophylaxis are discontinued at approximately 1 year post-transplantation.

Authors
Louise Markert, M
MLA Citation
Louise Markert, M. "Thymus Transplantation." (August 12, 2014): 1059-1067. (Chapter)
Source
scopus
Publish Date
2014
Start Page
1059
End Page
1067
DOI
10.1016/B978-0-12-405546-9.00060-1

Human syndromes of immunodeficiency and dysregulation are characterized by distinct defects in T-cell receptor repertoire development.

BACKGROUND: Human immunodeficiencies characterized by hypomorphic mutations in critical developmental and signaling pathway genes allow for the dissection of the role of these genes in the development of the T-cell receptor (TCR) repertoire and the correlation of alterations of the TCR repertoire with diverse clinical phenotypes. OBJECTIVE: The presence of T cells in patients with Omenn syndrome (OS) and patients with atypical presentations of severe combined immunodeficiency gene mutations presents an opportunity to study the effects of the causal genes on TCR repertoires and provides a window into the clinical heterogeneity observed. METHODS: We performed deep sequencing of TCRβ complementarity-determining region 3 (CDR3) regions in subjects with a series of immune dysregulatory conditions caused by mutations in recombination activating gene 1/2 (RAG 1/2), IL-2 receptor γ (IL2RG), and ζ chain-associated protein kinase 70 (ZAP70); a patient with atypical DiGeorge syndrome; and healthy control subjects. RESULTS: We found that patients with OS had marked reductions in TCRβ diversity compared with control subjects, as expected. Patients with atypical presentations of RAG or IL2RG mutations associated with autoimmunity and granulomatous disease did not have altered overall diversity but instead had skewed V-J pairing and skewed CDR3 amino acid use. Although germline TCRs were more abundant and clonally expanded in patients with OS, nongermline sequences were expanded as well. TCRβ from patients with RAG mutations had less junctional diversity and smaller CDR3s than patients with OS caused by other gene mutations and healthy control subjects but relatively similar CDR3 amino acid use. CONCLUSIONS: High-throughput TCR sequencing of rare immune disorders has demonstrated that quantitative TCR diversity can appear normal despite qualitative changes in repertoire and strongly suggests that in human subjects RAG enzymatic function might be necessary for normal CDR3 junctional diversity.

Authors
Yu, X; Almeida, JR; Darko, S; van der Burg, M; DeRavin, SS; Malech, H; Gennery, A; Chinn, I; Markert, ML; Douek, DC; Milner, JD
MLA Citation
Yu, X, Almeida, JR, Darko, S, van der Burg, M, DeRavin, SS, Malech, H, Gennery, A, Chinn, I, Markert, ML, Douek, DC, and Milner, JD. "Human syndromes of immunodeficiency and dysregulation are characterized by distinct defects in T-cell receptor repertoire development." J Allergy Clin Immunol 133.4 (April 2014): 1109-1115.
PMID
24406074
Source
pubmed
Published In
Journal of Allergy and Clinical Immunology
Volume
133
Issue
4
Publish Date
2014
Start Page
1109
End Page
1115
DOI
10.1016/j.jaci.2013.11.018

Human Syndromes of Immune Deficiency and Dysregulation are Characterized by Distinct Defects in T-Cell Receptor Repertoire Development

Authors
Yu, X; Almeida, J; Darko, S; van der Burg, M; DeRavin, SS; Malech, H; Gennery, A; Chinn, I; Markert, ML; Perales, M-A; Pamer, EG; Douek, DC; Milner, JD
MLA Citation
Yu, X, Almeida, J, Darko, S, van der Burg, M, DeRavin, SS, Malech, H, Gennery, A, Chinn, I, Markert, ML, Perales, M-A, Pamer, EG, Douek, DC, and Milner, JD. "Human Syndromes of Immune Deficiency and Dysregulation are Characterized by Distinct Defects in T-Cell Receptor Repertoire Development." April 2014.
Source
wos-lite
Published In
Journal of Clinical Immunology
Volume
34
Issue
3
Publish Date
2014
Start Page
350
End Page
351

Human syndromes of immunodeficiency and dysregulation are characterized by distinct defects in T-cell receptor repertoire development

Background Human immunodeficiencies characterized by hypomorphic mutations in critical developmental and signaling pathway genes allow for the dissection of the role of these genes in the development of the T-cell receptor (TCR) repertoire and the correlation of alterations of the TCR repertoire with diverse clinical phenotypes. Objective The presence of T cells in patients with Omenn syndrome (OS) and patients with atypical presentations of severe combined immunodeficiency gene mutations presents an opportunity to study the effects of the causal genes on TCR repertoires and provides a window into the clinical heterogeneity observed. Methods We performed deep sequencing of TCRβ complementarity-determining region 3 (CDR3) regions in subjects with a series of immune dysregulatory conditions caused by mutations in recombination activating gene 1/2 (RAG 1/2), IL-2 receptor γ (IL2RG), and ζ chain-associated protein kinase 70 (ZAP70); a patient with atypical DiGeorge syndrome; and healthy control subjects. Results We found that patients with OS had marked reductions in TCRβ diversity compared with control subjects, as expected. Patients with atypical presentations of RAG or IL2RG mutations associated with autoimmunity and granulomatous disease did not have altered overall diversity but instead had skewed V-J pairing and skewed CDR3 amino acid use. Although germline TCRs were more abundant and clonally expanded in patients with OS, nongermline sequences were expanded as well. TCRβ from patients with RAG mutations had less junctional diversity and smaller CDR3s than patients with OS caused by other gene mutations and healthy control subjects but relatively similar CDR3 amino acid use. Conclusions High-throughput TCR sequencing of rare immune disorders has demonstrated that quantitative TCR diversity can appear normal despite qualitative changes in repertoire and strongly suggests that in human subjects RAG enzymatic function might be necessary for normal CDR3 junctional diversity. © 2013 American Academy of Allergy, Asthma & Immunology.

Authors
Yu, X; Almeida, JR; Darko, S; Van Der Burg, M; Deravin, SS; Malech, H; Gennery, A; Chinn, I; Markert, ML; Douek, DC; Milner, JD
MLA Citation
Yu, X, Almeida, JR, Darko, S, Van Der Burg, M, Deravin, SS, Malech, H, Gennery, A, Chinn, I, Markert, ML, Douek, DC, and Milner, JD. "Human syndromes of immunodeficiency and dysregulation are characterized by distinct defects in T-cell receptor repertoire development." Journal of Allergy and Clinical Immunology 133.4 (January 1, 2014).
Source
scopus
Published In
Journal of Allergy and Clinical Immunology
Volume
133
Issue
4
Publish Date
2014
DOI
10.1016/j.jaci.2013.11.018

Complete thymectomy in adult rats with non-invasive endotracheal intubation

Thymectomy in neonatal rodents is an established and reliable procedure for immunological studies. However, in adult rats, complications of hemorrhage and pneumothorax from pleural disruption can result in a significant mortality rate. This protocol is a simple method of rat thymectomy that utilizes a mini-sternotomy and endotracheal intubation. Intubation is accomplished with a non-invasive and easily reproducible method and allows for positive pressure ventilation to prevent pneumothorax and a controlled airway that allows sufficient time for careful thymus dissection to minimize pleural disruption. A 1.5 cm sternal incision decreases contact with mediastinal vessels and pleura, while still providing full visualization of the thymus. Following exposure of the mediastinum, the thymus is removed by blunt dissection under magnification. The pleural space is then sealed by suture closure of the pre-tracheal muscles followed by the application of surgical glue. The thorax is then closed by suture closure of the sternum, followed by suture closure of the skin. All thymectomies were complete as evidenced by immunohistochemical (IHC) staining of mediastinal tissue, and absence of naïve T-cells by flow cytometry, and the procedure had a 96% survival rate. This method is suitable when complete thymectomy with minimal complications is desired for further immunological studies in athymic adult rats.

Authors
Rendell, VR
MLA Citation
Rendell, VR. "Complete thymectomy in adult rats with non-invasive endotracheal intubation." Journal of visualized experiments : JoVE 94 (January 1, 2014).
Source
scopus
Published In
Journal of Visualized Experiments
Issue
94
Publish Date
2014
DOI
10.3791/52152

Clinical course and outcome predictors of critically Ill infants with complete DiGeorge anomaly following thymus transplantation

Copyright © 2014 by the Society of Critical Care Medicine and the World Federation of Pediatric Intensive and Critical Care Societies.Objectives: To identify risk factors for PICU admission and mortality of infants with complete DiGeorge anomaly treated with thymus transplantation. We hypothesized that age at transplantation and the presence of congenital heart disease would be risk factors for emergent PICU admission, and these factors plus development of septicemia would increase morbidity and mortality. Design: Retrospective review. Setting: Academic medical-surgical PICU. Patients: All infants with complete DiGeorge anomaly treated with thymus transplantation between January 1, 1993, and July 1, 2010. Interventions: None. Measurements and Main Results: Consent was obtained from 71 infants with complete DiGeorge anomaly for thymus transplantation, and 59 infants were transplanted. Median age at transplantation was 5.0 months (range, 1.1-22.1 mo). After transplantation, 12 of 59 infants (20%) required 25 emergent PICU admissions. Seven of 12 infants (58%) survived to PICU discharge with six surviving 6 months posttransplantation. Forty-two of 59 infants (71%) transplanted had congenital heart disease, and 9 of 12 (75%) who were admitted to the PICU had congenital heart disease. In 15 of 25 admissions (60%), intubation and mechanical ventilation were necessary. There was no difference between median ventilation-free days between infants with and without congenital heart disease (33 d vs 23 d, p = 0.544). There was also no correlation between ventilation-free days and age of transplantation (R, 0.17; p = 0.423). Age at transplantation and the presence of congenital heart disease were not associated with risk for PICU admission (odds ratio, 0.95; 95% CI, 0.78-1.15 and odds ratio, 1.27; 95% CI, 0.30-5.49, respectively) or PICU mortality (odds ratio, 0.98; 95% CI, 0.73-1.31 and odds ratio, 0.40; 95% CI, 0.15-1.07, respectively). Conclusions: Most transplanted infants did not require emergent PICU admission. Age at transplantation and the presence of congenital heart disease were not associated with PICU admission or mortality.

Authors
Lee, JH; Markert, ML; Hornik, CP; McCarthy, EA; Gupton, SE; Cheifetz, IM; Turner, DA
MLA Citation
Lee, JH, Markert, ML, Hornik, CP, McCarthy, EA, Gupton, SE, Cheifetz, IM, and Turner, DA. "Clinical course and outcome predictors of critically Ill infants with complete DiGeorge anomaly following thymus transplantation." Pediatric Critical Care Medicine 15.7 (January 1, 2014): e321-e326.
Source
scopus
Published In
Pediatric Critical Care Medicine
Volume
15
Issue
7
Publish Date
2014
Start Page
e321
End Page
e326
DOI
10.1097/PCC.0000000000000219

SEPSIS INCREASES MORBIDITY OF CRITICALLY ILL INFANTS WITH COMPLETE DIGEORGE ANOMALY TREATED WITH THYMUS TRANSPLANTATION

Authors
Lee, JH; Markert, ML; Hornik, CP; McCarthy, EA; Gupton, SE; Cheifetz, IM; Turner, DA
MLA Citation
Lee, JH, Markert, ML, Hornik, CP, McCarthy, EA, Gupton, SE, Cheifetz, IM, and Turner, DA. "SEPSIS INCREASES MORBIDITY OF CRITICALLY ILL INFANTS WITH COMPLETE DIGEORGE ANOMALY TREATED WITH THYMUS TRANSPLANTATION." October 2013.
Source
wos-lite
Published In
Intensive Care Medicine
Volume
39
Publish Date
2013
Start Page
S412
End Page
S412

Thymus transplantation restores the repertoires of forkhead box protein 3 (FoxP3)+ and FoxP3- T cells in complete DiGeorge anomaly.

The development of T cells with a regulatory phenotype after thymus transplantation has not been examined previously in complete DiGeorge anomaly (cDGA). Seven athymic infants with cDGA and non-maternal pretransplantation T cell clones were assessed. Pretransplantation forkhead box protein 3 (Foxp3)(+) T cells were detected in five of the subjects. Two subjects were studied in greater depth. T cell receptor variable β chain (TCR-Vβ) expression was assessed by flow cytometry. In both subjects, pretransplantation FoxP3(+) and total CD4(+) T cells showed restricted TCR-Vβ expression. The development of naive T cells and diverse CD4(+) TCR-Vβ repertoires following thymic transplantation indicated successful thymopoiesis from the thymic tissue grafts. Infants with atypical cDGA develop rashes and autoimmune phenomena before transplantation, requiring treatment with immunosuppression, which was discontinued successfully subsequent to the observed thymopoiesis. Post-transplantation, diverse TCR-Vβ family expression was also observed in FoxP3(+) CD4(+) T cells. Interestingly, the percentages of each of the TCR-Vβ families expressed on FoxP3(+) and total CD4(+) T cells differed significantly between these T lymphocyte subpopulations before transplantation. By 16 months post-transplantation, however, the percentages of expression of each TCR-Vβ family became significantly similar between FoxP3(+) and total CD4(+) T cells. Sequencing of TCRBV DNA confirmed the presence of clonally amplified pretransplantation FoxP3(+) and FoxP3(-) T cells. After thymus transplantation, increased polyclonality was observed for both FoxP3(+) and FoxP3(-) cells, and pretransplantation FoxP3(+) and FoxP3(-) clonotypes essentially disappeared. Thus, post-transplantation thymic function was associated with the development of a diverse repertoire of FoxP3(+) T cells in cDGA, corresponding with immunological and clinical recovery.

Authors
Chinn, IK; Milner, JD; Scheinberg, P; Douek, DC; Markert, ML
MLA Citation
Chinn, IK, Milner, JD, Scheinberg, P, Douek, DC, and Markert, ML. "Thymus transplantation restores the repertoires of forkhead box protein 3 (FoxP3)+ and FoxP3- T cells in complete DiGeorge anomaly." Clin Exp Immunol 173.1 (July 2013): 140-149.
PMID
23607606
Source
pubmed
Published In
Clinical & Experimental Immunology
Volume
173
Issue
1
Publish Date
2013
Start Page
140
End Page
149
DOI
10.1111/cei.12088

Quantification of total T-cell receptor diversity by flow cytometry and spectratyping

Background: T-cell receptor diversity correlates with immune competency and is of particular interest in patients undergoing immune reconstitution. Spectratyping generates data about T-cell receptor CDR3 length distribution for each BV gene but is technically complex. Flow cytometry can also be used to generate data about T-cell receptor BV gene usage, but its utility has not been compared to or tested in combination with spectratyping.Results: Using flow cytometry and spectratype data, we have defined a divergence metric that quantifies the deviation from normal of T-cell receptor repertoire. We have shown that the sample size is a sensitive parameter in the predicted flow divergence values, but not in the spectratype divergence values. We have derived two ways to correct for the measurement bias using mathematical and statistical approaches and have predicted a lower bound in the number of lymphocytes needed when using the divergence as a substitute for diversity.Conclusions: Using both flow cytometry and spectratyping of T-cells, we have defined the divergence measure as an indirect measure of T-cell receptor diversity. We have shown the dependence of the divergence measure on the sample size before it can be used to make predictions regarding the diversity of the T-cell receptor repertoire. © 2013 Ciupe et al.; licensee BioMed Central Ltd.

Authors
Ciupe, SM; Devlin, BH; Markert, ML; Kepler, TB
MLA Citation
Ciupe, SM, Devlin, BH, Markert, ML, and Kepler, TB. "Quantification of total T-cell receptor diversity by flow cytometry and spectratyping." BMC Immunology 14.1 (2013).
PMID
23914737
Source
scival
Published In
BMC Immunology
Volume
14
Issue
1
Publish Date
2013
DOI
10.1186/1471-2172-14-35

Thymus Defects/Transplant and Problems Associated with Immune Reconstitution

Authors
Markert, ML; Devlin, BH; McCarthy, EA
MLA Citation
Markert, ML, Devlin, BH, and McCarthy, EA. "Thymus Defects/Transplant and Problems Associated with Immune Reconstitution." 2013.
Source
wos-lite
Published In
PROCEEDINGS OF THE 15TH MEETING OF THE EUROPEAN SOCIETY IMMUNODEFICIENCIES (ESID)
Publish Date
2013
Start Page
99
End Page
104

Thymic reconstitution

Authors
Markert, ML; Devlin, BH; McCarthy, EA
MLA Citation
Markert, ML, Devlin, BH, and McCarthy, EA. "Thymic reconstitution." Clinical Immunology: Principles and Practice: Fourth Edition. December 12, 2012. 1032-1038.
Source
scopus
Publish Date
2012
Start Page
1032
End Page
1038
DOI
10.1016/B978-0-7234-3691-1.00023-4

Diagnosis of 22q11.2 deletion syndrome and artemis deficiency in two children with T-B-NK+ immunodeficiency.

Two infants are described who presented with 22q11.2 deletion and a T(-)B(-)NK(+) immune phenotype. For both infants, the initial diagnosis was athymia secondary to complete DiGeorge anomaly. The first infant underwent thymus transplantation but 6 months after transplantation had circulating thymus donor T cells; the patient did not develop recipient naïve T cells. Genetic analyses revealed that both patients had Artemis deficiency, a rare form of severe combined immunodeficiency (SCID). Both infants have subsequently undergone bone marrow transplantation. These cases illustrate the importance and paradox of differentiating SCID from complete DiGeorge anomaly because hematopoietic stem cell transplantation (HSCT) is the preferred treatment for SCID but is ineffective for complete DiGeorge anomaly. However, if the thymus is completely absent, donor stem cells from a HSCT would not be able to be educated.

Authors
Heimall, J; Keller, M; Saltzman, R; Bunin, N; McDonald-McGinn, D; Zakai, E; de Villartay, J-P; Moshous, D; Ariue, B; McCarthy, EA; Devlin, BH; Parikh, S; Buckley, RH; Markert, ML
MLA Citation
Heimall, J, Keller, M, Saltzman, R, Bunin, N, McDonald-McGinn, D, Zakai, E, de Villartay, J-P, Moshous, D, Ariue, B, McCarthy, EA, Devlin, BH, Parikh, S, Buckley, RH, and Markert, ML. "Diagnosis of 22q11.2 deletion syndrome and artemis deficiency in two children with T-B-NK+ immunodeficiency." J Clin Immunol 32.5 (October 2012): 1141-1144.
PMID
22864628
Source
pubmed
Published In
Journal of Clinical Immunology
Volume
32
Issue
5
Publish Date
2012
Start Page
1141
End Page
1144
DOI
10.1007/s10875-012-9741-9

THYMUS TRANSPLANTATION FOR COMPLETE DIGEORGE SYNDROME: THE LONDON EXPERIENCE

Authors
Davies, EG; Gilmour, KC; Parsley, K; Curry, J; Sebire, N; Poliani, L; McCarthy, EA; Devlin, B; Markert, ML; Thrasher, AJ
MLA Citation
Davies, EG, Gilmour, KC, Parsley, K, Curry, J, Sebire, N, Poliani, L, McCarthy, EA, Devlin, B, Markert, ML, and Thrasher, AJ. "THYMUS TRANSPLANTATION FOR COMPLETE DIGEORGE SYNDROME: THE LONDON EXPERIENCE." September 2012.
Source
wos-lite
Published In
Journal of Clinical Immunology
Volume
32
Publish Date
2012
Start Page
12
End Page
13

Human FOXN1-deficiency is associated with αβ double-negative and FoxP3+ T-cell expansions that are distinctly modulated upon thymic transplantation.

Forkhead box N1 (FOXN1) is a transcription factor crucial for thymic epithelium development and prevention of its involution. Investigation of a patient with a rare homozygous FOXN1 mutation (R255X), leading to alopecia universalis and thymus aplasia, unexpectedly revealed non-maternal circulating T-cells, and, strikingly, large numbers of aberrant double-negative αβ T-cells (CD4negCD8neg, DN) and regulatory-like T-cells. These data raise the possibility that a thymic rudiment persisted, allowing T-cell development, albeit with disturbances in positive/negative selection, as suggested by DN and FoxP3+ cell expansions. Although regulatory-like T-cell numbers normalized following HLA-mismatched thymic transplantation, the αβDN subset persisted 5 years post-transplantation. Involution of thymus allograft likely occurred 3 years post-transplantation based on sj/βTREC ratio, which estimates intrathymic precursor T-cell divisions and, consequently, thymic explant output. Nevertheless, functional immune-competence was sustained, providing new insights for the design of immunological reconstitution strategies based on thymic transplantation, with potential applications in other clinical settings.

Authors
Albuquerque, AS; Marques, JG; Silva, SL; Ligeiro, D; Devlin, BH; Dutrieux, J; Cheynier, R; Pignata, C; Victorino, RMM; Markert, ML; Sousa, AE
MLA Citation
Albuquerque, AS, Marques, JG, Silva, SL, Ligeiro, D, Devlin, BH, Dutrieux, J, Cheynier, R, Pignata, C, Victorino, RMM, Markert, ML, and Sousa, AE. "Human FOXN1-deficiency is associated with αβ double-negative and FoxP3+ T-cell expansions that are distinctly modulated upon thymic transplantation." PLoS One 7.5 (2012): e37042-.
PMID
22590644
Source
pubmed
Published In
PloS one
Volume
7
Issue
5
Publish Date
2012
Start Page
e37042
DOI
10.1371/journal.pone.0037042

Interpreting low T-cell receptor excision circles in newborns with DiGeorge anomaly: importance of assessing naive T-cell markers.

Authors
Knutsen, AP; Baker, MW; Markert, ML
MLA Citation
Knutsen, AP, Baker, MW, and Markert, ML. "Interpreting low T-cell receptor excision circles in newborns with DiGeorge anomaly: importance of assessing naive T-cell markers." J Allergy Clin Immunol 128.6 (December 2011): 1375-1376. (Letter)
PMID
22018899
Source
pubmed
Published In
Journal of Allergy and Clinical Immunology
Volume
128
Issue
6
Publish Date
2011
Start Page
1375
End Page
1376
DOI
10.1016/j.jaci.2011.08.019

Thymic microenvironment reconstitution after postnatal human thymus transplantation.

A functional thymus develops after cultured thymus tissue is transplanted into subjects with complete DiGeorge anomaly. To gain insight into how the process occurs, 7 post-transplantation thymus biopsy tissues were evaluated. In 5 of 7 biopsies, the thymus appeared to be predominantly cortex with thymocytes expressing cortical markers. Unexpectedly, the epithelium expressed both cortical [cortical dendritic reticulum antigen 2 (CDR2)] and medullary [cytokeratin (CK) 14] markers. Early medullary development was suggested by epithelial cell adhesion molecule (EpCAM) reactivity in small areas of biopsies. Two other biopsies had distinct mature cortex and medulla with normal restriction of CK14 to the medulla and subcapsular cortex, and of CDR2 to cortex. These data are consistent with a model in which thymic epithelium contains CK14+ "progenitor epithelial cells". After transplantation these cells proliferate as CK14+CDR2+ thymic epithelial cells that are associated with cortical thymocytes. Later these cells differentiate into distinct cortical and medullary epithelia.

Authors
Li, B; Li, J; Devlin, BH; Markert, ML
MLA Citation
Li, B, Li, J, Devlin, BH, and Markert, ML. "Thymic microenvironment reconstitution after postnatal human thymus transplantation." Clin Immunol 140.3 (September 2011): 244-259.
PMID
21565561
Source
pubmed
Published In
Clinical Immunology
Volume
140
Issue
3
Publish Date
2011
Start Page
244
End Page
259
DOI
10.1016/j.clim.2011.04.004

Successful extracorporeal membrane oxygenation for respiratory failure in an infant with DiGeorge anomaly, following thymus transplantation.

We report the first successful use of venovenous extracorporeal membrane oxygenation (ECMO) for refractory respiratory failure in an infant with DiGeorge anomaly, following thymus transplantation. A 23-month-old female with complete immune-incompetent DiGeorge anomaly 65 days after allogenic thymus transplantation was treated in our pediatric intensive care unit for acute respiratory failure secondary to bacterial sepsis. She subsequently developed acute hypercarbic respiratory failure unresponsive to conventional medical therapy. She was successfully managed with venovenous ECMO for 4 days, with complete resolution of her respiratory symptoms. This case demonstrates the complex decision making process regarding initiation of ECMO in patients with severe immunodeficiency.

Authors
Hornik, CP; Hartman, ME; Markert, ML; Lodge, AJ; Cheifetz, IM; Turner, DA
MLA Citation
Hornik, CP, Hartman, ME, Markert, ML, Lodge, AJ, Cheifetz, IM, and Turner, DA. "Successful extracorporeal membrane oxygenation for respiratory failure in an infant with DiGeorge anomaly, following thymus transplantation." Respiratory care 56.6 (June 2011): 866-870.
PMID
21333090
Source
epmc
Published In
Respiratory Care
Volume
56
Issue
6
Publish Date
2011
Start Page
866
End Page
870
DOI
10.4187/respcare.01051

Induction of tolerance to parental parathyroid grafts using allogeneic thymus tissue in patients with DiGeorge anomaly.

DiGeorge anomaly can affect both thymic and parathyroid function. Although athymia is corrected by allogeneic thymus transplantation, treatment options for hypoparathyroidism have been unsatisfactory. Parathyroid transplantation offers the potential for definitive cure but remains challenging because of graft rejection. Some allogeneic parathyroid grafts have functioned in adult recipients in the context of immunosuppression for renal transplantation. Other efforts have attempted to reduce the allogenicity of the parathyroid grafts through manipulation of the parathyroid tissues before transplantation (by using encapsulation or special culture techniques). Recently, we demonstrated the efficacy of parental parathyroid transplantation when combined with allogeneic thymus transplantation in an infant with complete DiGeorge anomaly. The recipient developed tolerance toward the parathyroid donor. The parathyroid graft has functioned for 5 years after transplantation without the need for continued immunosuppression or calcium supplementation. We observed that matching of the allogeneic thymus graft to the parathyroid donor HLA class II alleles that are unshared with the recipient appears to be associated with the induction of tolerance toward the parathyroid graft. Further work is needed to determine the optimal means for using combined allogeneic thymus and parental parathyroid transplantation to correct hypoparathyroidism in patients with both complete and partial DiGeorge anomaly.

Authors
Chinn, IK; Markert, ML
MLA Citation
Chinn, IK, and Markert, ML. "Induction of tolerance to parental parathyroid grafts using allogeneic thymus tissue in patients with DiGeorge anomaly." J Allergy Clin Immunol 127.6 (June 2011): 1351-1355. (Review)
PMID
21513969
Source
pubmed
Published In
Journal of Allergy and Clinical Immunology
Volume
127
Issue
6
Publish Date
2011
Start Page
1351
End Page
1355
DOI
10.1016/j.jaci.2011.03.033

First use of thymus transplantation therapy for FOXN1 deficiency (nude/SCID): a report of 2 cases.

FOXN1 deficiency is a primary immunodeficiency characterized by athymia, alopecia totalis, and nail dystrophy. Two infants with FOXN1 deficiency were transplanted with cultured postnatal thymus tissue. Subject 1 presented with disseminated Bacillus Calmette-Guérin infection and oligoclonal T cells with no naive markers. Subject 2 had respiratory failure, human herpes virus 6 infection, cytopenias, and no circulating T cells. The subjects were given thymus transplants at 14 and 9 months of life, respectively. Subject 1 received immunosuppression before and for 10 months after transplantation. With follow up of 4.9 and 2.9 years, subjects 1 and 2 are well without infectious complications. The pretransplantation mycobacterial disease in subject 1 and cytopenias in subject 2 resolved. Subject 2 developed autoimmune thyroid disease 1.6 years after transplantation. Both subjects developed functional immunity. Subjects 1 and 2 have 1053/mm(3) and 1232/mm(3) CD3(+) cells, 647/mm(3) and 868/mm(3) CD4(+) T cells, 213/mm(3) and 425/mm(3) naive CD4(+) T cells, and 10 200 and 5700 T-cell receptor rearrangement excision circles per 100 000 CD3(+) cells, respectively. They have normal CD4 T-cell receptor β variable repertoires. Both subjects developed antigen-specific proliferative responses and have discontinued immunoglobulin replacement. In summary, thymus transplantation led to T-cell reconstitution and function in these FOXN1 deficient infants.

Authors
Markert, ML; Marques, JG; Neven, B; Devlin, BH; McCarthy, EA; Chinn, IK; Albuquerque, AS; Silva, SL; Pignata, C; de Saint Basile, G; Victorino, RM; Picard, C; Debre, M; Mahlaoui, N; Fischer, A; Sousa, AE
MLA Citation
Markert, ML, Marques, JG, Neven, B, Devlin, BH, McCarthy, EA, Chinn, IK, Albuquerque, AS, Silva, SL, Pignata, C, de Saint Basile, G, Victorino, RM, Picard, C, Debre, M, Mahlaoui, N, Fischer, A, and Sousa, AE. "First use of thymus transplantation therapy for FOXN1 deficiency (nude/SCID): a report of 2 cases." Blood 117.2 (January 13, 2011): 688-696.
PMID
20978268
Source
pubmed
Published In
Blood
Volume
117
Issue
2
Publish Date
2011
Start Page
688
End Page
696
DOI
10.1182/blood-2010-06-292490

Mechanisms of tolerance to parental parathyroid tissue when combined with human allogeneic thymus transplantation.

BACKGROUND: The induction of tolerance toward third-party solid organ grafts with allogeneic thymus tissue transplantation has not been previously demonstrated in human subjects. OBJECTIVE: Infants with complete DiGeorge anomaly (having neither thymus nor parathyroid function) were studied for conditions and mechanisms required for the development of tolerance to third-party solid organ tissues. METHODS: Four infants who met the criteria received parental parathyroid with allogeneic thymus transplantation and were studied. RESULTS: Two of 3 survivors showed function of both grafts but subsequently lost parathyroid function. They demonstrated alloreactivity against the parathyroid donor in mixed lymphocyte cultures. For these 2 recipients, parathyroid donor HLA class II alleles were mismatched with the recipient and thymus. MHC class II tetramers confirmed the presence of recipient CD4(+) T cells with specificity toward a mismatched parathyroid donor class II allele. The third survivor has persistent graft function and lacks alloreactivity toward the parathyroid donor. All parathyroid donor class II alleles were shared with either the recipient or the thymus graft, with minor differences between the parathyroid (HLA-DRB1∗1104) and thymus (HLA-DRB1∗1101). Tetramer analyses detected recipient T cells specific for the parathyroid HLA-DRB1∗1104 allele. Alloreactivity toward the parathyroid donor was restored with low doses of IL-2. CONCLUSION: Tolerance toward parathyroid grafts in combined parental parathyroid and allogeneic thymus transplantation requires matching of thymus tissue to parathyroid HLA class II alleles to promote negative selection and suppression of recipient T cells that have alloreactivity toward the parathyroid grafts. This matching strategy may be applied toward tolerance induction in future combined thymus and solid organ transplantation efforts.

Authors
Chinn, IK; Olson, JA; Skinner, MA; McCarthy, EA; Gupton, SE; Chen, D-F; Bonilla, FA; Roberts, RL; Kanariou, MG; Devlin, BH; Markert, ML
MLA Citation
Chinn, IK, Olson, JA, Skinner, MA, McCarthy, EA, Gupton, SE, Chen, D-F, Bonilla, FA, Roberts, RL, Kanariou, MG, Devlin, BH, and Markert, ML. "Mechanisms of tolerance to parental parathyroid tissue when combined with human allogeneic thymus transplantation." J Allergy Clin Immunol 126.4 (October 2010): 814-820.e8.
PMID
20832849
Source
pubmed
Published In
Journal of Allergy and Clinical Immunology
Volume
126
Issue
4
Publish Date
2010
Start Page
814
End Page
820.e8
DOI
10.1016/j.jaci.2010.07.016

Thymus transplantation.

Thymus transplantation is a promising investigational therapy for infants born with no thymus. Because of the athymia, these infants lack T cell development and have a severe primary immunodeficiency. Although thymic hypoplasia or aplasia is characteristic of DiGeorge anomaly, in "complete" DiGeorge anomaly, there is no detectable thymus as determined by the absence of naive (CD45RA(+), CD62L(+)) T cells. Transplantation of postnatal allogeneic cultured thymus tissue was performed in sixty subjects with complete DiGeorge anomaly who were under the age of 2 years. Recipient survival was over 70%. Naive T cells developed 3-5 months after transplantation. The graft recipients were able to discontinue antibiotic prophylaxis, and immunoglobulin replacement. Immunosuppression was used in a subset of subjects but was discontinued when naive T cells developed. The adverse events have been acceptable with thyroid disease being the most common. Research continues on mechanisms underlying immune reconstitution after thymus transplantation.

Authors
Markert, ML; Devlin, BH; McCarthy, EA
MLA Citation
Markert, ML, Devlin, BH, and McCarthy, EA. "Thymus transplantation." Clin Immunol 135.2 (May 2010): 236-246. (Review)
PMID
20236866
Source
pubmed
Published In
Clinical Immunology
Volume
135
Issue
2
Publish Date
2010
Start Page
236
End Page
246
DOI
10.1016/j.clim.2010.02.007

Mechanisms of Tolerance to Parental Parathyroid Tissue when Combined with Human Allogeneic Thymus Transplantation

Authors
Chinn, IK; Olson, JA; Skinner, MA; McCarthy, EA; Gupton, SE; Bonilla, FA; Roberts, RL; Kanariou, MG; Devlin, BH; Markert, ML
MLA Citation
Chinn, IK, Olson, JA, Skinner, MA, McCarthy, EA, Gupton, SE, Bonilla, FA, Roberts, RL, Kanariou, MG, Devlin, BH, and Markert, ML. "Mechanisms of Tolerance to Parental Parathyroid Tissue when Combined with Human Allogeneic Thymus Transplantation." February 2010.
Source
wos-lite
Published In
Journal of Allergy and Clinical Immunology
Volume
125
Issue
2
Publish Date
2010
Start Page
AB55
End Page
AB55

Comèl-Netherton syndrome defined as primary immunodeficiency.

BACKGROUND: Mutations in serine protease inhibitor Kazal-type 5 (SPINK5), encoding the serine protease inhibitor lympho-epithelial Kazal-type 5 related inhibitor (LEKTI), cause Comèl-Netherton syndrome, an autosomal-recessive disease characterized by congenital ichthyosis, bamboo hair, and atopic diathesis. Despite increased frequency of infections, the immunocompetence of patients with Comèl-Netherton syndrome has not been extensively investigated. OBJECTIVE: To define Comèl-Netherton syndrome as a primary immunodeficiency disorder and to explore the benefit of intravenous immunoglobulin replacement therapy. METHODS: We enrolled 9 patients with Comèl-Netherton syndrome, sequenced SPINK5, and analyzed LEKTI expression by immunohistochemistry. Immune function was assessed by measuring cognate immunity, serum cytokine levels, and natural killer cell cytotoxicity. RESULTS: All patients presented with recurrent skin infections caused predominantly by Staphylococcus aureus. All but 1 reported recurrent respiratory tract infections; 78% had sepsis and/or pneumonia; 67% had recurrent gastrointestinal disease and failure to thrive. Mutations in SPINK5-including 6 novel mutations-were identified in 8 patients. LEKTI expression was decreased or absent in all patients. Immunologic evaluation revealed reduced memory B cells and defective responses to vaccination with Pneumovax and bacteriophage phiX174, characterized by impaired antibody amplification and class-switching. Immune dysregulation was suggested by a skewed T(h)1 phenotype and elevated proinflammatory cytokine levels, whereas serum concentrations of the chemokine (C-C motif) ligand 5 and natural killer cell cytotoxicity were decreased. Treatment with intravenous immunoglobulin resulted in remarkable clinical improvement and temporarily increased natural killer cell cytotoxicity. CONCLUSION: These data provide new insights into the immunopathology of Comèl-Netherton syndrome and demonstrate that this multisystem disorder, characterized by lack of LEKTI expression in epithelial cells, is complicated by cognate and innate immunodeficiency that responds favorably to intravenous immunoglobulin therapy.

Authors
Renner, ED; Hartl, D; Rylaarsdam, S; Young, ML; Monaco-Shawver, L; Kleiner, G; Markert, ML; Stiehm, ER; Belohradsky, BH; Upton, MP; Torgerson, TR; Orange, JS; Ochs, HD
MLA Citation
Renner, ED, Hartl, D, Rylaarsdam, S, Young, ML, Monaco-Shawver, L, Kleiner, G, Markert, ML, Stiehm, ER, Belohradsky, BH, Upton, MP, Torgerson, TR, Orange, JS, and Ochs, HD. "Comèl-Netherton syndrome defined as primary immunodeficiency." J Allergy Clin Immunol 124.3 (September 2009): 536-543.
PMID
19683336
Source
pubmed
Published In
Journal of Allergy and Clinical Immunology
Volume
124
Issue
3
Publish Date
2009
Start Page
536
End Page
543
DOI
10.1016/j.jaci.2009.06.009

The dynamics of T-cell receptor repertoire diversity following thymus transplantation for DiGeorge anomaly.

T cell populations are regulated both by signals specific to the T-cell receptor (TCR) and by signals and resources, such as cytokines and space, that act independently of TCR specificity. Although it has been demonstrated that disruption of either of these pathways has a profound effect on T-cell development, we do not yet have an understanding of the dynamical interactions of these pathways in their joint shaping of the T cell repertoire. Complete DiGeorge Anomaly is a developmental abnormality that results in the failure of the thymus to develop, absence of T cells, and profound immune deficiency. After receiving thymic tissue grafts, patients suffering from DiGeorge anomaly develop T cells derived from their own precursors but matured in the donor tissue. We followed three DiGeorge patients after thymus transplantation to utilize the remarkable opportunity these subjects provide to elucidate human T-cell developmental regulation. Our goal is the determination of the respective roles of TCR-specific vs. TCR-nonspecific regulatory signals in the growth of these emerging T-cell populations. During the course of the study, we measured peripheral blood T-cell concentrations, TCRbeta V gene-segment usage and CDR3-length spectratypes over two years or more for each of the subjects. We find, through statistical analysis based on a novel stochastic population-dynamic T-cell model, that the carrying capacity corresponding to TCR-specific resources is approximately 1000-fold larger than that of TCR-nonspecific resources, implying that the size of the peripheral T-cell pool at steady state is determined almost entirely by TCR-nonspecific mechanisms. Nevertheless, the diversity of the TCR repertoire depends crucially on TCR-specific regulation. The estimated strength of this TCR-specific regulation is sufficient to ensure rapid establishment of TCR repertoire diversity in the early phase of T cell population growth, and to maintain TCR repertoire diversity in the face of substantial clonal expansion-induced perturbation from the steady state.

Authors
Ciupe, SM; Devlin, BH; Markert, ML; Kepler, TB
MLA Citation
Ciupe, SM, Devlin, BH, Markert, ML, and Kepler, TB. "The dynamics of T-cell receptor repertoire diversity following thymus transplantation for DiGeorge anomaly." PLoS Comput Biol 5.6 (June 2009): e1000396-.
PMID
19521511
Source
pubmed
Published In
PLoS computational biology
Volume
5
Issue
6
Publish Date
2009
Start Page
e1000396
DOI
10.1371/journal.pcbi.1000396

Thymus transplantation in complete DiGeorge anomaly.

Complete DiGeorge anomaly is characterized by athymia, congenital heart disease, and hypoparathyroidism. This congenital disease is fatal by age 2 years unless immune reconstitution is successful. There are multiple underlying syndromes associated with complete DiGeorge anomaly including 22q11 hemizygosity in approximately 50%, CHARGE association in approximately 25%, and diabetic embryopathy in approximately 15%. Approximately one-third of patients present with rash and lymphadenopathy associated with oligoclonal "host" T cells. This condition resembles Omenn syndrome. Immunosuppression is necessary to control the oligoclonal T cells. The results of thymus transplantation are reported for a series of 50 patients, of whom 36 survive. The survivors develop naïve T cells and a diverse T cell repertoire.

Authors
Markert, ML; Devlin, BH; Chinn, IK; McCarthy, EA
MLA Citation
Markert, ML, Devlin, BH, Chinn, IK, and McCarthy, EA. "Thymus transplantation in complete DiGeorge anomaly." Immunol Res 44.1-3 (2009): 61-70.
PMID
19066739
Source
pubmed
Published In
Immunologic Research
Volume
44
Issue
1-3
Publish Date
2009
Start Page
61
End Page
70
DOI
10.1007/s12026-008-8082-5

Characterization of cultured thymus tissue used for transplantation with emphasis on promiscuous expression of thyroid tissue-specific genes.

Autoimmune thyroid disease occurs in some complete DiGeorge anomaly patients after thymus transplantation. This study was designed to assess the effect of culture of thymus tissue on the expression of genes involved in the development of autoimmunity. The expression of autoimmune regulator (AIRE), thyroglobulin (TG), thyroid peroxidase (TPO), and cytokeratin RNAs was examined in thymocytes and thymus tissue on the day of thymus harvest and after 14 and 21 days of culture. Immunohistochemistry was used to evaluate the cytokeratin expression in the thymus tissue. AIRE, TG, TPO, and cytokeratin mRNAs were found in harvest-day, 14-day and 21-day cultured tissues. Levels of AIRE, TG, and cytokeratin mRNAs were mostly higher after culture compared to expression on the harvest day, likely secondary to thymocyte depletion.

Authors
Li, B; Li, J; Hsieh, C-S; Hale, LP; Li, Y-J; Devlin, BH; Markert, ML
MLA Citation
Li, B, Li, J, Hsieh, C-S, Hale, LP, Li, Y-J, Devlin, BH, and Markert, ML. "Characterization of cultured thymus tissue used for transplantation with emphasis on promiscuous expression of thyroid tissue-specific genes." Immunol Res 44.1-3 (2009): 71-83.
PMID
19066738
Source
pubmed
Published In
Immunologic Research
Volume
44
Issue
1-3
Publish Date
2009
Start Page
71
End Page
83
DOI
10.1007/s12026-008-8083-4

Thymus transplantation

Thymus transplantation was first attempted in the 1960s and 1970s using fetal thymus tissue [1, 2]. The results overall were disappointing [3-6]. In part the poor outcomes related to the lack of reagents needed to characterize and identify the patients into those who were truly athymic (complete DiGeorge anomaly) and those who had bone marrow stem cell problems (severe combined immunodeficiency). It is also possible that the fetal thymus tissue was too small to reconstitute a human infant [7]. The use of fetal thymus carried the risk of fatal graft versus host disease since mature T-cells can be found in the human thymus by the end of the first trimester [3]. By 1986, in a review of 26 infants treated with fetal thymus transplantation, 22 had died; the other 4 patients had achieved a 3-year survival [6]. © 2008 Springer Milan.

Authors
Markert, ML; Devlin, BH; McCarthy, EA; Chinn, IK; Hale, LP
MLA Citation
Markert, ML, Devlin, BH, McCarthy, EA, Chinn, IK, and Hale, LP. "Thymus transplantation." (December 1, 2008): 255-267. (Chapter)
Source
scopus
Publish Date
2008
Start Page
255
End Page
267
DOI
10.1007/978-88-470-0828-1_30

Factors affecting success of thymus transplantation for complete DiGeorge anomaly.

Thymus transplantation shows promise for the treatment of athymia in complete DiGeorge anomaly. This report reviews the effects of dose of thymus tissue, ABO compatibility, HLA matching, culture conditions, age of donor and immunosuppression of recipient on immune outcomes at 1 year after transplantation. Forty-nine athymic subjects have been treated with cultured postnatal allogeneic thymus tissue; 36 (73%) survive with only one subject on immunosuppression at 1.5 years. Of 31 surviving subjects more than 1 year after transplantation, 30 (97%) developed naive T cells, T-cell proliferative responses to mitogens and a diverse T-cell receptor beta variable (TCRBV) repertoire. The dose of thymus tissue, HLA matching and use of immunosuppression had nonsignificant effects on these outcome variables. Removal of deoxyguanosine from culture medium and length of culture did not adversely affect outcomes. Use of thymus tissue from donors over 1 month of age, versus under 1 month, resulted in higher total T-cell numbers (p = 0.03). However, this finding must be confirmed in a prospective trial. Although subtle immune effects may yet be associated with some of the factors tested, it is remarkable that consistently good immune outcomes result despite variation in dose, HLA matching and use of immunosuppression.

Authors
Markert, ML; Devlin, BH; Chinn, IK; McCarthy, EA; Li, YJ
MLA Citation
Markert, ML, Devlin, BH, Chinn, IK, McCarthy, EA, and Li, YJ. "Factors affecting success of thymus transplantation for complete DiGeorge anomaly." Am J Transplant 8.8 (August 2008): 1729-1736.
PMID
18557726
Source
pubmed
Published In
American Journal of Transplantation
Volume
8
Issue
8
Publish Date
2008
Start Page
1729
End Page
1736
DOI
10.1111/j.1600-6143.2008.02301.x

Use of allograft biopsies to assess thymopoiesis after thymus transplantation.

Thymus allograft biopsies were performed in athymic infants with complete DiGeorge anomaly after thymus transplantation to assess whether the thymus allograft tissue was able to support thymopoiesis. Forty-four consecutive infants were treated with postnatal cultured thymus allografts. Thirty biopsies and six autopsies evaluating the allograft site were obtained in 33 infants, 23 of whom survive. The allograft was examined by immunohistochemistry for evidence of thymopoiesis. Grafted thymus tissue was found in 25 of 30 biopsies, 23 of which showed thymopoiesis. All 19 survivors with thymopoiesis on biopsy developed naive T cells and T cell function. Autopsies were done in six subjects, three of whom had biopsies. All autopsy samples contained thymus tissue including one for which the biopsy had not contained graft. Of the six autopsies, one had evidence of thymopoiesis. Epithelium without thymopoiesis was seen in two of 25 biopsies in which thymus tissue was detected and in five of six autopsies. Graft rejection was seen in one autopsy. Biopsies were important for showing the following: 1) the damaging effect of pulse steroids on thymopoiesis; 2) the need for adequate immunosuppression of atypical subjects; and 3) the presence of thymopoiesis in the presence of ongoing immunosuppression. In addition, the biopsy could rule out graft rejection in the atypical subjects who had oligoclonal T cells that could cause rejection. In summary, combining biopsy and autopsy data, allogeneic thymus tissues showed thymopoiesis in 24 of 29 (86%) evaluable transplants. The results of these biopsies led to improved care of these complex patients.

Authors
Markert, ML; Li, J; Devlin, BH; Hoehner, JC; Rice, HE; Skinner, MA; Li, Y-J; Hale, LP
MLA Citation
Markert, ML, Li, J, Devlin, BH, Hoehner, JC, Rice, HE, Skinner, MA, Li, Y-J, and Hale, LP. "Use of allograft biopsies to assess thymopoiesis after thymus transplantation." J Immunol 180.9 (May 1, 2008): 6354-6364.
PMID
18424759
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
180
Issue
9
Publish Date
2008
Start Page
6354
End Page
6364

Treatment of infants with complete DiGeorge anomaly.

Authors
Markert, ML
MLA Citation
Markert, ML. "Treatment of infants with complete DiGeorge anomaly." J Allergy Clin Immunol 121.4 (April 2008): 1063-. (Letter)
PMID
18289653
Source
pubmed
Published In
Journal of Allergy and Clinical Immunology
Volume
121
Issue
4
Publish Date
2008
Start Page
1063
DOI
10.1016/j.jaci.2007.12.1181

The cutaneous manifestations of atypical complete DiGeorge syndrome: a histopathologic and immunohistochemical study.

DiGeorge syndrome is a congenital anomaly with a constellation of findings that includes thymic hypoplasia. Only a small subset of patients with DiGeorge syndrome has complete athymia, classified as complete DiGeorge anomaly; one third of these patients show an eczematous dermatitis, oligoclonal T-cells and lymphadenopathy, known as atypical complete DiGeorge anomaly. Six biopsies from six patients with the distinctive clinical phenotype of atypical complete DiGeorge anomaly were studied. Every biopsy showed exocytosis (100%), parakeratosis, often confluent and spongiosis (100%). Neutrophilic abscesses (50%), dyskeratosis (67%) and satellite cell necrosis (50%) were seen. Perieccrine and perivascular inflammation were seen in half of the cases. Eosinophils were identified (83%); most commonly in both the epidermis and dermis. All of lymphocytes were CD3 positive. Most (83%) of cases contained T-cell intracellular antigen 1 (TIA-1) positive cells. Special testing of the selected patients using spectratyping identified oligoclonal T-cell populations. The presence of dyskeratotic keratinocytes, satellite cell necrosis and parakeratotic scale with neutrophils characterizes the cutaneous rash seen in this subset of complete DiGeorge syndrome patients. Such skin lesions from patients with DiGeorge anomaly should alert the pathologist to the potential diagnosis of atypical complete DiGeorge anomaly. The pathophysiologic role of the oligoclonal T-cells in this entity requires additional study.

Authors
Selim, MA; Markert, ML; Burchette, JL; Herman, CM; Turner, JW
MLA Citation
Selim, MA, Markert, ML, Burchette, JL, Herman, CM, and Turner, JW. "The cutaneous manifestations of atypical complete DiGeorge syndrome: a histopathologic and immunohistochemical study." J Cutan Pathol 35.4 (April 2008): 380-385.
PMID
18333898
Source
pubmed
Published In
Journal of Cutaneous Pathology
Volume
35
Issue
4
Publish Date
2008
Start Page
380
End Page
385
DOI
10.1111/j.1600-0560.2007.00816.x

Long-term tolerance to allogeneic thymus transplants in complete DiGeorge anomaly.

Thymus transplantation in subjects with complete DiGeorge anomaly using postnatal allogeneic HLA-nonmatched cultured thymus tissue provides immunoreconstitution. Tolerance of the newly developed T cells toward the donor thymus has not previously been studied. Mixed lymphocyte cultures were used to test 12 thymus transplant recipients for long-term tolerance toward their thymus allografts. Two subjects tested for responses toward thymus donor peripheral blood mononuclear cells showed significantly less reactivity toward the donors compared to responses against third-party allogeneic cells. Peripheral blood mononuclear cells from 10 other subjects were less responsive toward cryopreserved donor thymic cells than toward allogeneic cells (P=0.00007). Adult control peripheral blood mononuclear cells proliferated strongly in response to the donor thymic cells. Both the subjects and controls showed similar proliferative responses against allogeneic cells and phytohemagglutinin. This study provides in vitro evidence for long-term tolerance of complete DiGeorge anomaly thymus transplantation recipients toward their HLA-nonmatched thymus grafts.

Authors
Chinn, IK; Devlin, BH; Li, Y-J; Markert, ML
MLA Citation
Chinn, IK, Devlin, BH, Li, Y-J, and Markert, ML. "Long-term tolerance to allogeneic thymus transplants in complete DiGeorge anomaly." Clin Immunol 126.3 (March 2008): 277-281.
PMID
18155964
Source
pubmed
Published In
Clinical Immunology
Volume
126
Issue
3
Publish Date
2008
Start Page
277
End Page
281
DOI
10.1016/j.clim.2007.11.009

Combined thymus and parathyroid allotransplantation in complete DiGeorge anomaly

Authors
Chinni, IK; Jr, OJA; Skinner, MA; McCarthy, EA; Gupton, SE; Bonilla, FA; Roberts, RL; Kanariou, MG; Devlin, BH; Markert, ML
MLA Citation
Chinni, IK, Jr, OJA, Skinner, MA, McCarthy, EA, Gupton, SE, Bonilla, FA, Roberts, RL, Kanariou, MG, Devlin, BH, and Markert, ML. "Combined thymus and parathyroid allotransplantation in complete DiGeorge anomaly." February 2008.
Source
wos-lite
Published In
Journal of Allergy and Clinical Immunology
Volume
121
Issue
2
Publish Date
2008
Start Page
S269
End Page
S269
DOI
10.1016/j.jaci.2007.12.1067

Thymic reconstitution

Authors
Markert, ML; Devlin, BH
MLA Citation
Markert, ML, and Devlin, BH. "Thymic reconstitution." Clinical Immunology (2008): 1253-1261.
Source
scival
Published In
Clinical Immunology
Publish Date
2008
Start Page
1253
End Page
1261
DOI
10.1016/B978-0-323-04404-2.10084-3

Human T cell reconstitution in DiGeorge syndrome and HIV-1 infection.

The thymus is essential for proper development and maintenance of a broad T cell repertoire capable of recognizing a wide-range of foreign antigens. Recent advances in multicolor flow cytometry, non-invasive imaging techniques, and molecular assessments of thymic function have enabled a more comprehensive characterization of human thymic output in clinical settings than in the past. These techniques have been particularly valuable in monitoring human T cells after therapeutic thymic grafting for complete DiGeorge syndrome and during HIV-1 infection and AIDS. By defining the degree and mechanisms of T cell reconstitution in these settings, clinical investigators and primary caregivers have been able to better diagnose, treat and care for individuals with congenital or acquired immune deficiencies associated with loss of thymic function.

Authors
Hudson, LL; Louise Markert, M; Devlin, BH; Haynes, BF; Sempowski, GD
MLA Citation
Hudson, LL, Louise Markert, M, Devlin, BH, Haynes, BF, and Sempowski, GD. "Human T cell reconstitution in DiGeorge syndrome and HIV-1 infection." Semin Immunol 19.5 (October 2007): 297-309. (Review)
PMID
18035553
Source
pubmed
Published In
Seminars in Immunology
Volume
19
Issue
5
Publish Date
2007
Start Page
297
End Page
309
DOI
10.1016/j.smim.2007.10.002

Review of 54 patients with complete DiGeorge anomaly enrolled in protocols for thymus transplantation: outcome of 44 consecutive transplants.

The purpose of this study was to characterize a large group of infants with complete DiGeorge anomaly and to evaluate the ability of thymus transplantation to reconstitute immune function in these infants. DiGeorge anomaly is characterized by varying defects of the heart, thymus, and parathyroid glands. Complete DiGeorge anomaly refers to the subgroup that is athymic (< 1%). The characteristics of 54 subjects at presentation and results from 44 consecutive thymus transplantations are reported. Remarkably, only 52% had 22q11 hemizygosity and only 57% had congenital heart disease requiring surgery. Thirty-one percent developed an atypical phenotype with rash and lymphadenopathy. To date, 33 of 44 subjects who received a transplant survive (75%) with post-transplantation follow-up as long as 13 years. All deaths occurred within 12 months of transplantation. All 25 subjects who were tested 1 year after transplantation had developed polyclonal T-cell repertoires and proliferative responses to mitogens. Adverse events developing after transplantation included hypothyroidism in 5 subjects and enteritis in 1 subject. In summary, diagnosis of complete DiGeorge anomaly is challenging because of the variability of presentation. Thymus transplantation was well tolerated and resulted in stable immunoreconstitution in these infants.

Authors
Markert, ML; Devlin, BH; Alexieff, MJ; Li, J; McCarthy, EA; Gupton, SE; Chinn, IK; Hale, LP; Kepler, TB; He, M; Sarzotti, M; Skinner, MA; Rice, HE; Hoehner, JC
MLA Citation
Markert, ML, Devlin, BH, Alexieff, MJ, Li, J, McCarthy, EA, Gupton, SE, Chinn, IK, Hale, LP, Kepler, TB, He, M, Sarzotti, M, Skinner, MA, Rice, HE, and Hoehner, JC. "Review of 54 patients with complete DiGeorge anomaly enrolled in protocols for thymus transplantation: outcome of 44 consecutive transplants." Blood 109.10 (May 15, 2007): 4539-4547.
PMID
17284531
Source
pubmed
Published In
Blood
Volume
109
Issue
10
Publish Date
2007
Start Page
4539
End Page
4547
DOI
10.1182/blood-2006-10-048652

Tolerance to allogeneic thymus transplantation in complete DiGeorge syndrome

Authors
Chinn, IK; Devlin, BH; Alexieff, MJ; Li, J; Gupton, SE; McCarthy, EA; Markert, ML
MLA Citation
Chinn, IK, Devlin, BH, Alexieff, MJ, Li, J, Gupton, SE, McCarthy, EA, and Markert, ML. "Tolerance to allogeneic thymus transplantation in complete DiGeorge syndrome." January 2007.
Source
wos-lite
Published In
Journal of Allergy and Clinical Immunology
Volume
119
Issue
1
Publish Date
2007
Start Page
S12
End Page
S12
DOI
10.1016/j.jaci.2006.11.062

Development of regulatory T cells after thymus transplantation in subjects with complete DiGeorge syndrome

Authors
Chinn, I; Devlin, B; Markert, ML
MLA Citation
Chinn, I, Devlin, B, and Markert, ML. "Development of regulatory T cells after thymus transplantation in subjects with complete DiGeorge syndrome." 2007.
Source
wos-lite
Published In
Clinical Immunology
Volume
123
Publish Date
2007
Start Page
S42
End Page
S43
DOI
10.1016/j.clim.2007.03.297

New developments in primary immunodeficiencies: Report on the 2006 CIS Primary Immunodeficiency Diseases Consortium Conference, June 1, 2006.

Authors
Cunningham Rundles, C; Fleisher, T; Markert, ML; Orange, J; Ochs, H; Sullivan, K
MLA Citation
Cunningham Rundles, C, Fleisher, T, Markert, ML, Orange, J, Ochs, H, and Sullivan, K. "New developments in primary immunodeficiencies: Report on the 2006 CIS Primary Immunodeficiency Diseases Consortium Conference, June 1, 2006." Clin Immunol 121.3 (December 2006): 369-371. (Letter)
PMID
16861041
Source
pubmed
Published In
Clinical Immunology
Volume
121
Issue
3
Publish Date
2006
Start Page
369
End Page
371
DOI
10.1016/j.clim.2006.06.004

Parathyroid gland transplantation without posttransplantation immunosuppression

Authors
Chinn, IK; Devlin, BH; Alexieff, MA; Li, J; McCarthy, E; Skinner, MA; Olson, JA; Markert, ML
MLA Citation
Chinn, IK, Devlin, BH, Alexieff, MA, Li, J, McCarthy, E, Skinner, MA, Olson, JA, and Markert, ML. "Parathyroid gland transplantation without posttransplantation immunosuppression." February 2006.
Source
wos-lite
Published In
Journal of Allergy and Clinical Immunology
Volume
117
Issue
2
Publish Date
2006
Start Page
S108
End Page
S108
DOI
10.1016/j.jaci.2005.12.431

SpA: web-accessible spectratype analysis: data management, statistical analysis and visualization.

SUMMARY: SpA is a web-accessible system for the management, visualization and statistical analysis of T-cell receptor spectratype data. Users upload data from their spectratype analyzers to SpA, which saves the raw data and user-defined supplementary covariates to a secure database. The statistical engine performs several data analyses and statistical summaries. The visualization engine displays spectratype histograms in a Java applet and in an image file suitable for download. All of these results are also saved to the database and remain accessible to the user. Additional statistical tools specific to the analysis of multiple spectratypes are also available through the SpA interface. AVAILABILITY: The service is freely accessible via the web at http://www.duke.edu/~kepler/spa.html. Additional technical support and specialized statistical analysis and consultation are available by arrangement with the authors and, depending on the service requested, may be subject to fee.

Authors
He, M; Tomfohr, JK; Devlin, BH; Sarzotti, M; Markert, ML; Kepler, TB
MLA Citation
He, M, Tomfohr, JK, Devlin, BH, Sarzotti, M, Markert, ML, and Kepler, TB. "SpA: web-accessible spectratype analysis: data management, statistical analysis and visualization." Bioinformatics 21.18 (September 15, 2005): 3697-3699.
PMID
16051675
Source
pubmed
Published In
Bioinformatics
Volume
21
Issue
18
Publish Date
2005
Start Page
3697
End Page
3699
DOI
10.1093/bioinformatics/bti600

Parathyroid and thymus transplantation in complete DiGeorge syndrome.

Authors
Markert, ML; Devlin, BH; Alexieff, MA; Li, J; McCarthy, E; Skinner, MA; Olson, JA
MLA Citation
Markert, ML, Devlin, BH, Alexieff, MA, Li, J, McCarthy, E, Skinner, MA, and Olson, JA. "Parathyroid and thymus transplantation in complete DiGeorge syndrome." September 2005.
Source
wos-lite
Published In
Clinical Immunology
Volume
116
Issue
3
Publish Date
2005
Start Page
294
End Page
295

Time course of normalization of T cell receptor repertoire in a complete DiGeorge syndrome patient post-thymus transplantation.

Authors
Devlin, BH; Li, J; Alexieff, M; Harley, S; McCarthy, E; He, M; Kepler, TB; Markert, ML
MLA Citation
Devlin, BH, Li, J, Alexieff, M, Harley, S, McCarthy, E, He, M, Kepler, TB, and Markert, ML. "Time course of normalization of T cell receptor repertoire in a complete DiGeorge syndrome patient post-thymus transplantation." September 2005.
Source
wos-lite
Published In
Clinical Immunology
Volume
116
Issue
3
Publish Date
2005
Start Page
285
End Page
286

Statistical analysis of antigen receptor spectratype data.

MOTIVATION: The effectiveness of vertebrate adaptive immunity depends crucially on the establishment and maintenance of extreme diversity in the antigen receptor repertoire. Spectratype analysis is a method used in clinical and basic immunological settings in which antigen receptor length diversity is assessed as a surrogate for functional diversity. The purpose of this paper is to describe the systematic derivation and application of statistical methods for the analysis of spectratype data. RESULTS: The basic probability model used for spectratype analysis is the multinomial model with n, the total number of counts, indeterminate. We derive the appropriate statistics and statistical procedures for testing hypotheses regarding differences in antigen receptor distributions and variable repertoire diversity in different treatment groups. We then apply these methods to spectratype data obtained from several healthy donors to examine the differences between normal CD4+ and CD8+ T cell repertoires, and to data from a thymus transplant patient to examine the development of repertoire diversity following the transplant.

Authors
Kepler, TB; He, M; Tomfohr, JK; Devlin, BH; Sarzotti, M; Markert, ML
MLA Citation
Kepler, TB, He, M, Tomfohr, JK, Devlin, BH, Sarzotti, M, and Markert, ML. "Statistical analysis of antigen receptor spectratype data." Bioinformatics 21.16 (August 15, 2005): 3394-3400.
PMID
15955781
Source
pubmed
Published In
Bioinformatics
Volume
21
Issue
16
Publish Date
2005
Start Page
3394
End Page
3400
DOI
10.1093/bioinformatics/bti539

Thymic transplantation for complete DiGeorge syndrome: medical and surgical considerations.

BACKGROUND/PURPOSE: Complete DiGeorge syndrome results in the absence of functional T cells. Our program supports the transplantation of allogeneic thymic tissue in infants with DiGeorge syndrome to reconstitute immune function. This study reviews the multidisciplinary care of these complex infants. METHODS: From 1991 to 2001, the authors evaluated 16 infants with complete DiGeorge syndrome. All infants received multidisciplinary medical and surgical support. Clinical records for the group were reviewed. RESULTS: Four infants died without receiving a thymic transplantation, and 12 children survived to transplantation. The mean age at time of transplantation was 2.7 months (range, 1.1 to 4.4 months). All 16 infants had significant comorbidity including congenital heart disease (16 of 16), hypocalcemia (14 of 16), gastroesophageal reflux disease or aspiration (13 of 16), CHARGE complex (4 of 16), and other organ involvement (14 of 16). Nontransplant surgical procedures included central line placement (15 of 16), fundoplication or gastrostomy (10 of 16), cardiac repair (10 of 16), bronchoscopy or tracheostomy (6 of 16), and other procedures (12 of 16). Complications were substantial, and 5 of the 12 transplanted infants died of nontransplant-related conditions. All surviving infants have immune reconstitution, with follow-up from 2 to 10 years. CONCLUSIONS: Although the transplantation of thymic tissue can restore immune function in infants with complete DiGeorge syndrome, these children have substantial comorbidity. Care of these children requires coordinated multidisciplinary support.

Authors
Rice, HE; Skinner, MA; Mahaffey, SM; Oldham, KT; Ing, RJ; Hale, LP; Markert, ML
MLA Citation
Rice, HE, Skinner, MA, Mahaffey, SM, Oldham, KT, Ing, RJ, Hale, LP, and Markert, ML. "Thymic transplantation for complete DiGeorge syndrome: medical and surgical considerations." J Pediatr Surg 39.11 (November 2004): 1607-1615.
PMID
15547821
Source
pubmed
Published In
Journal of Pediatric Surgery
Volume
39
Issue
11
Publish Date
2004
Start Page
1607
End Page
1615

Postnatal thymus transplantation with immunosuppression as treatment for DiGeorge syndrome.

Complete DiGeorge syndrome is a fatal congenital disorder characterized by athymia, hypoparathyroidism, and heart defects. Less than half of patients are 22q11 hemizygous. The goal of this study was to assess if immune suppression followed by postnatal thymus transplantation would lead to T-cell function in 6 infant patients who had host T cells at the time of transplantation. All infants had fewer than 50 recent thymic emigrants (CD3(+)CD45RA(+)CD62L(+)) per cubic millimeter (mm(3)) and all had some proliferative response to the mitogen phytohemagglutinin. Four infants had rash, lymphadenopathy, and oligoclonal populations of T cells in the periphery. Five of 6 patients are alive at the follow-up interval of 15 months to 30 months. The 5 surviving patients developed a mean of 983 host CD3(+) T cells/mm(3) (range, 536/mm(3)-1574/mm(3)), a mean of 437 recent thymic emigrants/mm(3) (range, 196/mm(3)-785/mm(3)), and normal proliferative responses to phytohemaglutinin (follow-up from day 376 to day 873). The TCR repertoire became polyclonal in patients who presented with oligoclonal T cells. All patients had thymopoiesis on allograft biopsy. Postnatal thymus transplantation after treatment with Thymoglobulin shows promise as therapy for infants with complete DiGeorge syndrome who have significant proliferative responses to mitogens or who develop rash, lymphadenopathy, and oligoclonal T cells.

Authors
Markert, ML; Alexieff, MJ; Li, J; Sarzotti, M; Ozaki, DA; Devlin, BH; Sedlak, DA; Sempowski, GD; Hale, LP; Rice, HE; Mahaffey, SM; Skinner, MA
MLA Citation
Markert, ML, Alexieff, MJ, Li, J, Sarzotti, M, Ozaki, DA, Devlin, BH, Sedlak, DA, Sempowski, GD, Hale, LP, Rice, HE, Mahaffey, SM, and Skinner, MA. "Postnatal thymus transplantation with immunosuppression as treatment for DiGeorge syndrome." Blood 104.8 (October 15, 2004): 2574-2581.
PMID
15100156
Source
pubmed
Published In
Blood
Volume
104
Issue
8
Publish Date
2004
Start Page
2574
End Page
2581
DOI
10.1182/blood-2003-08-2984

Complete DiGeorge syndrome: development of rash, lymphadenopathy, and oligoclonal T cells in 5 cases.

BACKGROUND: Five patients with DiGeorge syndrome presented with infections, skin rashes, and lymphadenopathy after the newborn period. T-cell counts and function varied greatly in each patient. Initial laboratory testing did not suggest athymia in these patients. OBJECTIVE: The purpose of this study was to determine whether the patients had significant immunodeficiency. METHODS: Research testing of peripheral blood included immunoscope evaluation of T-cell receptor beta variable gene segment repertoire diversity, quantification of T-cell receptor rearrangement excision circles, and detection of naive T cells (expressing CD45RA and CD62L). RESULTS: The patients were classified as having DiGeorge syndrome on the basis of syndromic associations and heart, parathyroid, and immune abnormalities. Immunoscope evaluation revealed that the T-cell repertoires were strikingly oligoclonal in all patients. There were few recent thymic emigrants, as indicated by the very low numbers of naive T cells (<50/mm(3)) and the absence of T-cell receptor rearrangement excision circles. These studies showed that all 5 patients were athymic. Two patients died, one from infection. No thymus was found during the complete autopsy performed on one patient. CONCLUSION: Patients with DiGeorge syndrome, skin rash, and lymphadenopathy should undergo analysis of naive T-cell numbers and of T-cell receptor beta variability segment repertoire to determine whether they are athymic, even if they have T cells with mitogen responsiveness. It is important for physicians to realize that patients with complete DiGeorge syndrome remain profoundly immunodeficient after development of these atypical features (rash, lymphadenopathy, and oligoclonal T cells). Prompt diagnosis is necessary for appropriate management.

Authors
Markert, ML; Alexieff, MJ; Li, J; Sarzotti, M; Ozaki, DA; Devlin, BH; Sempowski, GD; Rhein, ME; Szabolcs, P; Hale, LP; Buckley, RH; Coyne, KE; Rice, HE; Mahaffey, SM; Skinner, MA
MLA Citation
Markert, ML, Alexieff, MJ, Li, J, Sarzotti, M, Ozaki, DA, Devlin, BH, Sempowski, GD, Rhein, ME, Szabolcs, P, Hale, LP, Buckley, RH, Coyne, KE, Rice, HE, Mahaffey, SM, and Skinner, MA. "Complete DiGeorge syndrome: development of rash, lymphadenopathy, and oligoclonal T cells in 5 cases." J Allergy Clin Immunol 113.4 (April 2004): 734-741.
PMID
15100681
Source
pubmed
Published In
Journal of Allergy and Clinical Immunology
Volume
113
Issue
4
Publish Date
2004
Start Page
734
End Page
741
DOI
10.1016/j.jaci.2004.01.766

T cell competition and repertoire diversity: the roles of TCR-specific and non-specific resource limitation determined through multivariate data analysis and computational modeling

Authors
Tomfohr, J; He, M; Cowell, L; Sarzotti-Kelsoe, M; Markert, ML; Kepler, TB
MLA Citation
Tomfohr, J, He, M, Cowell, L, Sarzotti-Kelsoe, M, Markert, ML, and Kepler, TB. "T cell competition and repertoire diversity: the roles of TCR-specific and non-specific resource limitation determined through multivariate data analysis and computational modeling." March 24, 2004.
Source
wos-lite
Published In
The FASEB journal : official publication of the Federation of American Societies for Experimental Biology
Volume
18
Issue
5
Publish Date
2004
Start Page
A798
End Page
A798

Statistical methods for quantitative analysis of TCR diversity via immunoscope

Authors
Kepler, TB; He, M; Tomfohr, J; Fadel, S; Sarzotti, M; Markert, ML
MLA Citation
Kepler, TB, He, M, Tomfohr, J, Fadel, S, Sarzotti, M, and Markert, ML. "Statistical methods for quantitative analysis of TCR diversity via immunoscope." March 24, 2004.
Source
wos-lite
Published In
The FASEB journal : official publication of the Federation of American Societies for Experimental Biology
Volume
18
Issue
5
Publish Date
2004
Start Page
A1181
End Page
A1181

Corticosteroids regulate epithelial cell differentiation and Hassall body formation in the human thymus.

The presence of characteristic epithelial swirls called Hassall bodies within the human thymic medulla has been used as an indicator of ongoing or recent thymopoiesis. We present a case where Hassall bodies were present in the absence of current or past thymopoiesis. The patient had been treated with corticosteroids for presumed asthma before his diagnosis of X-linked SCID. Two other cases of nonimmunodeficient patients treated with high-dose corticosteroids had markedly increased numbers of thymic Hassall bodies. To determine whether corticosteroid treatment induces thymic epithelial (TE) differentiation to form Hassall bodies, mAbs reactive with specific cytokeratins (CKs), filaggrin, and involucrin were used to define distinct stages of TE cell differentiation. Treatment of primary TE monolayers with hydrocortisone in vitro induced expression of involucrin and high-molecular-mass CKs that are characteristic of TE differentiation. Treatment of thymic organ cultures with hydrocortisone induced both medullary and subcapsular cortical TE cells to express CK6, a differentiation marker that is normally expressed only by Hassall bodies in vivo. These experimental studies combined with the case observations indicate that exogenous corticosteroids can regulate terminal differentiation of TE cells both in vitro and in vivo. Thus, the presence of Hassall bodies in thymus from corticosteroid-treated patients cannot be taken as an absolute indication of previous thymopoiesis. Because corticosteroids are also made within the thymus under normal physiologic conditions, these studies support the hypothesis that endogenous corticosteroids may play a role in normal TE differentiation and Hassall body formation in vivo.

Authors
Hale, LP; Markert, ML
MLA Citation
Hale, LP, and Markert, ML. "Corticosteroids regulate epithelial cell differentiation and Hassall body formation in the human thymus." J Immunol 172.1 (January 1, 2004): 617-624.
PMID
14688374
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
172
Issue
1
Publish Date
2004
Start Page
617
End Page
624

Thymus transplantation in complete DiGeorge syndrome: immunologic and safety evaluations in 12 patients.

Complete DiGeorge syndrome is a fatal condition in which infants have no detectable thymus function. The optimal treatment for the immune deficiency of complete DiGeorge syndrome has not been determined. Safety and efficacy of thymus transplantation were evaluated in 12 infants with complete DiGeorge syndrome who had less than 20-fold proliferative responses to phytohemagglutinin. All but one had fewer than 50 T cells/mm3. Allogeneic postnatal cultured thymus tissue was transplanted. T-cell development was followed by flow cytometry, lymphocyte proliferation assays, and T-cell receptor Vbeta (TCRBV) repertoire evaluation. Of the 12 patients, 7 are at home 15 months to 8.5 years after transplantation. All 7 survivors developed T-cell proliferative responses to mitogens of more than 100 000 counts per minute (cpm). By one year after transplantation, 6 of 7 patients developed antigen-specific proliferative responses. The TCRBV repertoire showed initial oligoclonality that progressed to polyclonality within a year. B-cell function developed in all 3 patients tested after 2 years. Deaths were associated with underlying congenital problems. Risk factors for death included tracheostomy, long-term mechanical ventilation, and cytomegalovirus infection. Adverse events in the first 3 months after transplantation included eosinophilia, rash, lymphadenopathy, development of CD4-CD8- peripheral T cells, elevated serum immunoglobulin E (IgE), and possible pulmonary inflammation. Adverse events related to the immune system occurring more than 3 months after transplantation included thrombocytopenia in one patient and hypothyroidism and alopecia in one other patient. Thymic transplantation is efficacious, well tolerated, and should be considered as treatment for infants with complete DiGeorge syndrome.

Authors
Markert, ML; Sarzotti, M; Ozaki, DA; Sempowski, GD; Rhein, ME; Hale, LP; Le Deist, F; Alexieff, MJ; Li, J; Hauser, ER; Haynes, BF; Rice, HE; Skinner, MA; Mahaffey, SM; Jaggers, J; Stein, LD; Mill, MR
MLA Citation
Markert, ML, Sarzotti, M, Ozaki, DA, Sempowski, GD, Rhein, ME, Hale, LP, Le Deist, F, Alexieff, MJ, Li, J, Hauser, ER, Haynes, BF, Rice, HE, Skinner, MA, Mahaffey, SM, Jaggers, J, Stein, LD, and Mill, MR. "Thymus transplantation in complete DiGeorge syndrome: immunologic and safety evaluations in 12 patients." Blood 102.3 (August 1, 2003): 1121-1130.
PMID
12702512
Source
pubmed
Published In
Blood
Volume
102
Issue
3
Publish Date
2003
Start Page
1121
End Page
1130
DOI
10.1182/blood-2002-08-2545

Thymus transplantation after immunosuppression with thymoglobulin.

Authors
Markert, ML; Sarzotti, M; Ozaki, DA; Hale, LP; Alexieff, MA; Li, J
MLA Citation
Markert, ML, Sarzotti, M, Ozaki, DA, Hale, LP, Alexieff, MA, and Li, J. "Thymus transplantation after immunosuppression with thymoglobulin." CLINICAL IMMUNOLOGY 103.3 (June 2002): S88-S88.
Source
wos-lite
Published In
Clinical Immunology
Volume
103
Issue
3
Publish Date
2002
Start Page
S88
End Page
S88

U.S. Department of Health and Human Services National Institutes of Health Recombinant DNA Advisory Committee: Minutes of meeting: December 6, 2001

Authors
Aguilar-Cordova, CE; Macklin, R; Markert, ML; Mickelson, CA; Ando, DG; Breakefield, XO; Friedmann, TC; Gordon, JW; Greenblatt, JJ; Juengst, ET; King, NMP; Levi-Pearl, SL
MLA Citation
Aguilar-Cordova, CE, Macklin, R, Markert, ML, Mickelson, CA, Ando, DG, Breakefield, XO, Friedmann, TC, Gordon, JW, Greenblatt, JJ, Juengst, ET, King, NMP, and Levi-Pearl, SL. "U.S. Department of Health and Human Services National Institutes of Health Recombinant DNA Advisory Committee: Minutes of meeting: December 6, 2001." Human Gene Therapy 13.13 (2002): 1663-1673.
Source
scival
Published In
Human Gene Therapy
Volume
13
Issue
13
Publish Date
2002
Start Page
1663
End Page
1673
DOI
10.1089/10430340260201752

Thymopoiesis in HIV-infected adults after highly active antiretroviral therapy.

The thymus of HIV-seropositive patients can enlarge as CD4+ T cell counts increase on highly active anti-retroviral therapy (HAART). This may indicate development of new T cells or represent mature peripheral T cells recirculating to the thymus. To define the etiology of the enlargement, the thymuses of two HIV-infected individuals on HAART were biopsied. For more than 3 years before initiation of HAART, both patients (38 and 41 years of age) had documented CD4+ T lymphopenia. Peripheral blood samples were obtained to assess circulating CD4+ CD45RA+ CD62L+ T cells, which were thought to have recently developed in the thymus. Peripheral blood T cells from both patients and thymocytes from the second patient were also tested for levels of DNA episomes formed during T cell receptor gene rearrangement (T cell receptor rearrangement excision circles, TRECs). With HAART, peripheral blood CD4+ T cell counts increased from approximately 60/mm(3) to 552/mm(3) and 750/mm(3) for patients 1 and 2, respectively. Thymic biopsies from both patients showed normal thymus histology with active thymopoiesis. Percentages of peripheral blood CD4+ CD45RA+ CD62L+ T cells and quantitation of T cell TRECs also reflected active thymopoiesis in both patients. Thus, in these two HIV-seropositive adults examined after initiation of HAART, thymic enlargement represented active thymopoiesis. Thymopoiesis in adult AIDS patients may contribute to immune reconstitution even after prolonged CD4+ T lymphopenia.

Authors
Markert, ML; Alvarez-McLeod, AP; Sempowski, GD; Hale, LP; Horvatinovich, JM; Weinhold, KJ; Bartlett, JA; D'Amico, TA; Haynes, BF
MLA Citation
Markert, ML, Alvarez-McLeod, AP, Sempowski, GD, Hale, LP, Horvatinovich, JM, Weinhold, KJ, Bartlett, JA, D'Amico, TA, and Haynes, BF. "Thymopoiesis in HIV-infected adults after highly active antiretroviral therapy." AIDS Res Hum Retroviruses 17.17 (November 20, 2001): 1635-1643.
PMID
11779351
Source
pubmed
Published In
AIDS Research and Human Retroviruses
Volume
17
Issue
17
Publish Date
2001
Start Page
1635
End Page
1643
DOI
10.1089/088922201753342040

Prenatal gene tranfer: scientific, medical, and ethical issues: a report of the Recombinant DNA Advisory Committee.

MLA Citation
"Prenatal gene tranfer: scientific, medical, and ethical issues: a report of the Recombinant DNA Advisory Committee." Human gene therapy 11.8 (May 2000): 1211-1229.
PMID
11660775
Source
epmc
Published In
Human Gene Therapy
Volume
11
Issue
8
Publish Date
2000
Start Page
1211
End Page
1229
DOI
10.1089/10430340050015257

Effect of highly active antiretroviral therapy and thymic transplantation on immunoreconstitution in HIV infection.

The purpose of this study was to determine whether thymic transplantation in addition to highly active antiretroviral therapy (HAART) will restore T cell function in HIV infection. Eight treatment-naive HIV-infected patients with CD4+ T cell counts of 200-500/mm3 were randomized into thymic transplantation and control arms. All patients received HAART (zidovudine, lamivudine, and ritonavir) for 6 weeks prior to transplantation. Thymic transplantation was done without immunosuppression, using postnatal HLA-unmatched cultured allogeneic thymus tissue. Patients were immunized every 6 months with the neoantigen keyhole limpet hemocyanin (KLH) and the recall antigen tetanus toxoid (TT). T cell phenotype and function and T cell receptor rearrangement excision circles (TRECs) were assessed. Thymic allografts were biopsied at 2 months. Six HIV-infected patients completed the study. Four patients received cultured allogeneic postnatal thymic grafts, two others were controls. CD4+ T cell counts increased and T cell-proliferative responses to Candida antigen and TT normalized in all patients. Proliferative responses to KLH developed in three of four transplant recipients and one of two controls. Patients responding to KLH after secondary immunization had greater TREC increases compared with the patients who did not respond. All thymic allografts were rejected within 2 months. In summary, four of six patients developed T cell-proliferative responses to the neoantigen KLH over the first 2 years of HAART. The transplanted thymus tissue, however, was rejected. There was no clear difference in restoration of T cell function in the transplant recipients compared with the controls. Increases in TRECs after initiation of HAART may correlate with improved immune function.

Authors
Markert, ML; Hicks, CB; Bartlett, JA; Harmon, JL; Hale, LP; Greenberg, ML; Ferrari, G; Ottinger, J; Boeck, A; Kloster, AL; McLaughlin, TM; Bleich, KB; Ungerleider, RM; Lyerly, HK; Wilkinson, WE; Rousseau, FS; Heath-Chiozzi, ME; Leonard, JM; Haase, AT; Shaw, GM; Bucy, RP; Douek, DC; Koup, RA; Haynes, BF; Bolognesi, DP; Weinhold, KJ
MLA Citation
Markert, ML, Hicks, CB, Bartlett, JA, Harmon, JL, Hale, LP, Greenberg, ML, Ferrari, G, Ottinger, J, Boeck, A, Kloster, AL, McLaughlin, TM, Bleich, KB, Ungerleider, RM, Lyerly, HK, Wilkinson, WE, Rousseau, FS, Heath-Chiozzi, ME, Leonard, JM, Haase, AT, Shaw, GM, Bucy, RP, Douek, DC, Koup, RA, Haynes, BF, Bolognesi, DP, and Weinhold, KJ. "Effect of highly active antiretroviral therapy and thymic transplantation on immunoreconstitution in HIV infection." AIDS Res Hum Retroviruses 16.5 (March 20, 2000): 403-413.
PMID
10772526
Source
pubmed
Published In
AIDS Research and Human Retroviruses
Volume
16
Issue
5
Publish Date
2000
Start Page
403
End Page
413
DOI
10.1089/088922200309061

The role of the thymus in immune reconstitution in aging, bone marrow transplantation, and HIV-1 infection.

The human thymus is a complex chimeric organ comprised of central (thymic epithelial space) and peripheral (perivascular space) components that functions well into adult life to produce naive T lymphocytes. Recent advances in identifying thymic emigrants and development of safe methods to study thymic function in vivo in adults have provided new opportunities to understand the role that the human thymus plays in immune reconstitution in aging, in bone marrow transplantation, and in HIV-1 infection. The emerging concept is that there are age-dependent contributions of thymic emigrants and proliferation of postthymic T cells to maintain the peripheral T cell pool and to contribute to T cell regeneration, with the thymus contributing more at younger ages and peripheral T cell expansion contributing more in older subjects. New studies have revealed a dynamic interplay between postnatal thymus output and peripheral T cell pool proliferation, which play important roles in determining the nature of immune reconstitution in congenital immunodeficiency diseases, in bone marrow transplantation, and in HIV-1 infection. In this paper, we review recent data on human postnatal thymus function that, taken together, support the notion that the human thymus is functional well into the sixth decade and plays a role throughout life to optimize human immune system function.

Authors
Haynes, BF; Markert, ML; Sempowski, GD; Patel, DD; Hale, LP
MLA Citation
Haynes, BF, Markert, ML, Sempowski, GD, Patel, DD, and Hale, LP. "The role of the thymus in immune reconstitution in aging, bone marrow transplantation, and HIV-1 infection." Annu Rev Immunol 18 (2000): 529-560. (Review)
PMID
10837068
Source
pubmed
Published In
Annual Review of Immunology
Volume
18
Publish Date
2000
Start Page
529
End Page
560
DOI
10.1146/annurev.immunol.18.1.529

Department of health and human services national institutes of health recombinant DNA advisory committee: Minutes of meeting March 8-10, 2000

Authors
Aguilar-Cordova, CE; Levi-Pearl, SL; Macklin, R; Markert, ML; McIvor, RS; Mickelson, CA; Wolff, JA; Ando, DG; Breakefield, XO; Chow, LT; Friedmann, T; Gordon, JW; Greenblatt, JJ; Juengst, ET; King, NMP
MLA Citation
Aguilar-Cordova, CE, Levi-Pearl, SL, Macklin, R, Markert, ML, McIvor, RS, Mickelson, CA, Wolff, JA, Ando, DG, Breakefield, XO, Chow, LT, Friedmann, T, Gordon, JW, Greenblatt, JJ, Juengst, ET, and King, NMP. "Department of health and human services national institutes of health recombinant DNA advisory committee: Minutes of meeting March 8-10, 2000." Human Gene Therapy 11.15 (2000): 2159-2192.
PMID
11044916
Source
scival
Published In
Human Gene Therapy
Volume
11
Issue
15
Publish Date
2000
Start Page
2159
End Page
2192
DOI
10.1089/104303400750001453

Transplantation of thymus tissue in complete DiGeorge syndrome.

BACKGROUND: The DiGeorge syndrome is a congenital disorder that affects the heart, parathyroid glands, and thymus. In complete DiGeorge syndrome, patients have severely reduced T-cell function. METHODS: We treated five infants (age, one to four months) with complete DiGeorge syndrome by transplantation of cultured postnatal thymus tissue. Follow-up evaluations included immune phenotyping and proliferative studies of peripheral-blood mononuclear cells plus biopsy of the thymus allograft. Thymic production of new T cells was assessed in peripheral blood by tests for T-cell-receptor recombination excision circles, which are formed from excised DNA during the rearrangement of T-cell-receptor genes. RESULTS: After the transplantation of thymus tissue, T-cell proliferative responses to mitogens developed in four of the five patients. Two of the patients survived with restoration of immune function; three patients died from infection or abnormalities unrelated to transplantation. Biopsies of grafted thymus in the surviving patients showed normal morphologic features and active T-cell production. In three patients, donor T cells could be detected about four weeks after transplantation, although there was no evidence of graft-versus-host disease on biopsy or at autopsy. In one patient, the T-cell development within the graft was demonstrated to accompany the appearance of recently developed T cells in the periphery and coincided with the onset of normal T-cell function. In one patient, there was evidence of thymus function and CD45RA+CD62L+ T cells more than five years after transplantation. CONCLUSIONS: In some infants with profound immunodeficiency and complete DiGeorge syndrome, the transplantation of thymus tissue can restore normal immune function. Early thymus transplantation - before the development of infectious complications - may promote successful immune reconstitution.

Authors
Markert, ML; Boeck, A; Hale, LP; Kloster, AL; McLaughlin, TM; Batchvarova, MN; Douek, DC; Koup, RA; Kostyu, DD; Ward, FE; Rice, HE; Mahaffey, SM; Schiff, SE; Buckley, RH; Haynes, BF
MLA Citation
Markert, ML, Boeck, A, Hale, LP, Kloster, AL, McLaughlin, TM, Batchvarova, MN, Douek, DC, Koup, RA, Kostyu, DD, Ward, FE, Rice, HE, Mahaffey, SM, Schiff, SE, Buckley, RH, and Haynes, BF. "Transplantation of thymus tissue in complete DiGeorge syndrome." N Engl J Med 341.16 (October 14, 1999): 1180-1189.
PMID
10523153
Source
pubmed
Published In
The New England journal of medicine
Volume
341
Issue
16
Publish Date
1999
Start Page
1180
End Page
1189
DOI
10.1056/NEJM199910143411603

Possible extrathymic development of nonfunctional T cells in a patient with complete DiGeorge syndrome.

Complete DiGeorge syndrome is characterized by the clinical triad of cardiac malformation, hypocalcemia, and T cell immunodeficiency due to congenital athymia. We describe an infant with complete DiGeorge syndrome who at presentation had no circulating T cells detectable by flow cytometry. The patient spontaneously developed circulating T cells but these cells did not proliferate in response to mitogens. The T cell receptor Vbeta repertoire was severely restricted. All T cells were host, not maternal, as assessed by fluorescent in situ hybridization evaluation of 22q11 hemizygosity. At autopsy, this patient had no grossly detectable thymus tissue and no microscopic evidence for thymopoiesis. These findings suggest that appearance of T cells in infants with complete DiGeorge syndrome may represent oligoclonal expansions of a small number of T cells that may have matured extrathymically and which do not respond in vitro to mitogen stimulation.

Authors
Collard, HR; Boeck, A; Mc Laughlin, TM; Watson, TJ; Schiff, SE; Hale, LP; Markert, ML
MLA Citation
Collard, HR, Boeck, A, Mc Laughlin, TM, Watson, TJ, Schiff, SE, Hale, LP, and Markert, ML. "Possible extrathymic development of nonfunctional T cells in a patient with complete DiGeorge syndrome." Clin Immunol 91.2 (May 1999): 156-162.
PMID
10227807
Source
pubmed
Published In
Clinical Immunology
Volume
91
Issue
2
Publish Date
1999
Start Page
156
End Page
162
DOI
10.1006/clim.1999.4691

Hematopoietic stem-cell transplantation for the treatment of severe combined immunodeficiency.

BACKGROUND: Since 1968 it has been known that bone marrow transplantation can ameliorate severe combined immunodeficiency, but data on the long-term efficacy of this treatment are limited. We prospectively studied immunologic function in 89 consecutive infants with severe combined immunodeficiency who received hematopoietic stem-cell transplants at Duke University Medical Center between May 1982 and September 1998. METHODS: Serum immunoglobulin levels and lymphocyte phenotypes and function were assessed and genetic analyses performed according to standard methods. Bone marrow was depleted of T cells by agglutination with soybean lectin and by sheep-erythrocyte rosetting before transplantation. RESULTS: Seventy-seven of the infants received T-cell-depleted, HLA-haploidentical parental marrow, and 12 received HLA-identical marrow from a related donor; 3 of the recipients of haploidentical marrow also received placental-blood transplants from unrelated donors. Except for two patients who received placental blood, none of the recipients received chemotherapy before transplantation or prophylaxis against graft-versus-host disease. Of the 89 infants, 72 (81 percent) were still alive 3 months to 16.5 years after transplantation, including all of the 12 who received HLA-identical marrow, 60 of the 77 (78 percent) who were given haploidentical marrow, and 2 of the 3 (67 percent) who received both haploidentical marrow and placental blood. T-cell function became normal within two weeks after transplantation in the patients who received unfractionated HLA-identical marrow but usually not until three to four months after transplantation in those who received T-cell-depleted marrow. At the time of the most recent evaluation, all but 4 of the 72 survivors had normal T-cell function, and all the T cells in their blood were of donor origin. B-cell function remained abnormal in many of the recipients of haploidentical marrow. In 26 children (5 recipients of HLA-identical marrow and 21 recipients of haploidentical marrow) between 2 percent and 100 percent of B cells were of donor origin. Forty-five of the 72 children were receiving intravenous immune globulin. CONCLUSIONS: Transplantation of marrow from a related donor is a life-saving and life-sustaining treatment for patients with any type of severe combined immunodeficiency, even when there is no HLA-identical donor.

Authors
Buckley, RH; Schiff, SE; Schiff, RI; Markert, L; Williams, LW; Roberts, JL; Myers, LA; Ward, FE
MLA Citation
Buckley, RH, Schiff, SE, Schiff, RI, Markert, L, Williams, LW, Roberts, JL, Myers, LA, and Ward, FE. "Hematopoietic stem-cell transplantation for the treatment of severe combined immunodeficiency." N Engl J Med 340.7 (February 18, 1999): 508-516.
PMID
10021471
Source
pubmed
Published In
The New England journal of medicine
Volume
340
Issue
7
Publish Date
1999
Start Page
508
End Page
516
DOI
10.1056/NEJM199902183400703

Complete DiGeorge syndrome: persistence of profound immunodeficiency.

OBJECTIVE: DiGeorge syndrome is characterized by developmental defects of the heart, parathyroid glands, and thymus. The objective of this study was to determine whether T-cell function spontaneously improves in patients with DiGeorge syndrome who have profoundly depressed T-cell proliferative responses to mitogens at presentation, regardless of the T-cell count. STUDY DESIGN: We conducted a retrospective chart review of eight patients with DiGeorge syndrome who had no proliferative responses to mitogens on presentation. RESULTS: Despite lack of responsiveness of the patients' peripheral blood lymphocytes to mitogens, T cells were occasionally detected, and the patients' cells often responded to IL-2 and in mixed lymphocyte reactions. Unresponsiveness to mitogens and clinical immunodeficiency persisted without immune-based therapy. One patient is alive and well after immunoreconstitution from thymic transplantation. The others either died early of complications of their disease such as gastroesophageal reflux with aspiration (2 patients) or infection (2 patients) or died after attempts at immunorestorative therapy with IL-2, thymus transplantation, or bone marrow transplantation (3 patients). CONCLUSION: Eight patients with DiGeorge syndrome who were first seen with no mitogen responsiveness did not improve spontaneously. We recommend HLA-identical bone marrow transplantation or thymic transplantation for these patients as soon as the diagnosis is confirmed.

Authors
Markert, ML; Hummell, DS; Rosenblatt, HM; Schiff, SE; Harville, TO; Williams, LW; Schiff, RI; Buckley, RH
MLA Citation
Markert, ML, Hummell, DS, Rosenblatt, HM, Schiff, SE, Harville, TO, Williams, LW, Schiff, RI, and Buckley, RH. "Complete DiGeorge syndrome: persistence of profound immunodeficiency." J Pediatr 132.1 (January 1998): 15-21.
PMID
9469994
Source
pubmed
Published In
The Journal of Pediatrics
Volume
132
Issue
1
Publish Date
1998
Start Page
15
End Page
21

Recombinant DNA Advisory Committee Meeting, National Institutes of Health, 9000 Rockville Pike, Bethesda, Maryland 20892, March 10 1998: Department of Health and Human Services - National Institutes of Health

Authors
Mickelson, CA; Gordon, JW; Markert, ML; Greenblatt, JJ; McIvor, RS; Aguilar-Codova, CE; Kumar, R; Tufaro, F; Osborne, S; Skirboll, ; Macklin, R; Fost, ; Lantos, ; Taylor, A
MLA Citation
Mickelson, CA, Gordon, JW, Markert, ML, Greenblatt, JJ, McIvor, RS, Aguilar-Codova, CE, Kumar, R, Tufaro, F, Osborne, S, Skirboll, , Macklin, R, Fost, , Lantos, , and Taylor, A. "Recombinant DNA Advisory Committee Meeting, National Institutes of Health, 9000 Rockville Pike, Bethesda, Maryland 20892, March 10 1998: Department of Health and Human Services - National Institutes of Health." Human Gene Therapy 9.16 (1998): 2427-2454.
PMID
9890758
Source
scival
Published In
Human Gene Therapy
Volume
9
Issue
16
Publish Date
1998
Start Page
2427
End Page
2454

Regulatory issues. Recombinant DNA advisory committee, department of health and human services, national institutes of health. Minutes of meeting December 15-16, 1997

Authors
Aguilar-Cordova, CE; Ando, DG; Gordon, JW; Greenblatt, JJ; Lai, MMC; Leinwand, LA; Lysaught, MT; Macklin, R; Markert, ML; McIvor, RS; Mickelson, CA; Rothenberg, K; Verma, IM; Wolff, JA; Knorr, DW; Jr, DFW; Jones, DD; Noguchi, P
MLA Citation
Aguilar-Cordova, CE, Ando, DG, Gordon, JW, Greenblatt, JJ, Lai, MMC, Leinwand, LA, Lysaught, MT, Macklin, R, Markert, ML, McIvor, RS, Mickelson, CA, Rothenberg, K, Verma, IM, Wolff, JA, Knorr, DW, Jr, DFW, Jones, DD, and Noguchi, P. "Regulatory issues. Recombinant DNA advisory committee, department of health and human services, national institutes of health. Minutes of meeting December 15-16, 1997." Human Gene Therapy 9.11 (1998): 1643-1684.
PMID
9694162
Source
scival
Published In
Human Gene Therapy
Volume
9
Issue
11
Publish Date
1998
Start Page
1643
End Page
1684

Severe laryngomalacia and bronchomalacia in DiGeorge syndrome and CHARGE association.

Authors
Markert, ML; Majure, M; Harville, TO; Hulka, G; Oldham, K
MLA Citation
Markert, ML, Majure, M, Harville, TO, Hulka, G, and Oldham, K. "Severe laryngomalacia and bronchomalacia in DiGeorge syndrome and CHARGE association." Pediatr Pulmonol 24.5 (November 1997): 364-369.
PMID
9407570
Source
pubmed
Published In
Pediatric Pulmonology
Volume
24
Issue
5
Publish Date
1997
Start Page
364
End Page
369

Human severe combined immunodeficiency: genetic, phenotypic, and functional diversity in one hundred eight infants.

OBJECTIVE: To determine the relative frequencies of the different genetic forms of severe combined immunodeficiency (SCID) and whether there are distinctive characteristics of the particular genotypes. STUDY DESIGN: The demographic, genetic, and immunologic features of 108 infants with SCID who were treated consecutively at Duke University Medical Center were analyzed. RESULTS: Eighty-nine subjects were boys and 19 were girls; there were 84 white infants, 16 black infants, and 8 Hispanic infants. Forty-nine had X-linked SCID with mutations of common cytokine receptor gamma chain (gamma c), 16 had adenosine deaminase (ADA) deficiency, 8 had Janus kinase 3 (Jak3) deficiency, 21 had unknown autosomal recessive mutations, 1 had reticular dysgenesis, 1 had cartilage hair hypoplasia, and 12 (all boys) had SCID of undetermined type. Deficiency of ADA caused the most profound lymphopenia; gamma c or Jak3 deficiency resulted in the most B cells and fewest natural killer (NK) cells; NK cells and function were highest in autosomal recessive and unknown types of SCID. CONCLUSIONS: Different SCID genotypes are associated with distinctive lymphocyte characteristics. The presence of NK function in ADA-deficient, autosomal recessive, and unknown type SCIDs, and low NK function in a majority of gamma c and Jak3 SCIDs indicates that some molecular lesions affect T, B, and NK cells (gamma c and Jak3), others primarily T cells (ADA deficiency), and others just T and B cells.

Authors
Buckley, RH; Schiff, RI; Schiff, SE; Markert, ML; Williams, LW; Harville, TO; Roberts, JL; Puck, JM
MLA Citation
Buckley, RH, Schiff, RI, Schiff, SE, Markert, ML, Williams, LW, Harville, TO, Roberts, JL, and Puck, JM. "Human severe combined immunodeficiency: genetic, phenotypic, and functional diversity in one hundred eight infants." J Pediatr 130.3 (March 1997): 378-387.
PMID
9063412
Source
pubmed
Published In
The Journal of Pediatrics
Volume
130
Issue
3
Publish Date
1997
Start Page
378
End Page
387

Normalization of the peripheral blood T cell receptor V beta repertoire after cultured postnatal human thymic transplantation in DiGeorge syndrome.

Complete DiGeorge syndrome is an immunodeficiency disease characterized by thymic aplasia and the absence of functioning peripheral T cells. A patient with this syndrome was transplanted with cultured postnatal human thymic tissue. Within 5 weeks of transplantation, flow cytometry, T cell receptor V beta sequence analysis, and cell function studies showed the presence of oligoclonal populations of nonfunctional clonally expanded peripheral T cells that were derived from pretransplantation T cells present in the skin. However, at 3 months posttransplantation, a biopsy of the transplanted thymus showed normal intrathymic T cell maturation of host T cells with normal TCR V beta expression on thymocytes. By 9 months postransplantation, peripheral T cell function was restored and the TCR V beta repertoire became polyclonal, coincident with the appearance of normal T cell function. These data suggest that the transplanted thymus was responsible for the establishment of a new T cell repertoire via thymopoiesis in the chimeric thymic graft.

Authors
Davis, CM; McLaughlin, TM; Watson, TJ; Buckley, RH; Schiff, SE; Hale, LP; Haynes, BF; Markert, ML
MLA Citation
Davis, CM, McLaughlin, TM, Watson, TJ, Buckley, RH, Schiff, SE, Hale, LP, Haynes, BF, and Markert, ML. "Normalization of the peripheral blood T cell receptor V beta repertoire after cultured postnatal human thymic transplantation in DiGeorge syndrome." J Clin Immunol 17.2 (March 1997): 167-175.
PMID
9083893
Source
pubmed
Published In
Journal of Clinical Immunology
Volume
17
Issue
2
Publish Date
1997
Start Page
167
End Page
175

Successful formation of a chimeric human thymus allograft following transplantation of cultured postnatal human thymus.

Transplantation of cultured postnatal human thymus was performed in a patient with complete DiGeorge syndrome. Biopsy of the graft 3 mo after implantation revealed normal CD1+ thymocytes in thymic cortical epithelial regions and CD1- thymocytes in thymic medullary epithelial regions, respectively. HLA analysis of graft thymocyte and thymic microenvironment components demonstrated that developing thymocytes and thymic macrophages were recipient derived, while thymic epithelial components were of donor origin. The patient, who initially had no T cells and had profoundly defective T cell function, developed normal T cell responses to mitogens and Ags, tolerance to donor in a mixed lymphocyte reaction, and normal Ab titers after tetanus toxoid and pneumovax immunization. Thus, transplantation of cultured postnatal human thymic tissue in humans can form functional chimeric thymic tissue, and may provide a strategy to reconstitute the peripheral T cell pool in select congenital and acquired immune deficiency syndromes.

Authors
Markert, ML; Kostyu, DD; Ward, FE; McLaughlin, TM; Watson, TJ; Buckley, RH; Schiff, SE; Ungerleider, RM; Gaynor, JW; Oldham, KT; Mahaffey, SM; Ballow, M; Driscoll, DA; Hale, LP; Haynes, BF
MLA Citation
Markert, ML, Kostyu, DD, Ward, FE, McLaughlin, TM, Watson, TJ, Buckley, RH, Schiff, SE, Ungerleider, RM, Gaynor, JW, Oldham, KT, Mahaffey, SM, Ballow, M, Driscoll, DA, Hale, LP, and Haynes, BF. "Successful formation of a chimeric human thymus allograft following transplantation of cultured postnatal human thymus." J Immunol 158.2 (January 15, 1997): 998-1005.
PMID
8993022
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
158
Issue
2
Publish Date
1997
Start Page
998
End Page
1005

The human thymic microenvironment during organ culture.

Cultured human thymic tissue has been transplanted into many patients with T cell dysfunction; however, little is known about the effect of in vitro culture on thymic tissue. Human postnatal thymic organ cultures were established in vitro to study the growth potential of the thymic epithelium and the expression of intracellular and surface antigens with time in culture. Marked depletion of bone marrow-derived cells was observed within 3 weeks of initiation of organ cultures although some viable CD3+ cells could still be detected. Thymic epithelial cells in in vitro explants continued to express MHC class I and class II antigens as well as cytokeratins. Thymic epithelial cells within cultured thymic organ slices maintained their postnatal growth potential, in that cytokeratin-positive epithelial monolayers could be established in vitro from these thymic slices up to 12 weeks after initiation of organ culture. Thus, thymic explants remained viable in culture and could potentially be used to reconstitute immunity in T cell deficient patients.

Authors
Markert, ML; Watson, TJ; Kaplan, I; Hale, LP; Haynes, BF
MLA Citation
Markert, ML, Watson, TJ, Kaplan, I, Hale, LP, and Haynes, BF. "The human thymic microenvironment during organ culture." Clin Immunol Immunopathol 82.1 (January 1997): 26-36.
PMID
9000039
Source
pubmed
Published In
Clinical Immunology and Immunopathology
Volume
82
Issue
1
Publish Date
1997
Start Page
26
End Page
36

Mutations in purine nucleoside phosphorylase deficiency.

Purine nucleoside phosphorylase deficiency is an inherited disease of purine metabolism characterized clinically as combined immunodeficiency. The molecular defects have been published for 4 different alleles in 3 patients. We report four new mutations including two amino acid substitutions, A174P and G190V, a single codon deletion, delta I129, and a point mutation in intron 3 which leads to aberrant splicing and creation of a premature stop codon in exon 4 (286-18G-->A). Of the previously reported mutations, E89K was found in one additional patient, and R234P was found in 3 unrelated patients, making R234P the most common mutation reported to date in this disease.

Authors
Markert, ML; Finkel, BD; McLaughlin, TM; Watson, TJ; Collard, HR; McMahon, CP; Andrews, LG; Barrett, MJ; Ward, FE
MLA Citation
Markert, ML, Finkel, BD, McLaughlin, TM, Watson, TJ, Collard, HR, McMahon, CP, Andrews, LG, Barrett, MJ, and Ward, FE. "Mutations in purine nucleoside phosphorylase deficiency." Hum Mutat 9.2 (1997): 118-121.
PMID
9067751
Source
pubmed
Published In
Human Mutation
Volume
9
Issue
2
Publish Date
1997
Start Page
118
End Page
121
DOI
10.1002/(SICI)1098-1004(1997)9:2<118::AID-HUMU3>3.0.CO;2-5

Antibodies against retroviral proteins and nuclear antigens in a subset of idiopathic CD4+ T lymphocytopenia patients.

Idiopathic CD4+ T lymphocytopenia (ICL) is an immunodeficiency syndrome characterized by severe depletion of CD4+ T lymphocytes, but in which human immunodeficiency virus cannot be detected. Peripheral blood mononuclear cells (BPMCs) from an ICL patient were cocultured with HUT78 T-lymphoblastoid cells, and an acute cytopathic effect and formation of multinucleated cells were observed. A human intracisternal A-type retroviral particle designated HIAP-II was detected in cells surviving the acute cytopathic effect. Eight of 13 ICL patients in a blinded screen of a serological panel provided by the National Centers for Disease Control and Prevention (CDC) had serum antibodies that specifically reacted with HIAP-II associated proteins by Western immunoblotting. None of 19 control sera in the panel that were unreactive with HIV Gag proteins produced a positive result on HIAP-II immunoblots. Comparable results were obtained in a blinded screen of a second CDC serological panel. Sera from 8 of 14 ICL patients in the second serological panel were positive for antinuclear autoantibodies (ANAs) commonly observed in patients with systemic autoimmune diseases. These results suggest the possible involvement of an A-type retrovirus or autoimmunity in development of ICL in a subset of patients.

Authors
Garry, RF; Fermin, CD; Kohler, PF; Markert, ML; Luo, H
MLA Citation
Garry, RF, Fermin, CD, Kohler, PF, Markert, ML, and Luo, H. "Antibodies against retroviral proteins and nuclear antigens in a subset of idiopathic CD4+ T lymphocytopenia patients." AIDS Res Hum Retroviruses 12.10 (July 1, 1996): 931-940.
PMID
8798978
Source
pubmed
Published In
AIDS Research and Human Retroviruses
Volume
12
Issue
10
Publish Date
1996
Start Page
931
End Page
940
DOI
10.1089/aid.1996.12.931

Perspective: research highlights at the Duke University center for AIDS research. Immunoreconstitution in HIV infection: the role of the thymus.

Authors
Markert, ML
MLA Citation
Markert, ML. "Perspective: research highlights at the Duke University center for AIDS research. Immunoreconstitution in HIV infection: the role of the thymus." AIDS Res Hum Retroviruses 12.9 (June 10, 1996): 751-755. (Review)
PMID
8738425
Source
pubmed
Published In
AIDS Research and Human Retroviruses
Volume
12
Issue
9
Publish Date
1996
Start Page
751
End Page
755
DOI
10.1089/aid.1996.12.751

Immunoreconstitution in HIV infection: The role of the thymus

Authors
Markert, ML
MLA Citation
Markert, ML. "Immunoreconstitution in HIV infection: The role of the thymus." AIDS RESEARCH AND HUMAN RETROVIRUSES 12.9 (June 10, 1996): 751-755.
Source
wos-lite
Published In
AIDS Research and Human Retroviruses
Volume
12
Issue
9
Publish Date
1996
Start Page
751
End Page
755
DOI
10.1089/aid.1996.12.751

Thymic transplantation for digeorge syndrome

Infants with complete DiGeorge syndrome have thymic aplasia with no measurable T cell function. This report describes restoration of T cell function and TCR repertoire during a 30 month period after thymic transplantation in a patient with complete DiGeorge syndrome. T cell function developed beginning at 7 months after transplantation. Currently, peripheral blood mononuclear cell (PBMNC) responses to PHA, ConA, and PWM, are normal. The patient's PBMNC displayed normal reactivity in mixed lymphocyte cultures but were tolerant of the thymic. TCR V repertoire studies (by flow cytometry and RT PCR and sequencing) showed an early clonal expansion of T cells and a later normalization of TCR repertoire. The patient developed normal serum immunoglobulin levels and normal antibody responses to tetanus toxoid and pneumococcus serotype 3. The patient is clinically well on supplemental calcium. Thymic transplantation should be considered in patients with complete DiGeorge syndrome.

Authors
Markert, ML
MLA Citation
Markert, ML. "Thymic transplantation for digeorge syndrome." FASEB Journal 10.6 (1996): A1063-.
Source
scival
Published In
The FASEB journal : official publication of the Federation of American Societies for Experimental Biology
Volume
10
Issue
6
Publish Date
1996
Start Page
A1063

Mutation of Jak3 in a patient with SCID: essential role of Jak3 in lymphoid development.

Males with X-linked severe combined immunodeficiency (XSCID) have defects in the common cytokine receptor gamma chain (gamma c) gene that encodes a shared, essential component of the receptors of interleukin-2 (IL-2), IL-4, IL-7, IL-9, and IL-15. The Janus family tyrosine kinase Jak3 is the only signaling molecule known to be associated with gamma c, so it was hypothesized that defects in Jak3 might cause an XSCID-like phenotype. A girl with immunological features indistinguishable from those of XSCID was therefore selected for analysis. An Epstein-Barr virus (EBV)-transformed cell line derived from her lymphocytes had normal gamma c expression but lacked Jak3 protein and had greatly diminished Jak3 messenger RNA. Sequencing revealed a different mutation on each allele: a single nucleotide insertion resulting in a frame shift and premature termination in the Jak3 JH4 domain and a nonsense mutation in the Jak3 JH2 domain. The lack of Jak3 expression correlated with impaired B cell signaling, as demonstrated by the inability of IL-4 to activate Stat6 in the EBV-transformed cell line from the patient. These observations indicate that the functions of gamma c are dependent on Jak3 and that Jak3 is essential for lymphoid development and signaling.

Authors
Russell, SM; Tayebi, N; Nakajima, H; Riedy, MC; Roberts, JL; Aman, MJ; Migone, TS; Noguchi, M; Markert, ML; Buckley, RH; O'Shea, JJ; Leonard, WJ
MLA Citation
Russell, SM, Tayebi, N, Nakajima, H, Riedy, MC, Roberts, JL, Aman, MJ, Migone, TS, Noguchi, M, Markert, ML, Buckley, RH, O'Shea, JJ, and Leonard, WJ. "Mutation of Jak3 in a patient with SCID: essential role of Jak3 in lymphoid development." Science 270.5237 (November 3, 1995): 797-800.
PMID
7481768
Source
pubmed
Published In
Science
Volume
270
Issue
5237
Publish Date
1995
Start Page
797
End Page
800

Correction of proliferative responses in purine nucleoside phosphorylase (PNP)-deficient T lymphocytes by retroviral-mediated PNP gene transfer and expression.

Purine nucleoside phosphorylase (PNP; EC 2.4.2.1) deficiency is associated with a fatal T cell immunodeficiency in children, a candidate condition for gene therapy by introduction of functional PNP sequences into either T lymphocytes or more primitive progenitor cells in the bone marrow. To test the effectiveness of PNP gene transfer in T lymphocytes, a retroviral vector (LmPSN-2) was designed and constructed to express the murine PNP cDNA under transcriptional regulation of the Moloney murine leukemia virus long terminal repeat. LmPSN-2 was first used to mediate gene transfer and expression of electrophoretically distinct murine PNP in normal (PNP-positive) human PBL. Peripheral blood leukocytes were then obtained from a PNP deficient patient and characterized phenotypically. Despite their paucity and general mitogenic unresponsiveness, T lymphocytes from this patient were successfully grown in culture by using anti-CD3 with rIL-2 and then transduced with LmPSN-2. Elevated PNP enzyme activity was observed in the transduced cell population. Mitogenic and allogeneic responses, normally depressed in PNP-deficient patients' cells, were partially corrected in the transduced cell population relative to nontransduced cells. These results suggest the possibility of effecting improved immunologic function in PNP-deficient T lymphoid cells by retroviral-mediated gene transfer as therapy for PNP deficiency.

Authors
Nelson, DM; Butters, KA; Markert, ML; Reinsmoen, NL; McIvor, RS
MLA Citation
Nelson, DM, Butters, KA, Markert, ML, Reinsmoen, NL, and McIvor, RS. "Correction of proliferative responses in purine nucleoside phosphorylase (PNP)-deficient T lymphocytes by retroviral-mediated PNP gene transfer and expression." J Immunol 154.6 (March 15, 1995): 3006-3014.
PMID
7876563
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
154
Issue
6
Publish Date
1995
Start Page
3006
End Page
3014

Molecular basis of adenosine deaminase deficiency.

Adenosine deaminase (ADA) deficiency is an autosomal recessive disorder resulting in immunodeficiency. Since the cDNA for ADA was cloned approximately 10 years ago, investigators have determined the molecular basis for disease in many patients with ADA deficiency. Mutations that have been identified include point mutations causing amino acid substitutions, premature stop codons, RNA splicing errors, and deletion mutations. Approximately one third of patients are homozygous for their mutation; in some of these cases the parents are known to be related. One mutation, Ala329-Val, is the most common, being present in 8 of the 21 ADA-deficient SCID patients whose mutations have been reported.

Authors
Markert, ML
MLA Citation
Markert, ML. "Molecular basis of adenosine deaminase deficiency." Immunodeficiency 5.2 (1994): 141-157. (Review)
PMID
8032366
Source
pubmed
Published In
Immunodeficiency
Volume
5
Issue
2
Publish Date
1994
Start Page
141
End Page
157

Haploidentical bone marrow stem cell transplantation in human severe combined immunodeficiency.

From May 1992 to March 1993, 50 infants with severe combined immunodeficiency (SCID) were given bone marrow transplants at Duke University Medical Center. None received chemotherapy for conditioning or for graft-versus-host disease (GVHD) prophylaxis. Forty-one received haploidentical parental marrow depleted of T cells by soybean lectin and sheep red blood cell resetting, and nine received HLA-identical marrow. Forty (80%) survived from 1 week to almost 11 years posttransplantation, including nine of nine (100%) HLA-identical marrow recipients and 31 of 41 haploidentical recipients. T-cell function was present within 2 weeks after transplantation of unfractionated HLA-identical marrow, but not until 3 to 4 months after T-cell-depleted haploidentical marrow stem cells. All 37 patients who are more than 4 months posttransplantation have good T-cell function, and all but one have 100% donor T cells. B-cell function developed slowly or not at all in some recipients of haploidentical marrow. Fourteen (four HLA-identical and 10 haploidentical recipients) have some donor B cells; 19 patients are receiving intravenous immune globulin (IVIG) therapy.

Authors
Buckley, RH; Schiff, SE; Schiff, RI; Roberts, JL; Markert, ML; Peters, W; Williams, LW; Ward, FE
MLA Citation
Buckley, RH, Schiff, SE, Schiff, RI, Roberts, JL, Markert, ML, Peters, W, Williams, LW, and Ward, FE. "Haploidentical bone marrow stem cell transplantation in human severe combined immunodeficiency." Semin Hematol 30.4 Suppl 4 (October 1993): 92-101. (Review)
PMID
7905667
Source
pubmed
Published In
Seminars in Hematology
Volume
30
Issue
4 Suppl 4
Publish Date
1993
Start Page
92
End Page
101

A homozygous 5 base-pair deletion in exon 10 of the adenosine deaminase (ADA) gene in a child with severe combined immunodeficiency and very low levels of ADA mRNA and protein.

Authors
Gossage, DL; Norby-Slycord, CJ; Hershfield, MS; Markert, ML
MLA Citation
Gossage, DL, Norby-Slycord, CJ, Hershfield, MS, and Markert, ML. "A homozygous 5 base-pair deletion in exon 10 of the adenosine deaminase (ADA) gene in a child with severe combined immunodeficiency and very low levels of ADA mRNA and protein." Hum Mol Genet 2.9 (September 1993): 1493-1494.
PMID
8242080
Source
pubmed
Published In
Human Molecular Genetics
Volume
2
Issue
9
Publish Date
1993
Start Page
1493
End Page
1494

A missense mutation in exon 4 of the human adenosine deaminase gene causes severe combined immunodeficiency.

Authors
Atasoy, U; Norby-Slycord, CJ; Markert, ML
MLA Citation
Atasoy, U, Norby-Slycord, CJ, and Markert, ML. "A missense mutation in exon 4 of the human adenosine deaminase gene causes severe combined immunodeficiency." Hum Mol Genet 2.8 (August 1993): 1307-1308.
PMID
7691348
Source
pubmed
Published In
Human Molecular Genetics
Volume
2
Issue
8
Publish Date
1993
Start Page
1307
End Page
1308

Haploidentical bone marrow stem cell transplantation in human severe combined immunodeficiency

From May 1992 to March 1993, 50 infants with severe combined immunodeficiency (SCID) were given bone marrow transplants at Duke University Medical Center. None received chemotherapy for conditioning or for graft- versus-host disease (GVHD) prophylaxis. Forty-one received haploidentical parental marrow depleted of T cells by soybean lectin and sheep red blood cell resetting, and nine received HLA-identical marrow. Forty (80%) survived from 1 week to almost 11 years postransplantation, including nine of nine (100%) HLA-identical marrow recipients and 31 of 41 haploidentical recipients. T-cell function was present within 2 weeks after transplantation of unfractionated HLA-identical marrow, but not until 3 to 4 months after T- cell-depleted haploidentical marrow stem cells. All 37 patients who are more than 4 months posttransplantation have good T-cell function, and all but one have 100% donor T cells. B-cell function developed slowly or not at all in some recipients of haploidentical marrow. Fourteen (four HLA-identical and 10 haploidentical recipients) have some donor B cells; 19 patients are receiving intravenous immune globulin (IVIG) therapy.

Authors
Buckley, RH; Schiff, SE; Schiff, RI; Roberts, JL; Markert, ML; Peters, W; Williams, LW; Ward, FE; Ballow, ; Boxer, ; Strober, ; McGhee, ; Schwartz, ; Nelson, ; Puck, ; Ochs, ; Parkman, ; Deeg,
MLA Citation
Buckley, RH, Schiff, SE, Schiff, RI, Roberts, JL, Markert, ML, Peters, W, Williams, LW, Ward, FE, Ballow, , Boxer, , Strober, , McGhee, , Schwartz, , Nelson, , Puck, , Ochs, , Parkman, , and Deeg, . "Haploidentical bone marrow stem cell transplantation in human severe combined immunodeficiency." Seminars in Hematology 30.4 SUPPL. 4 (1993): 92-104.
Source
scival
Published In
Seminars in Hematology
Volume
30
Issue
4 SUPPL. 4
Publish Date
1993
Start Page
92
End Page
104

Molecular analysis of mutations in a patient with purine nucleoside phosphorylase deficiency.

Purine nucleoside phosphorylase (PNP) deficiency is an inherited autosomal recessive disorder resulting in severe combined immunodeficiency. The purpose of this study was to determine the molecular defects responsible for PNP deficiency in one such patient. The patient's PNP cDNA was amplified by PCR and sequenced. Point mutations leading to amino acid substitutions were found in both alleles. One point mutation led to a Ser-to-Gly substitution at amino acid 51 and was common to both alleles. In addition, an Asp-to-Gly substitution at amino acid 128 and an Arg-to-Pro substitution at amino acid 234 were found in the maternal and paternal alleles, respectively. In order to prove that these mutations were responsible for the disease state, each of the three mutations was constructed separately by site-directed mutagenesis of the normal PNP cDNA, and each was transiently expressed in COS cells. Lysates from cells transfected with the allele carrying the substitution at amino acid 51 retained both function and immunoreactivity. Lysates from cells transfected with PNP alleles carrying a substitution at either amino acid 128 or amino acid 234 contained immunoreactive material but had no detectable human PNP activity. In summary, molecular analysis of this patient identified point mutations within the PNP gene which are responsible for the enzyme deficiency.

Authors
Aust, MR; Andrews, LG; Barrett, MJ; Norby-Slycord, CJ; Markert, ML
MLA Citation
Aust, MR, Andrews, LG, Barrett, MJ, Norby-Slycord, CJ, and Markert, ML. "Molecular analysis of mutations in a patient with purine nucleoside phosphorylase deficiency." Am J Hum Genet 51.4 (October 1992): 763-772.
PMID
1384322
Source
pubmed
Published In
The American Journal of Human Genetics
Volume
51
Issue
4
Publish Date
1992
Start Page
763
End Page
772

Exon skipping in purine nucleoside phosphorylase mRNA processing leading to severe immunodeficiency.

We report a defect in splicing of precursor messenger RNA (pre-mRNA) resulting from a naturally occurring mutation of the gene encoding purine nucleoside phosphorylase (PNP) in a patient with PNP-deficient severe combined immunodeficiency. This defects results from a G to T transversion at the terminal nucleotide of exon 2 within the 5' splice site of intron 2 and causes skipping of exon 2 during processing of PNP pre-mRNA. Translation of the misspliced mRNA results in a reading frameshift at the exon 1-exon 3 junction. The predicted polypeptide encoded by the aberrant mRNA is severely truncated, terminating at 31 amino acids. Only 4 residues at the NH2 terminus of the polypeptide correspond to PNP amino acids. Otherwise the translation product of the misspliced mRNA differs completely from PNP in amino acid sequence and has no PNP activity. The finding of exon skipping in PNP is the first report of a splicing defect resulting in PNP-deficient severe combined immunodeficiency. Analysis of the genomic context of the G-1 to T mutation of the 5' splice site lends support for the exon definition model of pre-mRNA splicing and contributes to the understanding of splice site selection.

Authors
Andrews, LG; Markert, ML
MLA Citation
Andrews, LG, and Markert, ML. "Exon skipping in purine nucleoside phosphorylase mRNA processing leading to severe immunodeficiency." J Biol Chem 267.11 (April 15, 1992): 7834-7838.
PMID
1560016
Source
pubmed
Published In
The Journal of biological chemistry
Volume
267
Issue
11
Publish Date
1992
Start Page
7834
End Page
7838

Molecular biology and allergy: Current status and future prospects

Authors
Markert, ML
MLA Citation
Markert, ML. "Molecular biology and allergy: Current status and future prospects." Pediatric Allergy and Immunology 3.2 (1992): 49-60.
Source
scival
Published In
Pediatric Allergy and Immunology
Volume
3
Issue
2
Publish Date
1992
Start Page
49
End Page
60

Purine nucleoside phosphorylase deficiency.

Purine nucleoside phosphorylase (PNP) deficiency is a rare inherited disease accounting for approximately 4% of patients with severe combined immunodeficiency. Thirty-three patients have been reported. PNP-deficient patients suffer from recurrent infections, usually beginning in the first year of life. Two thirds of patients have evidence of neurologic disorders. Findings range from spasticity to developmental delay, to mental retardation. One third of patients develop autoimmune disease. The most common manifestation of this is autoimmune hemolytic anemia. Idiopathic thrombocytopenic purpura and systemic lupus erythematosis have also been reported. Patients usually present with infections but approximately one fourth have come to medical care initially for neurological problems. In PNP deficiency, T- and B-cell immunity are affected. T-cell function may be profoundly deficient, may be normal at birth and then decrease with time, or may fluctuate repeatedly between low and normal. B-cell function can be normal but is deficient in approximately one third of patients. PNP protein is a trimer of approximately 90,000 daltons. It is found in most tissues of the body but is at highest levels in lymphoid tissues. This tissue distribution explains why the lymphoid system is predominantly affected in PNP deficiency. Many mechanisms have been proposed to explain the metabolic toxicity in PNP deficiency. The elevated dGTP found in PNP deficiency is thought to inhibit ribonucleotide reductase and, thus, impede cell division. Depressed GTP levels may correlate with neurologic dysfunction. The gene for PNP has been cloned; it is located on the long arm of chromosome 14. Studies of a mutant PNP gene isolated from one patient showed that a point mutation resulting in an amino acid substitution was responsible for PNP deficiency. PNP deficiency has a grave prognosis. No patient has reached the third decade of life. Twenty-nine of the 33 reported patients have died from their disease. Prenatal diagnosis is currently available. Many different therapies have been utilized for PNP deficiency including bone marrow transplantation, red cell transfusions, and supplementation of the diet with purines and pyrimidines. None of these therapies has been consistently successful. In light of the poor prognosis for PNP deficiency, bone marrow transplantation should be considered for all patients. In the future, improved forms of therapy such as gene therapy may become available.

Authors
Markert, ML
MLA Citation
Markert, ML. "Purine nucleoside phosphorylase deficiency." Immunodefic Rev 3.1 (1991): 45-81. (Review)
PMID
1931007
Source
pubmed
Published In
Immunodeficiency Reviews
Volume
3
Issue
1
Publish Date
1991
Start Page
45
End Page
81

A high proportion of ADA point mutations associated with a specific alanine-to-valine substitution.

In 15%-20% of children with severe combined immunodeficiency (SCID), the underlying defect is adenosine deaminase (ADA) deficiency. The overall goal of our research has been to identify the precise molecular defects in patients with ADA-deficient SCID. In this study, we focused on a patient whom we found to have normal sized ADA mRNA by Northern analysis and an intact ADA structural gene by Southern analysis. By cloning and sequencing this patient's ADA cDNA, we found a C-to-T point mutation in exon 11. This resulted in the amino acid substitution of a valine for an alanine at position 329 of the ADA protein. Sequence analysis revealed that this mutation created a new BalI restriction site. Using Southern analyses, we were able to directly screen individuals to determine the frequency of this mutation. By combining data on eight families followed at our institution with data on five other families reported in the literature, we established that five of 13 patients (seven of 22 alleles) with known or suspected point mutations have this defect. This mutation was found to be associated with three different ADA haplotypes. This argues against a founder effect and suggests that the mutation is very old. In summary, a conservative amino acid substitution is found in a high proportion of patients with ADA deficiency; this can easily be detected by Southern analysis.

Authors
Markert, ML; Norby-Slycord, C; Ward, FE
MLA Citation
Markert, ML, Norby-Slycord, C, and Ward, FE. "A high proportion of ADA point mutations associated with a specific alanine-to-valine substitution." Am J Hum Genet 45.3 (September 1989): 354-361.
PMID
2773932
Source
pubmed
Published In
The American Journal of Human Genetics
Volume
45
Issue
3
Publish Date
1989
Start Page
354
End Page
361

Adenosine deaminase (ADA) deficiency due to deletion of the ADA gene promoter and first exon by homologous recombination between two Alu elements.

In 15-20% of children with severe combined immunodeficiency (SCID), the underlying defect is adenosine deaminase (ADA) deficiency. The goal of this study was to determine the precise molecular defect in a patient with ADA-deficient SCID whom we previously have shown to have a total absence of ADA mRNA and a structural alteration of the ADA gene. By detailed Southern analysis, we now have determined that the structural alteration is a deletion of approximately 3.3 kb, which included exon 1 and the promoter region of the ADA gene. DNA sequence analysis demonstrates that the deletion created a novel, complete Alu repeat by homologous recombination between two existing Alu repeats that flanked the deletion. The 26-bp recombination joint in the Alu sequence includes the 10-bp "B" sequence homologous to the RNA polymerase III promoter. This is the first example of homologous recombination involving the B sequence in Alu repeats. Similar recombination events have been identified involving Alu repeats in which the recombination joint was located between the A and B sequences of the polymerase III split promoter. The nonrandom location of these events suggests that these segments may be hot spots for recombination.

Authors
Markert, ML; Hutton, JJ; Wiginton, DA; States, JC; Kaufman, RE
MLA Citation
Markert, ML, Hutton, JJ, Wiginton, DA, States, JC, and Kaufman, RE. "Adenosine deaminase (ADA) deficiency due to deletion of the ADA gene promoter and first exon by homologous recombination between two Alu elements." J Clin Invest 81.5 (May 1988): 1323-1327.
PMID
3366897
Source
pubmed
Published In
Journal of Clinical Investigation
Volume
81
Issue
5
Publish Date
1988
Start Page
1323
End Page
1327
DOI
10.1172/JCI113458

Adenosine deaminase and purine nucleoside phosphorylase deficiencies: evaluation of therapeutic interventions in eight patients.

The courses of six patients with adenosine deaminase (ADA) and two with purine nucleoside phosphorylase (PNP) deficiencies were evaluated before and after therapy. The heterogeneity of immunologic and clinical parameters was striking in each enzyme deficiency. In both PNP and ADA deficiency, some patients had very low immunoglobulin levels, while others had normal levels. T-cell function was always low in patients with ADA deficiency. In the two patients with PNP deficiency, contrary to the classical descriptions of this disorder, T-cell function fluctuated with time. Five ADA-deficient patients were treated with irradiated normal red-cell transfusions as a form of enzyme replacement and showed no lasting benefit. Three of the ADA-deficient patients and one of the PNP-deficient patients were given transplants of haploidentical parental bone marrow stem cells without pretransplant immunosuppression. In the PNP-deficient patient, chimerism has not been documented on enzymatic testing. One ADA-deficient patient has demonstrated long-term engraftment with good B- and T-cell function. Haploidentical bone marrow transplantation is currently the preferred therapy for enzyme-deficient patients with absent T-cell function who do not have an HLA-identical donor, as it may result in a lasting reconstitution of immune function. In those patients with unsatisfactory responses to transplantation, however, specific enzyme replacement or gene therapy may be considered in the future.

Authors
Markert, ML; Hershfield, MS; Schiff, RI; Buckley, RH
MLA Citation
Markert, ML, Hershfield, MS, Schiff, RI, and Buckley, RH. "Adenosine deaminase and purine nucleoside phosphorylase deficiencies: evaluation of therapeutic interventions in eight patients." J Clin Immunol 7.5 (September 1987): 389-399.
PMID
3116034
Source
pubmed
Published In
Journal of Clinical Immunology
Volume
7
Issue
5
Publish Date
1987
Start Page
389
End Page
399

Identification of a deletion in the adenosine deaminase gene in a child with severe combined immunodeficiency.

A patient with adenosine deaminase-deficient severe combined immunodeficiency is described whose defect is secondary to deletion of a portion of the ADA structural gene. In Southern analyses, DNA from this patient does not hybridize to a genomic probe that includes the 3' end of exon 1. This implies that both his parents are heterozygous for deletions of exon 1 sequences. Consistent with this finding, the patient has no detectable adenosine deaminase mRNA by Northern analysis. This is the first report of a deletion mutation as the cause of adenosine deaminase deficiency.

Authors
Markert, ML; Hershfield, MS; Wiginton, DA; States, JC; Ward, FE; Bigner, SH; Buckley, RH; Kaufman, RE; Hutton, JJ
MLA Citation
Markert, ML, Hershfield, MS, Wiginton, DA, States, JC, Ward, FE, Bigner, SH, Buckley, RH, Kaufman, RE, and Hutton, JJ. "Identification of a deletion in the adenosine deaminase gene in a child with severe combined immunodeficiency." J Immunol 138.10 (May 15, 1987): 3203-3206.
PMID
3571974
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
138
Issue
10
Publish Date
1987
Start Page
3203
End Page
3206

TREATMENT OF ADENOSINE-DEAMINASE DEFICIENCY WITH POLYETHYLENE-GLYCOL MODIFIED ADENOSINE-DEAMINASE (PEG-ADA)

Authors
HERSHFIELD, MS; BUCKLEY, RH; GREENBERG, ML; MELTON, AL; KOBAYASHI, RH; KOBAYASHI, AL; SCHIFF, R; KURTZBERG, J; MARKERT, ML; ABUCHOWSKI, A
MLA Citation
HERSHFIELD, MS, BUCKLEY, RH, GREENBERG, ML, MELTON, AL, KOBAYASHI, RH, KOBAYASHI, AL, SCHIFF, R, KURTZBERG, J, MARKERT, ML, and ABUCHOWSKI, A. "TREATMENT OF ADENOSINE-DEAMINASE DEFICIENCY WITH POLYETHYLENE-GLYCOL MODIFIED ADENOSINE-DEAMINASE (PEG-ADA)." April 1987.
Source
wos-lite
Published In
Clinical Research
Volume
35
Issue
3
Publish Date
1987
Start Page
A593
End Page
A593

Treatment of adenosine deaminase deficiency with polyethylene glycol-modified adenosine deaminase.

We treated two children who had adenosine deaminase deficiency and severe combined immunodeficiency disease by injecting bovine adenosine deaminase modified by conjugation with polyethylene glycol. The modified enzyme was rapidly absorbed after intramuscular injection and had a half-life in plasma of 48 to 72 hours. Weekly doses of approximately 15 U per kilogram of body weight maintained plasma adenosine deaminase activity at two to three times the level of erythrocyte adenosine deaminase activity in normal subjects. The principal biochemical consequences of adenosine deaminase deficiency were almost completely reversed. In erythrocytes, adenosine nucleotides increased and deoxyadenosine nucleotides decreased to less than 0.5 percent of total adenine nucleotides. The activity of S-adenosylhomocysteine hydrolase, which is inactivated by deoxyadenosine, increased to normal in red cells and nucleated marrow cells. Neither toxic effects nor hypersensitivity reactions were observed. In vitro tests of the cellular immune function of each patient showed marked improvement, along with an increase in circulating T lymphocytes. Clinical improvement was indicated by absence of infection and resumption of weight gain. We conclude that from the standpoints of efficacy, convenience, and safety, polyethylene glycol-modified adenosine deaminase is preferable to red-cell transfusion as a treatment for adenosine deaminase deficiency. Patients with other inherited metabolic diseases in which accumulated metabolites equilibrate with plasma could benefit from treatment with the appropriate polyethylene glycol-modified enzyme.

Authors
Hershfield, MS; Buckley, RH; Greenberg, ML; Melton, AL; Schiff, R; Hatem, C; Kurtzberg, J; Markert, ML; Kobayashi, RH; Kobayashi, AL
MLA Citation
Hershfield, MS, Buckley, RH, Greenberg, ML, Melton, AL, Schiff, R, Hatem, C, Kurtzberg, J, Markert, ML, Kobayashi, RH, and Kobayashi, AL. "Treatment of adenosine deaminase deficiency with polyethylene glycol-modified adenosine deaminase." N Engl J Med 316.10 (March 5, 1987): 589-596.
PMID
3807953
Source
pubmed
Published In
The New England journal of medicine
Volume
316
Issue
10
Publish Date
1987
Start Page
589
End Page
596
DOI
10.1056/NEJM198703053161005

Natural-killer-cell function and bone marrow transplantation.

Authors
Markert, ML; Buckley, RH
MLA Citation
Markert, ML, and Buckley, RH. "Natural-killer-cell function and bone marrow transplantation." N Engl J Med 315.22 (November 27, 1986): 1418-1419. (Letter)
PMID
3534572
Source
pubmed
Published In
The New England journal of medicine
Volume
315
Issue
22
Publish Date
1986
Start Page
1418
End Page
1419
DOI
10.1056/NEJM198611273152216

Development of immunity in human severe primary T cell deficiency following haploidentical bone marrow stem cell transplantation.

Recent advances in the prevention of graft-vs-host disease (GVHD) have allowed the use of haploidentical bone marrow cells for correction of lethal genetic defects of the immune system. Sequential analyses of blood lymphocyte phenotypes and functions were done before and after transplantation of haploidentical marrow stem cells into 17 infants with severe primary T cell deficiencies. The marrow was depleted of post-thymic T cells and most other mature marrow cells by soy lectin agglutination and sheep erythrocyte rosetting. The studies were performed to define the time course and extent of appearance of immune function, and to identify factors leading to resistance to engraftment. No pretransplant immunosuppression was used. T cell function was detected between 34 and 287 days after transplantation, but a sharp rise usually occurred between 84 and 115 days, and normal function was reached between 113 and 210 days. Fifteen of the patients are alive from 6 to 41 mo post-transplantation, 12 have improved or have normal T lymphocyte function, and nine have proven T cell chimerism. Increased immunoglobulins of several isotypes have been noted in 11 patients and specific antibodies in seven patients, although B cell chimerism has been detected in only one patient. B cell function required 2 to 2.5 yr for normalization. No GVHD occurred in 14 patients, and the other three had only transient mild skin rashes. Two patients died of viral infections. Failure to engraft was correlated with some pre-transplant lymphocyte responses to mitogens and allogeneic cells (three cases), but not with the presence of pre-transplant natural killer cell function (five cases) nor with the presence of purine salvage pathway enzyme deficiencies (four cases). The latter, however, was associated with poor lymphoid function in two patients. These studies indicate that the thymic microenvironment of most infants with severe combined immunodeficiency disease is capable of differentiating donor stem cells to mature and functioning T lymphocytes which can cooperate with apparently normal host B cells for antibody production.

Authors
Buckley, RH; Schiff, SE; Sampson, HA; Schiff, RI; Markert, ML; Knutsen, AP; Hershfield, MS; Huang, AT; Mickey, GH; Ward, FE
MLA Citation
Buckley, RH, Schiff, SE, Sampson, HA, Schiff, RI, Markert, ML, Knutsen, AP, Hershfield, MS, Huang, AT, Mickey, GH, and Ward, FE. "Development of immunity in human severe primary T cell deficiency following haploidentical bone marrow stem cell transplantation." J Immunol 136.7 (April 1, 1986): 2398-2407.
PMID
2869085
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
136
Issue
7
Publish Date
1986
Start Page
2398
End Page
2407

Human B cell alloantigens: expression of MB and MT determinants.

Four HLA-DR-homozygous cell lines were examined in detail to compare alpha and beta subunits precipitated by chimpanzee antisera and alloantisera directed against HLA-DR, MB, or MT determinants. For the HLA-DR5-homozygous cell line Sweig, chimpanzee antisera directed against MT2 and against the HLA-DR5 specificity yielded identical patterns. Similar findings for MT and MB vs HLA-DR were obtained for the cell lines LG14 (HLA-DR3-homozygous), PBur (HLA-DR7-homozygous), and Eld (HLA-DRw6-homozygous). Therefore, on some cell lines MT and/or MB specificities appear to be detectable only on the same or electrophoretically identical alpha-beta complexes as HLA-DR determinants.

Authors
Markert, ML; Cresswell, P
MLA Citation
Markert, ML, and Cresswell, P. "Human B cell alloantigens: expression of MB and MT determinants." J Immunol 128.5 (May 1982): 2004-2008.
PMID
6174619
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
128
Issue
5
Publish Date
1982
Start Page
2004
End Page
2008

Human B cell alloantigens; alpha subunit variability.

Human B cell alloantigens were studied by two-dimensional gel electrophoresis. The heavy (alpha) and light (beta) chains precipitated by xenoantisera were compared on B lymphoblastoid cell lines of different HLA-DR types. In accordance with the results of other investigators, we observed considerable electrophoretic variability of beta subunits. Significant electrophoretic differences between alpha subunits isolated from different cells were also observed through the use of a pH 5 to 7 gradient in the isoelectric focusing dimension. HLA-DR allelic products from a heterozygous line exhibited differences in beta subunit electrophoretic mobility but apparent identity of alpha subunits.

Authors
Markert, ML; Cresswell, P
MLA Citation
Markert, ML, and Cresswell, P. "Human B cell alloantigens; alpha subunit variability." J Immunol 128.5 (May 1982): 1999-2003.
PMID
6977579
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
128
Issue
5
Publish Date
1982
Start Page
1999
End Page
2003

Polymorphism of human B-cell alloantigens: evidence for three loci within the HLA system.

The B-cell alloantigens of an HLA-DR3-homozygous, MB2-homozygous, MT2-positive lymphoblastoid cell line were studied by two-dimensional gel electrophoresis. Analysis of gel patterns suggested assignment of the HLA-DR3 determinant to the larger (35,000-dalton) subunit (alpha) of the B-cell alloantigen. MB2 was found to be either a determinant on the small (27,000 dalton) subunit (beta) or a crossreactive determinant(s) on the HLA-DR3 alpha subunit and an additional alpha subunit. MT2 was found to be a determinant on Ia antigen-like molecules distinct from those carrying the MB and HLA-DR determinants. The results are consistent with the existence in the major histocompatibility complex of at least three loci encoding B-cell alloantigens.

Authors
Markert, ML; Cresswell, P
MLA Citation
Markert, ML, and Cresswell, P. "Polymorphism of human B-cell alloantigens: evidence for three loci within the HLA system." Proc Natl Acad Sci U S A 77.10 (October 1980): 6101-6104.
PMID
6160591
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
77
Issue
10
Publish Date
1980
Start Page
6101
End Page
6104

Polymorphism of human B-cell alloantigens: Evidence for three loci within the HLA system

The B-cell alloantigens of an HLA-DR3-homozygous, MB2-homozygous, MT2-positive lymphoblastoid cell line were studied by two-dimensional gel electrophoresis. Analysis of gel patterns suggested assignment of the HLA-DR3 determinant to the larger (35,000-dalton) subunit (α) of the B-cell alloantigen. MB2 was found to be either a determinant on the small (27,000 dalton) subunit (β) or a crossreactive determinant(s) on the HLA-DR3 α subunit and an additional α subunit, MT2 was found to be a determinant on Ia antigen-like molecules distinct from those carrying the MB and HLA-DR determinants. The results are consistent with the existence in the major histocompatibility complex of at least three loci encoding B-cell alloantigens.

Authors
Markert, ML; Cresswell, P
MLA Citation
Markert, ML, and Cresswell, P. "Polymorphism of human B-cell alloantigens: Evidence for three loci within the HLA system." Proceedings of the National Academy of Sciences of the United States of America 77.10 II (1980): 6101-6104.
Source
scival
Published In
Proceedings of the National Academy of Sciences of the United States of America
Volume
77
Issue
10 II
Publish Date
1980
Start Page
6101
End Page
6104
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Research Areas:

  • Immunologic Deficiency Syndromes
  • Thymus Gland
  • Transplantation
  • Transplantation Tolerance