Bio

Clinical Focus


  • Immunology and Rheumatology
  • Allergy and Immunology

Academic Appointments


Administrative Appointments


  • Council, Midwinter Conference of Immunologists (1981 - 1986)
  • Council, American Society for Clinical Investigation (1984 - 1987)
  • Associate Editor, Annual Review of Immunology (1981 - 2005)
  • Division Chief, Division of Immunology and Rheumatology (1997 - 2012)
  • Director, Center for Clinical Immunology at Stanford (CCIS) (1993 - Present)
  • President, Clinical Immunology Society (2000 - 2001)
  • Associate Director, ITI Institute Stanford (2008 - Present)
  • President, Federation of Clinical Immunology Societies (FOCIS) (2002 - 2005)
  • Division Chief, Division of Immunology and Rheumatology, Stanford University Medical Center (1997 - Present)

Honors & Awards


  • Division Teaching Award, Stanford University School of Medicine, Department of Medicine (2011)
  • Founder's Award, Federation of Clinical immunology Societies (2010)
  • Master, American College of Rheumatology (2007)
  • President's Award, Clinical Immunology Society (2006)
  • Alumni Achievement Award, Washington University Medical School (1999)
  • Naomi M. Kanof Award for Distinguished Achievement in Clinical Investigation, Society for Investigative Dermatology (1997)

Professional Education


  • Residency:Mary Hitchcock Mem Hosp (1971) NH
  • Medical Education:Washington University School of Medicine (1969) MO
  • Fellowship:Stanford University School of Medicine (1973) CA
  • Fellowship:National Institutes of Health (1975) MD
  • Internship:Mary Hitchcock Mem Hosp (1970) NH
  • B.A., Univ. Kentucky, Lexington, Pre-Med (1964)
  • M.D., Washington Univ., St. Louis, Medicine (1969)

Research & Scholarship

Current Research and Scholarly Interests


My laboratory of molecular and cellular immunology is interested in mechanisms of T cell anergy and the pathophysiology and immunotherapy of preclinical animal models of autoimmune disease.
I. T Cell Anergy: We have identified a ubiquitin E3 ligase (GRAIL) that seems to be central to the control of T cell anergy. We developed a novel prokaryotic system to screen for E3 ligase substrates and identified RhoGDI as an E3 substrate of GRAIL. Using two-hybrid technology to identify proteins that bound the lumenal or extracellular domain of GRAIL identified CD151, a member of the tetraspanin family of membrane proteins and GRAIL over-expression promoted polyubiquitination of all tested tetraspanins. These findings identify for the first time an E3 ligase with a substrate-binding domain spatially restricted by a membrane from its ubiquitination machinery. Recent studies focus on understanding GRAIL mediated CD4 T cell unresponsiveness. Two deubiquiting enzyes, USP8 and OTUB1, play contrasting roles in maintaining GRAIL stability
II. Gene Therapy: We have demonstrated that the local delivery of anti-inflammatory proteins via adoptive cellular gene therapy using syngeneic dendritic cells (DCs) transduced to express immunoregulatory proteins, in three murine models of autoimmunity (RA, MS and T1D), provide therapeutic effect both in the prevention of disease onset and in therapy of established disease.
III. Gene expression studies in autoimmunity: The major emphasis placed on disease associated genetic mutations or polymorphisms to understand the genetics of T1D has failed to advance either understanding of T1D pathogenesis or to identify therapeutic targets. Recent studies from my lab have demonstrated that tissue- and disease-specific changes in mRNA expression, rather than DNA variants, may underlie the progression of T1D. By combining the expertise of the lab in T1D research with established preclinical models and patient samples/tissues from the Network for Pancreatic Organ Donors with Diabetes, nPOD (http://www.jdrfnpod.org/), my lab has both demonstrated a potential defect in peripheral tolerance in NOD mice that has homologies in T1D patients and have demonstrated that appropriate immunotherapy may overcome this defect.
iv. Mechanism of action of biologic therapy: Our inability to identify subgroups of RA patients who will or will not respond to a biologic therapy is frustrating to clinicians and patients and a major burden to the health care economy. My lab is currently employing new multiplexed technologies in proteomic and genomic analysis of the response of patients’ PBCs to activation with one of the cytokines (TNF) thought to be a major pathophysiological component of RA. By analyzing the response of RA patient PBCs to TNF [using gene expression of PBCs by microarray technology and algorithms recently developed at Stanford that allow total PBC gene expression to be deconvoluted to know which cell subset expressed which subset of genes, and the new cytometry time of flight (CyTOF) proteomic technology developed in Garry Nolan’s lab at Stanford], we hope to dissect the individual patients’ PBC gene expression or protein modification pattern in response to TNF activation to identify responders from non-responders to TNF inhibitor therapies; commercially available and FDA approved therapeutics

Teaching

2013-14 Courses


Postdoctoral Advisees


Publications

Journal Articles


  • Reduced DEAF1 function during type 1 diabetes inhibits translation in lymph node stromal cells by suppressing Eif4g3 JOURNAL OF MOLECULAR CELL BIOLOGY Yip, L., Creusot, R. J., Pager, C. T., Sarnow, P., Fathman, C. G. 2013; 5 (2): 99-110

    Abstract

    The transcriptional regulator deformed epidermal autoregulatory factor 1 (DEAF1) has been suggested to play a role in maintaining peripheral tolerance by controlling the transcription of peripheral tissue antigen genes in lymph node stromal cells (LNSCs). Here, we demonstrate that DEAF1 also regulates the translation of genes in LNSCs by controlling the transcription of the poorly characterized eukaryotic translation initiation factor 4 gamma 3 (Eif4g3) that encodes eIF4GII. Eif4g3 gene expression was reduced in the pancreatic lymph nodes of Deaf1-KO mice, non-obese diabetic mice, and type 1 diabetes patients, where functional Deaf1 is absent or diminished. Silencing of Deaf1 reduced Eif4g3 expression, but increased the expression of Caspase 3, a serine protease that degrades eIF4GII. Polysome profiling showed that reduced Eif4g3 expression in LNSCs resulted in the diminished translation of various genes, including Anpep, the gene for aminopeptidase N, an enzyme involved in fine-tuning antigen presentation on major histocompatibility complex (MHC) class II. Together these findings suggest that reduced DEAF1 function, and subsequent loss of Eif4g3 transcription may affect peripheral tissue antigen (PTA) expression in LNSCs and contribute to the pathology of T1D.

    View details for DOI 10.1093/jmcb/mjs052

    View details for Web of Science ID 000316701200004

    View details for PubMedID 22923498

  • New tools for classification and monitoring of autoimmune diseases NATURE REVIEWS RHEUMATOLOGY Maecker, H. T., Lindstrom, T. M., Robinson, W. H., Utz, P. J., Hale, M., Boyd, S. D., Shen-Orr, S. S., Fathman, C. G. 2012; 8 (6): 317-328

    Abstract

    Rheumatologists see patients with a range of autoimmune diseases. Phenotyping these diseases for diagnosis, prognosis and selection of therapies is an ever increasing problem. Advances in multiplexed assay technology at the gene, protein, and cellular level have enabled the identification of 'actionable biomarkers'; that is, biological metrics that can inform clinical practice. Not only will such biomarkers yield insight into the development, remission, and exacerbation of a disease, they will undoubtedly improve diagnostic sensitivity and accuracy of classification, and ultimately guide treatment. This Review provides an introduction to these powerful technologies that could promote the identification of actionable biomarkers, including mass cytometry, protein arrays, and immunoglobulin and T-cell receptor high-throughput sequencing. In our opinion, these technologies should become part of routine clinical practice for the management of autoimmune diseases. The use of analytical tools to deconvolve the data obtained from use of these technologies is also presented here. These analyses are revealing a more comprehensive and interconnected view of the immune system than ever before and should have an important role in directing future treatment approaches for autoimmune diseases.

    View details for DOI 10.1038/nrrheum.2012.66

    View details for Web of Science ID 000304719600005

    View details for PubMedID 22647780

  • Exploiting a natural conformational switch to engineer an interleukin-2 'superkine' NATURE Levin, A. M., Bates, D. L., Ring, A. M., Krieg, C., Lin, J. T., Su, L., Moraga, I., Raeber, M. E., Bowman, G. R., Novick, P., Pande, V. S., Fathman, C. G., Boyman, O., Garcia, K. C. 2012; 484 (7395): 529-U159

    Abstract

    The immunostimulatory cytokine interleukin-2 (IL-2) is a growth factor for a wide range of leukocytes, including T cells and natural killer (NK) cells. Considerable effort has been invested in using IL-2 as a therapeutic agent for a variety of immune disorders ranging from AIDS to cancer. However, adverse effects have limited its use in the clinic. On activated T cells, IL-2 signals through a quaternary 'high affinity' receptor complex consisting of IL-2, IL-2R? (termed CD25), IL-2R? and IL-2R?. Naive T cells express only a low density of IL-2R? and IL-2R?, and are therefore relatively insensitive to IL-2, but acquire sensitivity after CD25 expression, which captures the cytokine and presents it to IL-2R? and IL-2R?. Here, using in vitro evolution, we eliminated the functional requirement of IL-2 for CD25 expression by engineering an IL-2 'superkine' (also called super-2) with increased binding affinity for IL-2R?. Crystal structures of the IL-2 superkine in free and receptor-bound forms showed that the evolved mutations are principally in the core of the cytokine, and molecular dynamics simulations indicated that the evolved mutations stabilized IL-2, reducing the flexibility of a helix in the IL-2R? binding site, into an optimized receptor-binding conformation resembling that when bound to CD25. The evolved mutations in the IL-2 superkine recapitulated the functional role of CD25 by eliciting potent phosphorylation of STAT5 and vigorous proliferation of T cells irrespective of CD25 expression. Compared to IL-2, the IL-2 superkine induced superior expansion of cytotoxic T cells, leading to improved antitumour responses in vivo, and elicited proportionally less expansion of T regulatory cells and reduced pulmonary oedema. Collectively, we show that in vitro evolution has mimicked the functional role of CD25 in enhancing IL-2 potency and regulating target cell specificity, which has implications for immunotherapy.

    View details for DOI 10.1038/nature10975

    View details for Web of Science ID 000303200400054

    View details for PubMedID 22446627

  • Therapeutic Effects of Systemic Administration of Chaperone alpha B-Crystallin Associated with Binding Proinflammatory Plasma Proteins JOURNAL OF BIOLOGICAL CHEMISTRY Rothbard, J. B., Kurnellas, M. P., Brownell, S., Adams, C. M., Su, L., Axtell, R. C., Chen, R., Fathman, C. G., Robinson, W. H., Steinman, L. 2012; 287 (13): 9708-9721

    Abstract

    The therapeutic benefit of the small heat shock protein ?B-crystallin (HspB5) in animal models of multiple sclerosis and ischemia is proposed to arise from its increased capacity to bind proinflammatory proteins at the elevated temperatures within inflammatory foci. By mass spectral analysis, a common set of ?70 ligands was precipitated by HspB5 from plasma from patients with multiple sclerosis, rheumatoid arthritis, and amyloidosis and mice with experimental allergic encephalomyelitis. These proteins were distinguished from other precipitated molecules because they were enriched in the precipitate as compared with their plasma concentrations, and they exhibited temperature-dependent binding. More than half of these ligands were acute phase proteins or members of the complement or coagulation cascades. Consistent with this proposal, plasma levels of HspB5 were increased in patients with multiple sclerosis as compared with normal individuals. The combination of the thermal sensitivity of the HspB5 combined with the high local concentration of these ligands at the site of inflammation is proposed to explain the paradox of how a protein believed to exhibit nonspecific binding can bind with some relative apparent selectivity to proinflammatory proteins and thereby modulate inflammation.

    View details for DOI 10.1074/jbc.M111.337691

    View details for Web of Science ID 000302167200005

    View details for PubMedID 22308023

  • Differential mTOR and ERK pathway utilization by effector CD4 T cells suggests combinatorial drug therapy of arthritis CLINICAL IMMUNOLOGY Lin, J. T., Stein, E. A., Wong, M. T., Kalpathy, K. J., Su, L. L., Utz, P. J., Robinson, W. H., Fathnnan, C. G. 2012; 142 (2): 127-138

    Abstract

    The signaling pathways utilized by naïve and experienced effector CD4 T cells during activation and proliferation were evaluated. While inhibition of either mTOR or MAPK alone was able to inhibit naïve T cell proliferation, both mTOR and MAPK (ERK) pathway inhibition was required to efficiently block experienced, effector CD4 T cell proliferation. This was demonstrated both in vitro, and in vivo by treating mice with collagen-induced arthritis using mTOR and/or ERK inhibitors. The combination of mTOR and ERK inhibition prevented or treated disease more efficiently than either agent alone. These data illustrate the different requirements of naïve and experienced effector CD4 T cells in the use of the mTOR and MAPK pathways in proliferation, and suggest that therapies targeting both the mTOR and MAPK pathways may be more effective than targeting either pathway alone in the treatment of CD4 T cell-mediated autoimmunity.

    View details for DOI 10.1016/j.clim.2011.09.008

    View details for Web of Science ID 000300388200007

    View details for PubMedID 22075384

  • GRAIL: a unique mediator of CD4 T-lymphocyte unresponsiveness FEBS JOURNAL Whiting, C. C., Su, L. L., Lin, J. T., Fathman, C. G. 2011; 278 (1): 47-58

    Abstract

    GRAIL (gene related to anergy in lymphocytes, also known as RNF128), an ubiquitin-protein ligase (E3), utilizes a unique single transmembrane protein with a split-function motif, and is an important gatekeeper of T-cell unresponsiveness. Although it may play a role in other CD4 T-cell functions including activation, survival and differentiation, GRAIL is most well characterized as a negative regulator of T-cell receptor responsiveness and cytokine production. Here, we review the recent literature on this remarkable E3 in the regulation of human and mouse CD4 T-cell unresponsiveness.

    View details for DOI 10.1111/j.1742-4658.2010.07922.x

    View details for Web of Science ID 000285249100006

    View details for PubMedID 21078124

  • A Short Pulse of IL-4 Delivered by DCs Electroporated With Modified mRNA Can Both Prevent and Treat Autoimmune Diabetes in NOD Mice MOLECULAR THERAPY Creusot, R. J., Chang, P., Healey, D. G., Tcherepanova, I. Y., Nicolette, C. A., Fathman, C. G. 2010; 18 (12): 2112-2120

    Abstract

    Bone marrow-derived dendritic cells (DCs) are cells of the immune system that have been used as a tool to boost, modulate, or dampen immune responses. In the context of autoimmunity, DCs can be modified to express immunoregulatory products encoded by transgenes, and used therapeutically in adoptive cellular therapy. DCs that were lentivirally transduced (lt) to express interleukin 4 (IL-4) can significantly delay or prevent the onset of autoimmune diabetes in nonobese diabetic (NOD) mice. However, modifying cells using viral vectors carries the dual risk of oncogenicity or immunogenicity. This study demonstrates that NOD DCs, electroporated with "translationally enhanced" IL-4 mRNA (eDC/IL-4), can be equally efficient therapeutically, despite the reduced amount and shorter duration of IL-4 secretion. Moreover, a single injection of eDC/IL-4 in NOD mice shortly after the onset of hyperglycemia was able to maintain stable glycemia for up to several months in a significant fraction of treated mice. Treatment with eDC/IL-4 boosted regulatory T (Tregs) cell functions and modulated T helper responses to reduce pathogenicity. Thus, treatment with DCs, electroporated with modified IL-4 mRNA to express IL-4 for up to 24 hours, constitutes a viable cellular therapy approach for the regulation of autoimmune diabetes, as a preferred alternative to the use of viral vectors.

    View details for DOI 10.1038/mt.2010.146

    View details for Web of Science ID 000284828000012

    View details for PubMedID 20628358

  • A model for harmonizing flow cytometry in clinical trials. Nature immunology Maecker, H. T., McCoy, J. P., Amos, M., Elliott, J., Gaigalas, A., Wang, L., Aranda, R., Banchereau, J., Boshoff, C., Braun, J., Korin, Y., Reed, E., Cho, J., Hafler, D., Davis, M., Fathman, C. G., Robinson, W., Denny, T., Weinhold, K., Desai, B., Diamond, B., Gregersen, P., Di Meglio, P., Nestle, F. O., Peakman, M., Villanova, F., Ferbas, J., Field, E., Kantor, A., Kawabata, T., Komocsar, W., Lotze, M., Nepom, J., Ochs, H., O'Lone, R., Phippard, D., Plevy, S., Rich, S., Roederer, M., Rotrosen, D., Yeh, J. 2010; 11 (11): 975-978

    Abstract

    Complexities in sample handling, instrument setup and data analysis are barriers to the effective use of flow cytometry to monitor immunological parameters in clinical trials. The novel use of a central laboratory may help mitigate these issues.

    View details for DOI 10.1038/ni1110-975

    View details for PubMedID 20959798

  • New technologies for autoimmune disease monitoring CURRENT OPINION IN ENDOCRINOLOGY DIABETES AND OBESITY Maecker, H. T., Nolan, G. P., Fathman, C. G. 2010; 17 (4): 322-328

    Abstract

    This article will review new technologies used to characterize the immune phenotype of cells and serum for potential use in studies of autoimmunity.One area of recent development in studies of immune phenotyping is the contrast between cells of the immune system at rest and following activation. This simply involves comparing these cells at rest and following ligand-induced activation and measuring signaling system activation (phosphoepitope identification) or intracellular cytokine production or activation-induced gene expression. Preliminary data using these techniques have begun to identify signatures of disease (biomarkers) that are only seen when using these activation-induced assays. One of the most exciting new technologies, cytometry by time-of-flight mass spectrometry, couples a flow cytometer to a mass spectrometer, allowing many more parameters to be analyzed per cell, and without spillover between assay reagents, compared to conventional optical flow cytometry (heavy ions, mass, replaces fluorophore readout). Another new technology to analyze soluble proteins, bead-based immunoassays, can simultaneously measure up to 75 soluble analytes in a multiplexed array. Other technologies provide similar innovations in terms of analytical content, throughput, and miniaturization.We believe that new cellular genomic and protein-based technologies can provide key insights into autoimmune disease pathogenesis, progression, and therapy, and that these assays need to be applied in a systematic way to samples from patients with autoimmune diseases.

    View details for DOI 10.1097/MED.0b013e32833ada91

    View details for Web of Science ID 000285063800003

    View details for PubMedID 20531181

  • Deaf1 isoforms control the expression of genes encoding peripheral tissue antigens in the pancreatic lymph nodes during type 1 diabetes NATURE IMMUNOLOGY Yip, L., Su, L., Sheng, D., Chang, P., Atkinson, M., Czesak, M., Albert, P. R., Collier, A., Turley, S. J., Fathman, C. G., Creusot, R. J. 2009; 10 (9): 1026-U107

    Abstract

    Type 1 diabetes may result from a breakdown in peripheral tolerance that is partially controlled by the expression of peripheral tissue antigens (PTAs) in lymph nodes. Here we show that the transcriptional regulator Deaf1 controls the expression of genes encoding PTAs in the pancreatic lymph nodes (PLNs). The expression of canonical Deaf1 was lower, whereas that of an alternatively spliced variant was higher, during the onset of destructive insulitis in the PLNs of nonobese diabetic (NOD) mice. We identified an equivalent variant Deaf1 isoform in the PLNs of patients with type 1 diabetes. Both the NOD mouse and human Deaf1 variant isoforms suppressed PTA expression by inhibiting the transcriptional activity of canonical Deaf1. Lower PTA expression resulting from the alternative splicing of DEAF1 may contribute to the pathogenesis of type 1 diabetes.

    View details for DOI 10.1038/ni.1773

    View details for Web of Science ID 000269120900017

    View details for PubMedID 19668219

  • The Transmembrane E3 Ligase GRAIL Ubiquitinates and Degrades CD83 on CD4 T Cells JOURNAL OF IMMUNOLOGY Su, L. L., Iwai, H., Lin, J. T., Fathman, C. G. 2009; 183 (1): 438-444

    Abstract

    Ubiquitination of eukaryotic proteins regulates a broad range of cellular processes, including T cell activation and tolerance. We have previously demonstrated that GRAIL (gene related to anergy in lymphocytes), a transmembrane RING finger ubiquitin E3 ligase, initially described as induced during the induction of CD4 T cell anergy, is also expressed in resting CD4 T cells. In this study, we show that GRAIL can down-modulate the expression of CD83 (previously described as a cell surface marker for mature dendritic cells) on CD4 T cells. GRAIL-mediated down-modulation of CD83 is dependent on an intact GRAIL extracellular protease-associated domain and an enzymatically active cytosolic RING domain, and proceeds via the ubiquitin-dependent 26S proteosome pathway. Ubiquitin modification of lysine residues K168 and K183, but not K192, in the cytoplasmic domain of CD83 was shown to be necessary for GRAIL-mediated degradation of CD83. Reduced CD83 surface expression levels were seen both on anergized CD4 T cells and following GRAIL expression by retroviral transduction, whereas GRAIL knock-down by RNA interference in CD4 T cells resulted in elevated CD83 levels. Furthermore, CD83 expression on CD4 T cells contributes to T cell activation as a costimulatory molecule. This study supports the novel mechanism of ubiquitination by GRAIL, identifies CD83 as a substrate of GRAIL, and ascribes a role for CD83 in CD4 T cell activation.

    View details for DOI 10.4049/jimmunol.0900204

    View details for Web of Science ID 000275119400048

    View details for PubMedID 19542455

  • Lymphoid tissue-specific homing of bone marrow-derived dendritic cells BLOOD Creusot, R. J., Yaghoubi, S. S., Chang, P., Chia, J., Contag, C. H., Gambhir, S. S., Fathman, C. G. 2009; 113 (26): 6638-6647

    Abstract

    Because of their potent immunoregulatory capacity, dendritic cells (DCs) have been exploited as therapeutic tools to boost immune responses against tumors or pathogens, or dampen autoimmune or allergic responses. Murine bone marrow-derived DCs (BM-DCs) are the closest known equivalent of the blood monocyte-derived DCs that have been used for human therapy. Current imaging methods have proven unable to properly address the migration of injected DCs to small and deep tissues in mice and humans. This study presents the first extensive analysis of BM-DC homing to lymph nodes (and other selected tissues) after intravenous and intraperitoneal inoculation. After intravenous delivery, DCs accumulated in the spleen, and preferentially in the pancreatic and lung-draining lymph nodes. In contrast, DCs injected intraperitoneally were found predominantly in peritoneal lymph nodes (pancreatic in particular), and in omentum-associated lymphoid tissue. This uneven distribution of BM-DCs, independent of the mouse strain and also observed within pancreatic lymph nodes, resulted in the uneven induction of immune response in different lymphoid tissues. These data have important implications for the design of systemic cellular therapy with DCs, and in particular underlie a previously unsuspected potential for specific treatment of diseases such as autoimmune diabetes and pancreatic cancer.

    View details for DOI 10.1182/blood-2009-02-204321

    View details for Web of Science ID 000267789600024

    View details for PubMedID 19363220

  • Naive CD4 T Cell Proliferation Is Controlled by Mammalian Target of Rapainaycin Regulation of GRAIL Expression JOURNAL OF IMMUNOLOGY Lin, J. T., Lineberry, N. B., Kattah, M. G., Su, L. L., Utz, P. J., Fathman, C. G., Wu, L. 2009; 182 (10): 5919-5928

    Abstract

    In this study, we demonstrate that the E3 ubiquitin ligase gene related to anergy in lymphocytes (GRAIL) is expressed in quiescent naive mouse and human CD4 T cells and has a functional role in inhibiting naive T cell proliferation. Following TCR engagement, CD28 costimulation results in the expression of IL-2 whose signaling through its receptor activates the Akt-mammalian target of rapamycin (mTOR) pathway. Activation of mTOR allows selective mRNA translation, including the epistatic regulator of GRAIL, Otubain-1 (Otub1), whose expression results in the degradation of GRAIL and allows T cell proliferation. The activation of mTOR appears to be the critical component of IL-2R signaling regulating GRAIL expression. CTLA4-Ig treatment blocks CD28 costimulation and resultant IL-2 expression, whereas rapamycin and anti-IL-2 treatment block mTOR activation downstream of IL-2R signaling. Thus, all three of these biotherapeutics inhibit mTOR-dependent translation of mRNA transcripts, resulting in blockade of Otub1 expression, maintenance of GRAIL, and inhibition of CD4 T cell proliferation. These observations provide a mechanistic pathway sequentially linking CD28 costimulation, IL-2R signaling, and mTOR activation as important requirements for naive CD4 T cell proliferation through the regulation of Otub1 and GRAIL expression. Our findings also extend the role of GRAIL beyond anergy induction and maintenance, suggesting that endogenous GRAIL regulates general cell cycle and proliferation of primary naive CD4 T cells.

    View details for DOI 10.4049/jimmunol.0803986

    View details for Web of Science ID 000265899800008

    View details for PubMedID 19414743

  • Tissue- and age-specific changes in gene expression during disease induction and progression in NOD mice CLINICAL IMMUNOLOGY Kodama, K., Butte, A. J., Creusot, R. J., Su, L., Sheng, D., Hartnett, M., Iwai, H., Soares, L. R., Fathman, C. G. 2008; 129 (2): 195-201

    Abstract

    Whole genome oligo-microarrays were used to characterize age-dependent and tissue-specific changes in gene expression in pancreatic lymph nodes, spleen, and peripheral blood cells, obtained from up to 8 individual NOD mice at 6 different time points (1.5 to 20 weeks of age), compared to NOD.B10 tissue controls. "Milestone Genes" are genes whose expression was significantly changed (approximately 3 fold) as the result of splicing or changes in transcript level. Milestone Genes were identified among genes within type one diabetes (T1D) susceptibility regions (Idd). Milestone Genes showing uniform patterns of changes in expression at various time points were identified, but the patterns of distribution and kinetics of expression were unique to each tissue. Potential T1D candidate genes were identified among Milestone Genes within Idd regions and/or hierarchical clusters. These studies identified tissue- and age-specific changes in gene expression that may play an important role in the inductive or destructive events of T1D.

    View details for DOI 10.1016/j.clim.2008.07.028

    View details for Web of Science ID 000260553600003

    View details for PubMedID 18801706

  • The single subunit transmembrane E3 ligase gene related to anergy in lymphocytes (GRAIL) captures and then ubiquitinates transmembrane proteins across the cell membrane JOURNAL OF BIOLOGICAL CHEMISTRY Lineberry, N., Su, L., Soares, L., Fathman, C. G. 2008; 283 (42): 28497-28505

    Abstract

    The ubiquitin E3 ligase gene related to anergy in lymphocytes (GRAIL) (Rnf128) is a type 1 transmembrane protein that induces T cell anergy through the ubiquitination activity of its cytosolic RING finger. GRAIL also contains an equally large luminal region consisting primarily of an uncharacterized protease-associated (PA) domain. Using two-hybrid technology to screen for proteins that bound the PA domain we identified CD151, a member of the tetraspanin family of membrane proteins. GRAIL bound to the luminal/extracellular portion of both CD151 and the related tetraspanin CD81 using its PA domain, which promoted ubiquitination of cytosolic lysine residues. GRAIL exhibited specificity for lysines only within the tetraspanin amino terminus even in the presence of other cytosolic lysine residues in the substrate. GRAIL-mediated ubiquitination promoted proteasomal degradation and cell surface down-regulation of tetraspanins via Lys-48 linkages. As a result, the juxtaposition of PA and RING finger domains across a lipid bilayer facilitates the capture of transmembrane substrates for subsequent ubiquitination. These findings identify for the first time a single subunit E3 ligase containing a substrate-binding domain spatially restricted by a membrane from its E2 recruitment domain as well as an E3 ligase for members of the tetraspanin family.

    View details for DOI 10.1074/jbc.M805092200

    View details for Web of Science ID 000259969300052

    View details for PubMedID 18713730

  • Cutting edge: The transmembrane E3 ligase GRAIL ubiquitinates the costimulatory molecule CD40 ligand during the induction of T cell anergy JOURNAL OF IMMUNOLOGY Lineberry, N. B., Su, L. L., Lin, J. T., Coffey, G. P., Seroogy, C. M., Fathman, C. G. 2008; 181 (3): 1622-1626

    Abstract

    Activation of naive T lymphocytes is regulated through a series of discrete checkpoints that maintain unresponsiveness to self. During this multistep process, costimulatory interactions act as inducible signals that allow APCs to selectively mobilize T cells against foreign Ags. In this study, we provide evidence that the anergy-associated E3 ubiquitin ligase GRAIL (gene related to anergy in lymphocytes) regulates expression of the costimulatory molecule CD40L on CD4 T cells. Using its luminal protease-associated domain, GRAIL binds to the luminal/extracellular portion of CD40L and facilitates transfer of ubiquitin molecules from the intracellular GRAIL RING (really interesting new gene) finger to the small cytosolic portion of CD40L. Down-regulation of CD40L occurred following ectopic expression of GRAIL in naive T cells from CD40(-/-) mice, and expression of GRAIL in bone marrow chimeric mice was associated with diminished lymphoid follicle formation. These data provide a model for intrinsic T cell regulation of costimulatory molecules and a molecular framework for the initiation of clonal T cell anergy.

    View details for Web of Science ID 000257898100006

    View details for PubMedID 18641297

  • Tissue-targeted therapy of autoimmune diabetes using dendritic cells transduced to express IL-4 in NOD mice CLINICAL IMMUNOLOGY Creusot, R. J., Yaghoubi, S. S., Kodama, K., Dang, D. N., Dang, V. H., Breckpot, K., Thielemans, K., Gambhir, S. S., Fathman, C. G. 2008; 127 (2): 176-187

    Abstract

    A deficit in IL-4 production has been previously reported in both diabetic human patients and non-obese diabetic (NOD) mice. In addition, re-introducing IL-4 into NOD mice systemically, or as a transgene, led to a beneficial outcome in most studies. Here, we show that prediabetic, 12-week old female NOD mice have a deficit in IL-4 expression in the pancreatic lymph nodes (PLN) compared to age-matched diabetes-resistant NOD.B10 mice. By bioluminescence imaging, we demonstrated that the PLN was preferentially targeted by bone marrow-derived dendritic cells (DCs) following intravenous (IV) administration. Following IV injection of DCs transduced to express IL-4 (DC/IL-4) into 12-week old NOD mice, it was possible to significantly delay or prevent the onset of hyperglycemia. We then focused on the PLN to monitor, by microarray analysis, changes in gene expression induced by DC/IL-4 and observed a rapid normalization of the expression of many genes, that were otherwise under-expressed compared to NOD.B10 PLN. The protective effect of DC/IL-4 required both MHC and IL-4 expression by the DCs. Thus, adoptive cellular therapy, using DCs modified to express IL-4, offers an effective, tissue-targeted cellular therapy to prevent diabetes in NOD mice at an advanced stage of pre-diabetes, and may offer a safe approach to consider for treatment of high risk human pre-diabetic patients.

    View details for DOI 10.1016/j.clim.2007.12.009

    View details for Web of Science ID 000255231100010

    View details for PubMedID 18337172

  • Naive and memory T cells induce different types of graft-versus-host disease JOURNAL OF IMMUNOLOGY Dutt, S., Tseng, D., Ermann, J., George, T. I., Liu, Y. P., Davis, C. R., Fathman, C. G., Strober, S. 2007; 179 (10): 6547-6554

    Abstract

    The goal of this study was to compare the ability of donor naive and alloantigen-primed effector memory T cells to induce graft-vs-host disease after bone marrow transplantation in MHC-mismatched irradiated host mice. Purified CD4(+) naive (CD62L(high)CD44(low)) T cells and CD4(+) effector memory (CD62L(low)CD44(high)) T cells obtained from unprimed donors and donors primed to host alloantigens, respectively, were injected into host mice, and the rapidity, severity, and pattern of tissue injury of graft-vs-host disease was assessed. Unexpectedly, the naive T cells induced a more acute and severe colitis than the primed memory cells. Whereas the naive T cells expressing CD62L and CCR7 lymph node homing receptors vigorously expanded in mesenteric lymph nodes and colon by day 6 after transplantation, the primed memory T cells without these receptors had 20- to 100-fold lower accumulation at this early time point. These differences were reflected in the significantly more rapid decline in survival and weight loss induced by naive T cells. The primed memory T cells had a greater capacity to induce chronic colitis and liver injury and secrete IL-2 and IFN-gamma in response to alloantigenic stimulation compared with memory T cells from unprimed donors. Nevertheless, the expected increase in potency as compared with naive T cells was not observed due to differences in the pattern and kinetics of tissue injury.

    View details for Web of Science ID 000250792700021

    View details for PubMedID 17982043

  • Multimodality imaging of T-cell hybridoma trafficking in collagen-induced arthritic mice: image-based estimation of the number of cells accumulating in mouse paws JOURNAL OF BIOMEDICAL OPTICS Yaghoubi, S. S., Creusot, R. J., Ray, P., Fathman, C. G., Gambhir, S. S. 2007; 12 (6)

    Abstract

    Appropriate targeting of therapeutic cells is essential in adoptive cellular gene therapy (ACGT). Imaging cell trafficking in animal models and patients will guide development of ACGT protocols. Collagen type II (C-II)-specific T cell hybridomas are transduced with a lentivirus carrying a triple fusion reporter gene (TFR) construct consisting of a fluorescent reporter gene (RG), a bioluminescent RG (hRluc), and a positron emission tomography (PET) RG. Collagen-induced arthritic (CIA) mice are scanned with a bioluminescence imaging camera before and after implantation of various known cell quantities in their paws. Linear regression analysis yields equations relating two parameters of image signal intensity in mice paws to the quantity of hRluc expressing cells in the paws. Afterward, trafficking of intravenously injected cells is studied by quantitative analysis of bioluminescence images. Comparison of the average cell numbers does not demonstrate consistently higher accumulation of T-cell hybridomas in the paws with higher inflammation scores, and injecting more cells does not cause increased accumulation. MicroPET images illustrate above background signal in the inflamed paws and chest areas of CIA mice. The procedures described in this study can be used to derive equations for cells expressing other bioluminescent RGs and in other animal models.

    View details for DOI 10.1117/1.2821415

    View details for Web of Science ID 000252851100046

    View details for PubMedID 18163841

  • Molecular mechanisms of CD4(+) T-cell anergy NATURE REVIEWS IMMUNOLOGY Fathman, C. G., Lineberry, N. B. 2007; 7 (8): 599-609

    Abstract

    Directing both innate and adaptive immune responses against foreign pathogens with correct timing, location and specificity is a fundamental objective for the immune system. Full activation of CD4+ T cells requires the binding of peptide-MHC complexes coupled with accessory signals provided by the antigen-presenting cell. However, aberrant activation of the T-cell receptor alone in mature T cells can produce a long-lived state of functional unresponsiveness, known as anergy. Recent studies probing both immune signalling pathways and the ubiquitin-proteasome system have helped to refine and elaborate current models for the molecular mechanisms underlying T-cell anergy. Controlling anergy induction and maintenance will be a key component in the future to mitigate unwanted T-cell activation that leads to autoimmune disease.

    View details for DOI 10.1038/nri2131

    View details for Web of Science ID 000248394100012

    View details for PubMedID 17612584

  • A novel E3 ubiquitin ligase substrate screen identifies Rho guanine dissociation inhibitor as a substrate of gene related to anergy in lymphocytes JOURNAL OF IMMUNOLOGY Su, L., Lineberry, N., Huh, Y., Soares, L., Fathman, C. G. 2006; 177 (11): 7559-7566

    Abstract

    Ubiquitination of eukaryotic proteins regulates a broad range of cellular processes, including regulation of T cell activation and tolerance. We have previously demonstrated that gene related to anergy in lymphocytes (GRAIL), a ring finger ubiquitin E3 ligase, is required for the induction of T cell anergy; however, the substrate(s) for GRAIL E3 ligase activity is/are unknown. In this study, we report a novel prokaryotic system developed to screen for substrates of E3 ligases. Using this screen, Rho guanine dissociation inhibitor (RhoGDI) was identified as a potential substrate of GRAIL. GRAIL was subsequently demonstrated to bind and ubiquitinate RhoGDI, although GRAIL-mediated ubiquitination of RhoGDI did not result in proteosomal degradation. Expression of GRAIL in T cells resulted in specific inhibition of RhoA GTPase activation; activation of Rac1, cdc42, and Ras GTPases were not affected. Interestingly, stable T cell lines expressing dominant-negative RhoA mimicked the GRAIL-mediated IL-2 inhibition phenotype, and T cells expressing constitutively active RhoA were able to overcome GRAIL-mediated inhibition of IL-2 expression. These findings validate our prokaryotic screen as a method of identifying substrates for ubiquitin E3 ligases and suggest a role for Rho effector molecules in T cell anergy.

    View details for Web of Science ID 000242261800012

    View details for PubMedID 17114425

  • Does our current understanding of the molecular basis of immune tolerance predict new therapies for autoimmune disease? NATURE CLINICAL PRACTICE RHEUMATOLOGY Tarner, I. H., Fathman, C. G. 2006; 2 (9): 491-499

    Abstract

    The creation of specific immune tolerance has often been referred to as the ultimate goal of immunotherapy, because it would allow autoimmune disease to be reversed without the need for nonspecific and potentially harmful immunosuppressive therapy. Studies performed during the past decade have been immensely fruitful in terms of advances in our understanding of the cellular and molecular mechanisms of immune tolerance, and have paved the way for successful exploitation of these mechanisms for therapeutic purposes. Important developments include an increased understanding of central and peripheral tolerance, and treatment strategies that mimic the mechanisms behind deletion of self-reactive cells, the identification of crucial gene products that are involved in the induction of anergy, and the characterization of regulatory T cells and protocols for their induction and expansion for therapeutic applications. These landmarks of immune-tolerance research are summarized and their potential use in the immunotherapy of autoimmune disease discussed.

    View details for DOI 10.1038/ncprheum0272

    View details for Web of Science ID 000240200400010

    View details for PubMedID 16951704

  • T cell anergy: Where it's LAT IMMUNITY Lineberry, N., Fathman, C. G. 2006; 24 (5): 501-503

    Abstract

    T cell receptor engagement activates selective signaling pathways in T lymphocytes under different conditions. In this issue of Immunity, demonstrate that anergic T cells are selectively defective in LAT activation.

    View details for DOI 10.1016/j.immuni.2006.05.002

    View details for Web of Science ID 000237970100004

    View details for PubMedID 16713965

  • CYLD: deubiquitination-induced TCR signaling NATURE IMMUNOLOGY Lineberry, N., Fathman, C. G. 2006; 7 (4): 369-370

    View details for DOI 10.1038/ni0406-369

    View details for Web of Science ID 000236171000009

    View details for PubMedID 16550202

  • Developing the concept of adoptive cellular gene therapy of rheumatoid arthritis AUTOIMMUNITY REVIEWS Tarner, I. H., Neumann, E., GAY, S., Fathman, C. G., Mulller-Ladner, U. 2006; 5 (2): 148-152

    Abstract

    Progressive destruction of articular cartilage and bone is the pivotal problem of rheumatoid arthritis (RA). Joint destruction is the cause of severe disability and determines the long-term outcome of disease. Conventional therapy does not control this destructive process sufficiently and the anti-rheumatic drugs available today can cause severe systemic adverse effects. Local application of chondroprotective and osteoprotective agents by means of gene therapy would be an attractive alternative to conventional therapy of RA and could provide long-term expression of the therapeutic agents and minimize systemic adverse effects. For this purpose, we have developed the concept of adoptive cellular gene therapy. This treatment strategy is based on using genetically engineered cells that home specifically to sites of autoimmune inflammation and thus allow local delivery of therapeutic gene products. Ex vivo transduction of these cells avoids systemic exposure of the host to the transgene-encoding vector and thus adds to the safety of this approach. In this article of the CIS Spring School in Autoimmune Diseases 2005 proceedings, we review our work on developing the strategy of adoptive cellular gene therapy and summarize recent advances in the evaluation of therapeutic effects and the identification of novel therapeutic targets.

    View details for DOI 10.1016/j.autrev.2005.09.009

    View details for Web of Science ID 000235349400018

    View details for PubMedID 16431349

  • CD4(+)CD25(+) regulatory T cells and their therapeutic potential ANNUAL REVIEW OF MEDICINE Randolph, D. A., Fathman, C. G. 2006; 57: 381-402

    Abstract

    The human immune system mounts specific responses to a vast array of antigens. Although this is clearly beneficial in fighting off harmful infections and cancerous cells, the system must be carefully controlled to ensure that normal self-antigens are not targeted. A recently characterized subset of T cells, identified by their cell surface expression of CD4 and CD25, is critical in regulating the function of other immune cells and preventing potentially harmful autoimmune responses. This article reviews what is currently known about these so-called regulatory T cells and discusses the therapeutic potential of these cells to modulate human immune-based diseases.

    View details for DOI 10.1146/annurev.med.57.121304.131337

    View details for Web of Science ID 000235981900024

    View details for PubMedID 16409156

  • L-selectin and beta(7) integrin on donor CD4 T cells are required for the early migration to host mesenteric lymph nodes and acute colitis of graft-versus-host disease BLOOD Dutt, S., Ermann, J., Tseng, D., Liu, Y. P., George, T. I., Fathman, C. G., Strober, S. 2005; 106 (12): 4009-4015

    Abstract

    The homing receptors L-selectin and alpha4beta7 integrin facilitate entry of T cells into the gut-associated organized lymphoid tissues such as the mesenteric lymph nodes and Peyer patches. We studied the impact of inactivation of genes encoding these receptors on the ability of purified donor CD4+ T cells to induce acute lethal graft-versus-host disease (GVHD) associated with severe colitis in irradiated major histocompatibility complex (MHC)-mismatched mice. Whereas lack of expression of a single receptor had no significant impact on the severity of colitis and GVHD, the lack of expression of both receptors markedly ameliorated colitis and early deaths observed with wild-type (WT) T cells. The changes in colitis and GVHD were reflected in a marked reduction in the early accumulation of donor T cells in the mesenteric lymph nodes and subsequently in the colon. The purified WT donor CD4+ T cells did not accumulate early in the Peyer patches and failed to induce acute injury to the small intestine. In conclusion, the combination of CD62L and beta7 integrin is required to induce acute colitis and facilitate entry of CD4+ donor T cells in the mesenteric nodes associated with lethal GVHD in allogeneic hosts.

    View details for DOI 10.1182/blood-2005-06-2339

    View details for Web of Science ID 000233662400058

    View details for PubMedID 16105972

  • Targeted gene therapy of autoimmune diseases: advances and prospects. Expert review of clinical immunology Creusot, R. J., Fathman, C. G., Müller-Ladner, U., Tarner, I. H. 2005; 1 (3): 385-404

    Abstract

    Idealized gene therapy of autoimmune diseases would mean getting the right drug to the right place at the right time to affect the right mechanism of action. In other words, a specific gene therapy strategy needs to have functional, spatial and temporal specificity. Functional specificity implies targeting the cellular, molecular and/or genetic mechanisms relevant to the disease, without affecting nondiseased organs or tissues through mechanisms that cause adverse effects. Spatial specificity means the delivery of the therapeutic agent exclusively to sites and cells that are relevant to the disease. Temporal specificity is, in principle, synonymous with controlled on-demand expression of the therapeutic gene and thus represents a major safety feature. This article reviews recent advances in strategies to use gene therapy in the treatment of autoimmune diseases.

    View details for DOI 10.1586/1744666X.1.3.385

    View details for PubMedID 20476990

  • Molecular imaging using labeled donor tissues reveals patterns of engraftment, rejection, and survival in transplantation TRANSPLANTATION Cao, Y. A., Bachmann, M. H., Beilhack, A., Yang, Y., Tanaka, M., Swijnenburg, R. J., Reeves, R., Taylor-Edwards, C., Schulz, S., Doyle, T. C., Fathman, C. G., Robbins, R. C., Herzenberg, L. A., Negrin, R. S., Contag, C. H. 2005; 80 (1): 134-139

    Abstract

    Tissue regeneration and transplantation of solid organs involve complex processes that can only be studied in the context of the living organism, and methods of analyzing these processes in vivo are essential for development of effective transplantation and regeneration procedures. We utilized in vivo bioluminescence imaging (BLI) to noninvasively visualize engraftment, survival, and rejection of transplanted tissues from a transgenic donor mouse that constitutively expresses luciferase. Dynamic early events of hematopoietic reconstitution were accessible and engraftment from as few as 200 transplanted whole bone marrow (BM) cells resulted in bioluminescent foci in lethally irradiated, syngeneic recipients. The transplantation of autologous pancreatic Langerhans islets and of allogeneic heart revealed the tempo of transplant degeneration or immune rejection over time. This imaging approach is sensitive and reproducible, permits study of the dynamic range of the entire process of transplantation, and will greatly enhance studies across various disciplines involving transplantation.

    View details for DOI 10.1097/01.TP.0000164347.50559.A3

    View details for Web of Science ID 000230473800023

    View details for PubMedID 16003245

  • An array of possibilities for the study of autoimmunity NATURE Fathman, C. G., Soares, L., Chan, S. M., Utz, P. J. 2005; 435 (7042): 605-611

    Abstract

    Since the completion of the sequencing of the human genome, scientific focus has shifted from studying genes to analysing the much larger number of proteins encoded by them. Several proteins can be generated from a single gene depending on how the genetic information is read (transcribed) and how the resultant protein is modified following translation (post-translational modification). Genomic and proteomic technologies are already providing useful information about autoimmune disease, and they are likely to lead to important discoveries within the next decade.

    View details for DOI 10.1038/nature03726

    View details for Web of Science ID 000229476200038

    View details for PubMedID 15931213

  • Only the CD62L(+) subpopulation of CD4(+)CD25(+) regulatory T cells protects from lethal acute GVHD BLOOD Ermann, J., Hoffmann, P., Edinger, M., Dutt, S., Blankenberg, F. G., Higgins, J. P., Negrin, R. S., Fathman, C. G., Strober, S. 2005; 105 (5): 2220-2226

    Abstract

    CD4+CD25+ regulatory T (Treg) cells are potent modulators of alloimmune responses. In murine models of allogeneic bone marrow transplantation, adoptive transfer of donor CD4+CD25+ Treg cells protects recipient mice from lethal acute graft-versus-host disease (aGVHD) induced by donor CD4+CD25- T cells. Here we examined the differential effect of CD62L+ and CD62L- subsets of CD4+CD25+ Treg cells on aGVHD-related mortality. Both subpopulations showed the characteristic features of CD4+CD25+ Treg cells in vitro and did not induce aGVHD in vivo. However, in cotransfer with donor CD4+CD25- T cells, only the CD62L+ subset of CD4+CD25+ Treg cells prevented severe tissue damage to the colon and protected recipients from lethal aGVHD. Early after transplantation, a higher number of donor-type Treg cells accumulated in host mesenteric lymph node (LN) and spleen when CD4+CD25+CD62L+ Treg cells were transferred compared with the CD62L- subset. Subsequently, CD4+CD25+CD62L+ Treg cells showed a significantly higher capacity than their CD62L- counterpart to inhibit the expansion of donor CD4+CD25- T cells. The ability of Treg cells to efficiently enter the priming sites of pathogenic allo-reactive T cells appears to be a prerequisite for their protective function in aGVHD.

    View details for DOI 10.1182/blood-2004-05-2044

    View details for Web of Science ID 000227308400064

    View details for PubMedID 15546950

  • Protein microarrays for multiplex analysis of signal transduction pathways Nature Medicine Chan, Steven M., Ermann, Joerg, Su, Leon, Fathman, C. Garrison, Utz, Paul J. 2005; 10: 1390-1396
  • Fostering interdisciplinary immunology Clinical Immunology Huston, David P., Utz Paul J., Fathman C. Garrison 2005; 115: 1-2
  • Protein microarrays for multiplex analysis of signal transduction pathways NATURE MEDICINE Chan, S. M., Ermann, J., Su, L., Fathman, C. G., Utz, P. J. 2004; 10 (12): 1390-1396

    Abstract

    We have developed a multiplexed reverse phase protein (RPP) microarray platform for simultaneous monitoring of site-specific phosphorylation of numerous signaling proteins using nanogram amounts of lysates derived from stimulated living cells. We first show the application of RPP microarrays to the study of signaling kinetics and pathway delineation in Jurkat T lymphocytes. RPP microarrays were used to profile the phosphorylation state of 62 signaling components in Jurkat T cells stimulated through their membrane CD3 and CD28 receptors, identifying a previously unrecognized link between CD3 crosslinking and dephosphorylation of Raf-1 at Ser259. Finally, the potential of this technology to analyze rare primary cell populations is shown in a study of differential STAT protein phosphorylation in interleukin (IL)-2-stimulated CD4(+)CD25(+) regulatory T cells. RPP microarrays, prepared using simple procedures and standard microarray equipment, represent a powerful new tool for the study of signal transduction in both health and disease.

    View details for DOI 10.1038/nm1139

    View details for Web of Science ID 000225500900035

    View details for PubMedID 15558056

  • Murine CD4(+) CD25(+) regulatory T cells fail to undergo chromatin remodeling across the proximal promoter region of the IL-2 gene JOURNAL OF IMMUNOLOGY Su, L., Creusot, R. J., Gallo, E. M., Chan, S. M., Utz, P. J., Fathman, C. G., Ermann, J. 2004; 173 (8): 4994-5001

    Abstract

    CD4+CD25+ regulatory T cells (Treg) acquire unique immunosuppressive properties while maintaining an anergy phenotype when activated in vitro under conditions that induce IL-2 production and proliferation in conventional CD4+ T cells. We investigated the mechanism underlying one component of this naturally anergic phenotype, the inability of the Treg cells to produce IL-2 following activation. Analysis of freshly isolated murine CD4+CD25+ Treg and conventional CD4+CD25- T cells following PMA/ionomycin stimulation demonstrated no differences in inducible AP-1 formation, an important transcriptional complex in regulating IL-2 gene expression. Although p38 MAPK and ERK1/2 protein kinases were phosphorylated with similar kinetics, we observed diminished activation of JNK in the CD4+CD25+ Treg cells. However, lentiviral-mediated reconstitution of the JNK pathway using a constitutively active construct did not overcome the block in IL-2 synthesis. Using a PCR-based chromatin accessibility assay we found that the minimal IL-2 promoter region of CD4+CD25+ Treg cells, unlike conventional CD4 T cells, did not undergo chromatin remodeling following stimulation, suggesting that the inability of CD4+CD25+ Treg cells to secrete IL-2 following activation is controlled at the chromatin level.

    View details for Web of Science ID 000224392200028

    View details for PubMedID 15470042

  • Gene therapy for type 1 diabetes: a novel approach for targeted treatment of autoimmunity JOURNAL OF CLINICAL INVESTIGATION Creusot, R. J., Fathman, C. G. 2004; 114 (7): 892-894

    Abstract

    It has been difficult to develop therapies that target those T cells initiating and mediating the pathogenesis of autoimmune disease. Indeed, most current treatments indiscriminately affect both the autoreactive T cells and the "good" T cells, putting the patient at risk of compromised immune function. A new approach raises the possibility of targeted therapy for autoimmunity. Transplantation of hematopoietic stem cells modified to express a protective form of MHC class II corrects a defect in central tolerance. This method contrasts with other targeted therapies that attempt to modify peripheral tolerance, which is also defective in type 1 diabetes mellitus.

    View details for DOI 10.1172/JC1200423168

    View details for Web of Science ID 000224223800008

    View details for PubMedID 15467826

  • The gene related to anergy in lymphocytes, an E3 ubiquitin ligase, is necessary for anergy induction in CD4 T cells JOURNAL OF IMMUNOLOGY Seroogy, C. M., Soares, L., Ranheim, E. A., Su, L., Holness, C., Bloom, D., Fathman, C. G. 2004; 173 (1): 79-85

    Abstract

    Acquisition of the anergy phenotype in T cells is blocked by inhibitors of protein synthesis and calcineurin activity, suggesting that anergic T cells may have a unique genetic program. Retroviral transduction of hemopoietic stem cells from TCR transgenic mice and subsequent reconstitution of syngeneic mice to express the E3 ubiquitin ligase, gene related to anergy in lymphocytes (GRAIL), or an enzymatically inactive form, H2N2 GRAIL, allowed analysis of the role of GRAIL in T cell anergy in vivo. Constitutive expression of GRAIL was sufficient to render naive CD4 T cells anergic, however, when the enzymatically inactive form H2N2 GRAIL was expressed, it functioned as a dominant negative of endogenous GRAIL and blocked the development of anergy. These data provide direct evidence that a biochemical pathway composed of GRAIL and/or GRAIL-interacting proteins is important in the development of the CD4 T cell anergic phenotype in vivo.

    View details for Web of Science ID 000222170900014

    View details for PubMedID 15210761

  • Targeted gene therapy: frontiers in the development of 'smart drugs' TRENDS IN BIOTECHNOLOGY Tarner, I. H., Muller-Ladner, U., Fathman, C. G. 2004; 22 (6): 304-310

    Abstract

    Chronic diseases, particularly malignancies and immune-mediated inflammatory diseases (IMIDs), are a challenging frontier for clinical diagnosis and treatment, as well as for biomedical research. Current treatment regimens are frequently insufficient and thus new treatment strategies are needed. Novel therapies for disabling such diseases should provide improvements with respect to safety, efficacy and cost. To fulfill these three key criteria, recent research efforts have focused on the development of 'smart drugs'. This review highlights some examples of the rapidly expanding possibilities that current biotechnology has to offer in the development of novel therapeutic strategies for complex diseases such as IMIDs. Special attention is given to advances in, and limitations of, controlled and targeted gene product application in inflammatory diseases.

    View details for Web of Science ID 000222301000011

    View details for PubMedID 15158060

  • Two isoforms of otubain 1 regulate T cell anergy via GRAIL NATURE IMMUNOLOGY Soares, L., Seroogy, C., Skrenta, H., Anandasabapathy, N., Lovelace, P., Chung, C. D., Engleman, E., Fathman, C. G. 2004; 5 (1): 45-54

    Abstract

    The active ubiquitin E3 ligase GRAIL is crucial in the induction of CD4 T cell anergy. Here we show that GRAIL is associated with and regulated by two isoforms of the ubiquitin-specific protease otubain 1. In lethally irradiated mice reconstituted with bone marrow cells from T cell receptor-transgenic mice retrovirally transduced to express the genes encoding these proteases, otubain 1-expressing cells contained negligible amounts of endogenous GRAIL, proliferated well and produced large amounts of interleukin 2 after antigenic stimulation. In contrast, cells expressing the alternatively spliced isoform, otubain 1 alternative reading frame 1, contained large amounts of endogenous GRAIL and were functionally anergic, and they proliferated poorly and produced undetectable interleukin 2 when stimulated in a similar way. Thus, these two proteins have opposing epistatic functions in controlling the stability of GRAIL expression and the resultant anergy phenotype in T cells.

    View details for DOI 10.1038/ni1017

    View details for Web of Science ID 000187635800013

    View details for PubMedID 14661020

  • Costimulatory signals controlling regulatory T cells PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Ermann, J., Fathman, C. G. 2003; 100 (26): 15292-15293

    View details for DOI 10.1073/pnas.0307001100

    View details for Web of Science ID 000187554600003

    View details for PubMedID 14676329

  • CD4(+)CD25(+) regulatory T cells preserve graft-versus-tumor activity while inhibiting graft-versus-host disease after bone marrow transplantation NATURE MEDICINE Edinger, M., Hoffmann, P., Ermann, J., Drago, K., Fathman, C. G., Strober, S., Negrin, R. S. 2003; 9 (9): 1144-1150

    Abstract

    Mature donor T cells cause graft-versus-host disease (GVHD), but they are also the main mediators of the beneficial graft-versus-tumor (GVT) activity of allogeneic bone marrow transplantation. Suppression of GVHD with maintenance of GVT activity is a desirable outcome for clinical transplantation. We have previously shown that donor-derived CD4+CD25+ regulatory T cells inhibit lethal GVHD after allogeneic bone marrow transplantation across major histocompatibility complex (MHC) class I and II barriers in mice. Here we demonstrate that in host mice with leukemia and lymphoma, CD4+CD25+ regulatory T cells suppress the early expansion of alloreactive donor T cells, their interleukin-2-receptor (IL-2R) alpha-chain expression and their capacity to induce GVHD without abrogating their GVT effector function, mediated primarily by the perforin lysis pathway. Thus, CD4+CD25+ T cells are potent regulatory cells that can separate GVHD from GVT activity mediated by conventional donor T cells.

    View details for DOI 10.1038/nm915

    View details for Web of Science ID 000185061600026

    View details for PubMedID 12925844

  • Localized expression of an anti-TNF single-chain antibody prevents development of collagen-induced arthritis GENE THERAPY Smith, R., Tarner, I. H., Hollenhorst, M., Lin, C., Levicnik, A. U., Fathman, C. G., Nolan, G. P. 2003; 10 (15): 1248-1257

    Abstract

    Although systemic administration of neutralizing anti-TNF antibodies has been used successfully in treating rheumatoid arthritis, there is a potential for side effects. We transduced a collagen reactive T-cell hybridoma with tissue-specific homing properties to assess therapeutic effects of local delivery to inflamed joints of anti-TNF single-chain antibodies (scFv) by adoptive cellular gene therapy. Cell culture medium conditioned with 1 x 10(6) scFv producer cells/ml had TNF neutralizing capacity in vitro equivalent to 50 ng/ml anti-TNF monoclonal antibody. Adding a kappa chain constant domain to the basic scFv (construct TN3-Ckappa) gave increased in vitro stability and in vivo therapeutic effect. TN3-Ckappa blocked development of collagen-induced arthritis in DBA/1LacJ mice for >60 days. Transgene expression was detected in the paws but not the spleen of treated animals for up to 55 days postinjection. No significant variations in cell proliferation or cytokine secretion were found in splenocytes or peripheral lymphocytes. IL-6 expression was blocked in the diseased paws of mice in the scFv treatment groups compared to controls. In conclusion, we have shown that local expression of an anti-inflammatory agent blocks disease development without causing demonstrable systemic immune function changes. This is encouraging for the potential development of safe adoptive cellular therapies to treat autoimmunity.

    View details for DOI 10.1038/sj.gt.3301980

    View details for Web of Science ID 000184207000007

    View details for PubMedID 12858190

  • GRAIL: An E3 ubiquitin ligase that inhibits cytokine gene transcription is expressed in anergic CD4(+) T cells IMMUNITY Anandasabapathy, N., Ford, G. S., Bloom, D., Holness, C., Paragas, V., Seroogy, C., Skrenta, H., Hollenhorst, M., Fathman, C. G., Soares, L. 2003; 18 (4): 535-547

    Abstract

    T cell anergy may serve to limit autoreactive T cell responses. We examined early changes in gene expression after antigen-TCR signaling in the presence (activation) or absence (anergy) of B7 costimulation. Induced expression of GRAIL (gene related to anergy in lymphocytes) was observed in anergic CD4(+) T cells. GRAIL is a type I transmembrane protein that localizes to the endocytic pathway and bears homology to RING zinc-finger proteins. Ubiquitination studies in vitro support GRAIL function as an E3 ubiquitin ligase. Expression of GRAIL in retrovirally transduced T cell hybridomas dramatically limits activation-induced IL-2 and IL-4 production. Additional studies suggest that GRAIL E3 ubiquitin ligase activity and intact endocytic trafficking are critical for cytokine transcriptional regulation. Expression of GRAIL after an anergizing stimulus may result in ubiquitin-mediated regulation of proteins essential for mitogenic cytokine expression, thus positioning GRAIL as a key player in the induction of the anergic phenotype.

    View details for Web of Science ID 000182353900009

    View details for PubMedID 12705856

  • Treatment of autoimmune disease by adoptive cellular gene therapy MYASTHENIA GRAVIS AND RELATED DISORDERS Tarner, I. H., Slavin, A. J., McBride, J., Levicnik, A., Smith, R., Nolan, G. P., Contag, C. H., Fathman, C. G. 2003; 998: 512-519

    Abstract

    Autoimmune disorders represent inappropriate immune responses directed at self-tissue. Antigen-specific CD4+ T cells and antigen-presenting dendritic cells (DCs) are important mediators in the pathogenesis of auto-immune disease and thus are ideal candidates for adoptive cellular gene therapy, an ex vivo approach to therapeutic gene transfer. Using retrovirally transduced cells and luciferase bioluminescence, we have demonstrated that primary T cells, T cell hybridomas, and DCs rapidly and preferentially home to the sites of inflammation in animal models of multiple sclerosis, arthritis, and diabetes. These cells, transduced with retroviral vectors to drive expression of various "regulatory proteins" such as IL-4, IL-10, IL-12p40, and anti-TNF scFv, deliver these immunoregulatory proteins to the inflamed lesions, providing therapy for experimental autoimmune encephalitis (EAE), collagen-induced arthritis (CIA), and nonobese diabetic mice (NOD).

    View details for Web of Science ID 000186107400066

    View details for PubMedID 14592922

  • Scratching the (T cell) surface. Genome biology Ermann, J., Chung, C. D., Fathman, C. G. 2003; 5 (1): 202-?

    Abstract

    Using a genome-scale approach to study transcription levels in a human CD8+ T-cell clone, a recent study has suggested that the repertoire of molecules on the surface of T cells is close to being completely characterized.

    View details for PubMedID 14709166

  • T-cell anergy - From phenotype to genotype and back IMMUNOLOGIC RESEARCH Seroogy, C. M., Fathman, C. G. 2003; 28 (3): 255-264

    Abstract

    For many decades, anergy has been used as a descriptive term to describe a state of antigen-specific unresponsiveness. A better understanding of this phenotype was revealed in the 1980s using in vitro model systems. These model systems demonstrated that protein synthesis and mobilization of Ca2+ was required leading to the pursuit of a novel gene(s) that would be unique to the anergy phenotype. Several putative "anergy factors" have been suggested. In this review, we provide an overview of the anergy phenotype and proposed anergy-related genes. To date, no single gene has been described that would completely fulfill the criteria of an "anergy factor." We review work from our laboratory describing a novel gene that we have termed Gene Related to Anergy In Lymphocytes (GRAIL) that is upregulated in T cells anergized in vitro and in vivo and, following transduction into T cells, reiterates the anergy phenotype.

    View details for Web of Science ID 000187357900007

    View details for PubMedID 14713718

  • Short polymers of arginine rapidly translocate into vascular cells - Effects on nitric oxide synthesis CIRCULATION JOURNAL Uemura, S., Rothbard, J. B., Matsushita, H., Tsao, P. S., Fathman, C. G., Cooke, J. P. 2002; 66 (12): 1155-1160

    Abstract

    The present study was designed to determine the efficiency of translocation of short polymers of arginine into vascular smooth muscle cells (VSMC) and to determine their effect on nitric oxide (NO) synthesis. Immunostaining revealed that heptamers of L-arginine (R7) rapidly translocated into the VSMC. This rapid transport was not observed with shorter polymers of L-arginine (R5) nor heptamers of lysine (K7). Translocation of R7 was not inhibited by the addition of free L-arginine into the media. When cells were transiently pretreated with R7 or a nonamer of arginine (R9), NO(2) production from cytokine stimulated VSMC was significantly increased, whereas incubation with R5 and K7 had no effect. Short polymers of arginine not only have a unique ability of rapid VSMC translocation but once internalized enhance NO production. Heptamers (or larger polypeptides) of arginine may be useful in therapy to enhance NO production in the vascular system.

    View details for Web of Science ID 000179496000015

    View details for PubMedID 12499624

  • Retroviral gene therapy of collagen-induced arthritis by local delivery of IL-4 CLINICAL IMMUNOLOGY Tarner, I. H., Nakajima, A., Seroogy, C. M., Ermann, J., Levicnik, A., Contag, C. H., Fathman, C. G. 2002; 105 (3): 304-314

    Abstract

    Rheumatoid arthritis (RA) is an autoimmune arthritis, for which treatment options remain limited. This study investigated the potential role of adoptive cellular gene therapy as a novel means for treating the RA animal model collagen-induced arthritis (CIA). Adoptive transfer of antigen-specific T-cell hybridomas retrovirally transduced to express IL-4 1 day before booster immunization significantly reduced the number of inflamed joints. Cell transfer after clinical onset of disease had no therapeutic effect. Bioluminescence imaging showed that the hybridomas migrated to the inflamed joints, thus delivering the regulatory protein locally at the site of inflammation. The homing was, at least in part, due to chemotaxis in response to proinflammatory chemokines that are expressed in inflamed joints. There were no significant changes in the cytokine milieu of the draining lymph nodes, nor in the systemic levels of anti-collagen antibodies in treated mice. We conclude that the beneficial clinical effects observed in our model were most likely based on the local action(s) of IL-4 in the inflamed joints and that the local delivery (and effects) of regulatory cytokines, like IL-4, constitutes a novel and effective method of preventing organ-specific autoimmune disease and of minimizing systemic adverse effects.

    View details for DOI 10.1006/clim.2002.5299

    View details for Web of Science ID 000180187600008

    View details for PubMedID 12498812

  • CD4 T-helper cells engineered to produce IL-10 prevent allergen-induced airway hyperreactivity and inflammation JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY Oh, J. W., Seroogy, C. M., Meyer, E. H., Akbari, O., Berry, G., Fathman, C. G., DeKruyff, R. H., Umetsu, D. T. 2002; 110 (3): 460-468

    Abstract

    T(H)2 cells play a critical role in the pathogenesis of asthma, but the precise immunologic mechanisms that inhibit T(H)2 cell function in vivo are not well understood.The purpose of our studies was to determine whether T cells producing IL-10 regulate the development of asthma.We used gene therapy to generate ovalbumin-specific CD4 T-helper cells to express IL-10, and we examined their capacity to regulate allergen-induced airway hyperreactivity.We demonstrated that the CD4 T-helper cells engineered to express IL-10 abolished airway hyperreactivity and airway eosinophilia in BALB/c mice sensitized and challenged with ovalbumin and in SCID mice reconstituted with ovalbumin-specific T(H)2 effector cells. The inhibitory effect of the IL-10-secreting T-helper cells was accompanied by the presence of increased quantities of IL-10 in the bronchoalveolar lavage fluid, was antigen-specific, and was reversed by neutralization of IL-10. Moreover, neutralization of IL-10 by administration of anti-IL-10 mAb in mice sensitized and challenged with ovalbumin seriously exacerbated airway hyperreactivity and airway inflammation.Our results demonstrate that T cells secreting IL-10 in the respiratory mucosa can indeed regulate T(H)2-induced airway hyperreactivity and inflammation, and they strongly suggest that IL-10 plays an important inhibitory role in allergic asthma.

    View details for DOI 10.1067/mai.2002.127512

    View details for Web of Science ID 000177936900018

    View details for PubMedID 12209095

  • The potential for gene therapy in the treatment of autoimmune disease CLINICAL IMMUNOLOGY Tarner, I. H., Fathman, C. G. 2002; 104 (3): 204-216

    View details for DOI 10.1006/clim.2002.5235

    View details for Web of Science ID 000178549100002

    View details for PubMedID 12217329

  • The Subpopulation of CD4(+) CD25(+) splenocytes that delays adoptive transfer of diabetes expresses L-selectin and high levels of CCR7 JOURNAL OF IMMUNOLOGY Szanya, V., Ermann, J., TAYLOR, C., Holness, C., Fathman, C. G. 2002; 169 (5): 2461-2465

    Abstract

    Recently, CD4(+)CD25(+) T cells have been implicated in the control of diabetes, suggesting that the inflamed islets of Langerhans in prediabetic NOD mice are under peripheral immune surveillance. Here we show that CD4(+)CD25(+) splenocytes inhibit diabetes in cotransfer with islet-infiltrating cells. Furthermore, CD62L expression is necessary for this disease-delaying effect of CD4(+)CD25(+) cells in vivo, but not for their suppressor function in vitro. We demonstrate that the CD4(+)CD25(+)CD62L(+) splenocytes express CCR7 at high levels and migrate toward secondary lymphoid tissue chemokine and ELC (macrophage-inflammatory protein-3beta), lymphoid chemokines, whereas CD4(+)CD25(+)CD62L(-) splenocytes preferentially express CCR2, CCR4, and CXCR3 and migrate toward the corresponding inflammatory chemokines. These data demonstrate that CD4(+)CD25(+)CD62L(+), but not CD4(+)CD25(+)CD62L(-), splenocytes delay diabetes transfer, and that CD4(+)CD25(+) suppressor T cells are comprised of at least two subpopulations that behave differently in cotransfer in vivo and express distinct chemokine receptor and chemotactic response profiles despite demonstrating equivalent suppressor functions in vitro.

    View details for Web of Science ID 000177594100030

    View details for PubMedID 12193715

  • Donor-type CD4(+)CD25(+) regulatory T cells suppress lethal acute graft-versus-host disease after allogeneic bone marrow transplantation JOURNAL OF EXPERIMENTAL MEDICINE Hoffmann, P., Ermann, J., Edinger, M., Fathman, C. G., Strober, S. 2002; 196 (3): 389-399

    Abstract

    Acute graft-versus-host disease (aGVHD) is still a major obstacle in clinical allogeneic bone marrow (BM) transplantation. CD4(+)CD25(+) regulatory T (T(reg)) cells have recently been shown to suppress proliferative responses of CD4(+)CD25(-) T cells to alloantigenic stimulation in vitro and are required for ex vivo tolerization of donor T cells, which results in their reduced potential to induce aGVHD. Here we show that CD4(+)CD25(+) T cells isolated from the spleen or BM of donor C57BL/6 (H-2(b)) mice that have not been tolerized are still potent inhibitors of the alloresponse in vitro and of lethal aGVHD induced by C57BL/6 CD4(+)CD25(-) T cells in irradiated BALB/c (H-2(d)) hosts in vivo. The addition of the CD4(+)CD25(+) T(reg) cells at a 1:1 ratio with responder/inducer CD4(+)CD25(-) T cells resulted in a >90% inhibition of the mixed leukocyte reaction and marked protection from lethal GVHD. This protective effect depended in part on the ability of the transferred CD4(+)CD25(+) T cells to secrete interleukin 10 and occurred if the T(reg) cells were of donor, but not host, origin. Our results demonstrate that the balance of donor-type CD4(+)CD25(+) T(reg) and conventional CD4(+)CD25(-) T cells can determine the outcome of aGVHD.

    View details for DOI 10.1084/jem.20020399

    View details for Web of Science ID 000177658200011

    View details for PubMedID 12163567

  • Adoptive cellular gene therapy of autoimmune disease. Autoimmunity reviews Slavin, A. J., Tarner, I. H., Nakajima, A., Urbanek-Ruiz, I., McBride, J., Contag, C. H., Fathman, C. G. 2002; 1 (4): 213-219

    Abstract

    Autoimmune disorders represent inappropriate immune responses directed at self-tissue. Because CD4+ T cells are important mediators in the pathogenesis of autoimmune disease, they are ideal candidates for cell-based gene therapy. Using retrovirally-transduced cells and luciferase bioluminescence, we have demonstrated that primary T cells and hybridomas, rapidly and preferentially home to the sites of inflammation in organ-specific autoimmune disease. These cells, transduced with retroviral vectors to drive expression of various 'regulatory proteins', such as IL-4, IL-10 and IL-12p40, deliver these immunoregulatory proteins to the inflamed lesions, providing therapy for experimental models of autoimmune disease such as EAE, CIA and NOD mice. This technique was originally developed in our lab in the murine model of multiple sclerosis, EAE, where T cell hybridomas reactive with myelin basic protein (MBP) were transduced to express and used to deliver the modulatory cytokine, IL-4. Recently we have observed that the cytokine receptor antagonist, IL-12p40 transduced anti-myelin basic protein (MBP) TCR-transgenic T cells (but not CII-reactive T cells) were effective in preventing EAE whereas the CII-reactive, but not MBP-reactive T cells, transduced to express IL-12p40, would treat CIA.

    View details for PubMedID 12848998

  • Immunomodulatory vaccination in autoimmune disease ENDOCRINOLOGY AND METABOLISM CLINICS OF NORTH AMERICA Urbanek-Ruiz, I., Ruiz, P. J., Steinman, L., Fathman, C. G. 2002; 31 (2): 441-?

    Abstract

    The development of vaccines is arguably the most significant achievement in medicine to date. The practice of innoculation with the fluid from a sore to protect from a disease actually dates back to ancient China; however, with the introduction of Jenner's smallpox vaccine, and greater understanding of the immune system, vaccines have become specific and systematic. Traditional vaccines have used killed pathogens (hepatitis A and the Salk polio vaccines), immunogenic subunits of a given pathogen (hepatitis B subunit vaccine), or live attenuated pathogens (measles, mumps, rubella, Sabin polio vaccines) to generate protective immunity. Currently, a new generation of vaccines that use the genetic material of a pathogen to elicit protective immunity are being developed. Although the most widespread and successful use of vaccines today remains in the arena of infectious diseases, manipulations of immune responses to protect against cancers, neurologic diseases, and autoimmunity are being explored rigorously.

    View details for Web of Science ID 000176324600012

    View details for PubMedID 12092460

  • A complicated relationship: fulfilling the interactive needs of the T lymphocyte and the dendritic cell. pharmacogenomics journal Mcbride, J. M., Fathman, C. G. 2002; 2 (6): 367-376

    Abstract

    T cells recognize antigenic peptides displayed on the surface of MHC-bearing antigen-presenting cells (APCs), and with sufficient costimulation become activated. However, the ability of an APC (even bearing the correct peptide) to initiate and fulfill the requirements for T cell activation is not easily achieved. Naive T cells use multiple copies of a single receptor to survey the vast array of peptides presented on an APC, and require multiple receptor engagements to initiate T cell activation. Dendritic cells (DCs) are specialized cells with optimal capabilities for priming naive CD4+ T cells. Activation occurs, after initial antigen recognition by T cells, followed by a rapid dialogue between the T cells and the DCs. The resulting changes in both the cytoskeleton and the expression or regulation of cell-surface molecules on both cell types act to further strengthen engagement. In this report, we review the fundamentals of CD4+ T helper cell : DC interactions and discuss recent data concerning the molecular characteristics of this engagement.

    View details for PubMedID 12629502

  • Bioluminescence imaging of lymphocyte trafficking in vivo EXPERIMENTAL HEMATOLOGY Hardy, J., Edinger, M., Bachmann, M. H., Negrin, R. S., Fathman, C. G., Contag, C. H. 2001; 29 (12): 1353-1360

    Abstract

    Lymphocytes are highly mobile cells that travel throughout the body in response to a tremendous variety of stimuli. Revealing lymphocyte trafficking patterns in vivo is necessary for a complete understanding of immune function, as well as cell-cell and cell-tissue interactions in immune development and in response to insult. Although the location of cell populations in various tissues at any given point in time may be revealed by techniques such as flow cytometry and immunofluorescence, these methods are not readily amenable to the assessment of dynamic cell migration patterns in vivo. In the past 5 years, technologies for imaging molecular and cellular changes in living animals have advanced to a point where it is possible to reveal the migratory paths of these vitally important cells. Here, we review one advancement in cellular imaging, in vivo bioluminescence imaging, which addresses the problem of lymphocyte tracking. This imaging strategy has the potential to elucidate the temporal patterns of immune responses and the spatial distribution of lymphocytes within the body.

    View details for Web of Science ID 000172949100001

    View details for PubMedID 11750093

  • Gene therapy in autoimmune disease CURRENT OPINION IN IMMUNOLOGY Tarner, I. H., Fathman, C. G. 2001; 13 (6): 676-682

    Abstract

    Recent work on gene therapies for autoimmune disease has continued to provide insight into the pathogenesis of autoimmunity. Reliable, effective and targeted gene therapy applications have been achieved by using transduced dendritic cells and antigen-specific T cells as delivery vehicles. Bioluminescence imaging has been implemented to visualize cell trafficking and homing in vivo. As a first step into human gene therapy, a phase I clinical trial for assessing the feasibility and safety of gene transfer has been completed in a group of rheumatoid arthritis patients.

    View details for Web of Science ID 000171797600008

    View details for PubMedID 11677089

  • CD4(+)CD25(+) T cells facilitate the induction of T cell anergy JOURNAL OF IMMUNOLOGY Ermann, J., Szanya, V., Ford, G. S., Paragas, V., Fathman, C. G., Lejon, K. 2001; 167 (8): 4271-4275

    Abstract

    T cell anergy is characterized by the inability of the T cell to produce IL-2 and proliferate. It is reversible by the addition of exogenous IL-2. A similar state of unresponsiveness is observed when the proliferative response of murine CD4(+)CD25(-) T cells is suppressed in vitro by coactivated CD4(+)CD25(+) T cells. We have developed a suppression system that uses beads coated with anti-CD3 and anti-CD28 Abs as surrogate APCs to study the interaction of CD4(+)CD25(+) and CD4(+)CD25(-) T cells in vitro. CD4(+)CD25(+) T cell-induced suppression, in this model, was not abrogated by blocking the B7-CTLA-4 pathway. When the CD4(+)CD25(-) T cells were separated from the CD4(+)CD25(+) suppressor cells after 24 h of coactivation by the Ab-coated beads, the CD4(+)CD25(-) T cells were unable to proliferate or to produce IL-2 upon restimulation. The induction of this anergic phenotype in the CD4(+)CD25(-) T cells correlated with the up-regulated expression of the gene related to anergy in lymphocytes (GRAIL), a novel anergy-related gene that acts as a negative regulator of IL-2 transcription. This system constitutes a novel mechanism of anergy induction in the presence of costimulation.

    View details for Web of Science ID 000171858300018

    View details for PubMedID 11591749

  • Autoimmune diseases: genes, bugs and failed regulation NATURE IMMUNOLOGY Ermann, J., Fathman, C. G. 2001; 2 (9): 759-761

    View details for Web of Science ID 000170781200003

    View details for PubMedID 11526377

  • Adoptive immunotherapy of experimental autoimmune encephalomyelitis via T cell delivery of the IL-12 p40 subunit JOURNAL OF IMMUNOLOGY Costa, G. L., Sandora, M. R., Nakajima, A., Nguyen, E. V., Taylor-Edwards, C., Slavin, A. J., Contag, C. H., Fathman, C. G., Benson, J. M. 2001; 167 (4): 2379-2387

    Abstract

    CD4+ T cells are believed to play a central role in the initiation and perpetuation of autoimmune diseases such as multiple sclerosis. In the murine model for multiple sclerosis, experimental autoimmune encephalomyelitis, pathogenic T cells exhibit a Th1-like phenotype characterized by heightened expression of proinflammatory cytokines. Systemic administration of "regulatory" cytokines, which serve to counter Th1 effects, has been shown to ameliorate autoimmune responses. However, the inherent problems of nonspecific toxicity limit the usefulness of systemic cytokine delivery as a potential therapy. Therefore, we used the site-specific trafficking properties of autoantigen-reactive CD4+ T cells to develop an adoptive immunotherapy protocol that provided local delivery of a Th1 cytokine antagonist, the p40 subunit of IL-12. In vitro analysis demonstrated that IL-12 p40 suppressed IFN-gamma production in developing and effector Th1 populations, indicating its potential to modulate Th1-promoted inflammation. We have previously demonstrated that transduction of myelin basic protein-specific CD4+ T cells with pGC retroviral vectors can result in efficient and stable transgene expression. Therefore, we adoptively transferred myelin basic protein-specific CD4+ T cells transduced to express IL-12 p40 into mice immunized to develop experimental autoimmune encephalomyelitis and demonstrated a significant reduction in clinical disease. In vivo tracking of bioluminescent lymphocytes, transduced to express luciferase, using low-light imaging cameras demonstrated that transduced CD4+ T cells trafficked to the central nervous system, where histological analysis confirmed long-term transgene expression. These studies have demonstrated that retrovirally transduced autoantigen-specific CD4+ T cells inhibited inflammation and promoted immunotherapy of autoimmune disorders.

    View details for Web of Science ID 000170949600070

    View details for PubMedID 11490028

  • Immunization with DNA encoding an immunodominant peptide of insulin prevents diabetes in NOD mice CLINICAL IMMUNOLOGY Urbanek-Ruiz, I., Ruiz, P. J., Paragas, V., Garren, H., Steinman, L., Fathman, C. G. 2001; 100 (2): 164-171

    Abstract

    DNA vaccination is an effective means of protecting experimental animals against infectious pathogens and cancer and has more recently been used to prevent autoimmune disease. Insulin-dependent diabetes mellitus (IDDM) is an autoimmune disease characterized by T-cell-mediated destruction of the insulin-secreting beta cells in the pancreas. The NOD mouse is an animal model of IDDM in which several autoantigens, including insulin, have been identified. In this study we demonstrate that vaccination of NOD mice with DNA encoding an immunodominant peptide of insulin (residues 9-23 of the B chain) protects the animals from developing diabetes. Animals injected intramuscularly with a bacterial plasmid encoding the insulin B chain peptide show significantly lower disease incidence and delayed onset of disease when compared to controls. Protection appears to be mediated by insulin B (9-23)-specific down-regulation of IFN-gamma. Our results confirm that DNA vaccination has a protective effect on autoimmunity, the understanding of which will reveal new insights into the immune system and open doors for novel therapies.

    View details for DOI 10.1006/clim.2001.5055

    View details for Web of Science ID 000170304800006

    View details for PubMedID 11465945

  • Antigen-specific T cell-mediated gene therapy in collagen-induced arthritis JOURNAL OF CLINICAL INVESTIGATION Nakajima, A., Seroogy, C. M., Sandora, M. R., Tarner, I. H., Costa, G. L., Taylor-Edwards, C., Bachmann, M. H., Contag, C. H., Fathman, C. G. 2001; 107 (10): 1293-1301

    Abstract

    Autoantigen-specific T cells have tissue-specific homing properties, suggesting that these cells may be ideal vehicles for the local delivery of immunoregulatory molecules. We tested this hypothesis by using type II collagen-specific (CII-specific) CD4(+) T hybridomas or primary CD4(+) T cells after gene transfer, as vehicles to deliver an immunoregulatory protein for the treatment of collagen-induced arthritis (CIA), a mouse model of rheumatoid arthritis (RA). CII-specific T cells or hybridomas were transduced using retroviral vectors to constitutively express the IL-12 antagonist, IL-12 p40. Transfer of engineered CD4(+) T cells after immunization significantly inhibited the development of CIA, while cells transduced with vector control had no effect. The beneficial effect on CIA of IL-12 p40-transduced T cells required TCR specificity against CII, since transfer of T cells specific for another antigen producing equivalent amounts of IL-12 p40 had no effect. In vivo cell detection using bioluminescent labels and RT-PCR showed that transferred CII-reactive T-cell hybridomas accumulated in inflamed joints in mice with CIA. These results indicate that the local delivery of IL-12 p40 by T cells inhibited CIA by suppressing autoimmune responses at the site of inflammation. Modifying antigen-specific T cells by retroviral transduction for local expression of immunoregulatory proteins thus offers a promising strategy for treating RA.

    View details for Web of Science ID 000168867400014

    View details for PubMedID 11375419

  • T cell receptor (TCR)-mediated repertoire selection and loss of TCR V beta diversity during the initiation of a CD4(+) T cell response in vivo JOURNAL OF EXPERIMENTAL MEDICINE Fasso, M., Anandasabapathy, N., Crawford, F., Kappler, J., Fathman, C. G., Ridgway, W. M. 2000; 192 (12): 1719-1730

    Abstract

    We recently described a novel way to isolate populations of antigen-reactive CD4(+) T cells with a wide range of reactivity to a specific antigen, using immunization with a fixed dose of nominal antigen and FACS((R)) sorting by CD4(high) expression. Phenotypic, FACS((R)), functional, antibody inhibition, and major histocompatibility complex-peptide tetramer analyses, as well as T cell receptor Vbeta sequence analyses, of the antigen-specific CD4(high) T cell populations demonstrated that a diverse sperm whale myoglobin 110-121-reactive CD4(+) T cell repertoire was activated at the beginning (day 3 after immunization) of the immune response. Within 6 d of immunization, lower affinity clones were lost from the responding population, leaving an expanded population of oligoclonal, intermediate affinity (and residual high affinity) T cells. This T cell subset persisted for at least 4 wk after immunization and dominated the secondary immune response. These data provide evidence that CD4(+) T cell repertoire selection occurs early in the immune response in vivo and suggest that persistence and expansion of a population of oligoclonal, intermediate affinity T cells is involved in CD4(+) T cell memory.

    View details for Web of Science ID 000166013100005

    View details for PubMedID 11120769

  • Rapid and efficient vascular transport of arginine polymers inhibits myointimal hyperplasia CIRCULATION Uemura, S., Fathman, C. G., Rothbard, J. B., Cooke, J. P. 2000; 102 (21): 2629-2635

    Abstract

    We recently discovered that short polymers of arginine efficiently translocate across the cytoplasmic membrane independent of the basic amino acid transporter. We evaluated the kinetics and biological effects of heptamers of L-arginine and D-arginine (L-R7 and D-R7, respectively) in vascular cells. We assessed the effects of these peptides on the NO synthesis pathway and vascular cell proliferation.Human umbilical vein endothelial cell and rabbit vascular segments were incubated in medium containing biotin-labeled L-R7 or D-R7. Both polymers rapidly translocated through the vessel wall and into the vascular cells in a dose- and time-dependent fashion. At a dose of 10 micromol/L for 30 minutes, 100% of the endothelial cells showed evidence of cytoplasmic and nuclear localization of the peptides. To evaluate the biological effects of the polymer translocation on myointimal formation, rabbit jugular vein segments were incubated with polymers (10 micromol/L, 30 minutes) or vehicle before arterial interposition grafting. Planimetric measurement 28 days after surgery revealed that L-R7 and D-R7 substantially reduced myointimal formation compared with the control condition (intima/media ratio: control 1. 50.5, L-R7 0.40.2, and D-R7 0.80.2; P:<0.05). Furthermore, basal nitrate and nitrite production from L-R7-treated grafts was significantly higher than that from both control and D-R7-treated veins. Studies in vitro of cultured vascular smooth muscle cells revealed that both polymers also exhibit an NO-independent inhibition of vascular smooth muscle cell proliferation.Short polymers of arginine have the unique ability of vascular cell translocation, and they also have direct biological effects. These attributes are potentially useful in treating myointimal hyperplasia.

    View details for Web of Science ID 000165405800013

    View details for PubMedID 11085967

  • Polyarginine enters cells more efficiently than other polycationic homopolymers JOURNAL OF PEPTIDE RESEARCH Mitchell, D. J., Kim, D. T., Steinman, L., Fathman, C. G., Rothbard, J. B. 2000; 56 (5): 318-325

    Abstract

    Homopolymers or peptides containing a high percentage of cationic amino acids have been shown to have a unique ability to cross the plasma membrane of cells, and consequently have been used to facilitate the uptake of a variety of biopolymers and small molecules. To investigate whether the polycationic character of these molecules, or some other structural feature, was the molecular basis for the effect, the ability of a variety of homopolymers to enter cells was assayed by confocal microscopy and flow cytometry. Polymers of L- or D-arginine containing six or more amino acids entered cells far more effectively than polymers of equal length composed of lysine, ornithine and histidine. Peptides of fewer than six amino acids were ineffective. The length of the arginine side-chain could be varied without significant loss of activity. These data combined with the inability of polymers of citrulline to enter cells demonstrated that the guanidine headgroup of arginine was the critical structural component responsible for the biological activity. Cellular uptake could be inhibited by preincubation of the cells with sodium azide, but not by low temperature (3 degrees C), indicating that the process was energy dependent, but did not involve endocytosis.

    View details for Web of Science ID 000165173900006

    View details for PubMedID 11095185

  • Targeting rare populations of murine antigen-specific T lymphocytes by retroviral transduction for potential application in gene therapy for autoimmune disease JOURNAL OF IMMUNOLOGY Costa, G. L., Benson, J. M., Seroogy, C. M., Achacoso, P., Fathman, C. G., Nolan, G. P. 2000; 164 (7): 3581-3590

    Abstract

    CD4+ T cells are important mediators in the pathogenesis of autoimmunity and would therefore provide ideal candidates for lymphocyte-based gene therapy. However, the number of Ag-specific T cells in any single lesion of autoimmunity may be quite low. Successful gene transfer into autoantigen-specific CD4+ T cells would serve as an ideal vehicle for site-targeted gene therapy if it were possible to transduce preferentially the small number of autoantigen-specific T cells. In this study we have demonstrated that retroviral infection of CD4+ lymphocytes from either autoantigen-stimulated TCR transgenic mice, or Ag-activated immunized nontransgenic mice, with a retroviral vector (pGCIRES), resulted in the transduction of only the limited number of Ag-reactive CD4+ T cells. In contrast, polyclonal activation of the same cultures resulted in transduction of non-antigen-specific lymphocytes. Transduction of Ag-reactive CD4+ T cells with pGCIRES retrovirus encoding the regulatory genes IL-4 (IL4) and soluble TNF receptor (STNFR) resulted in stable integration and long-term expression of recombinant gene products. Moreover, expression of the pGCIRES marker protein, GFP, directly correlated with the expression of the upstream regulatory gene. Retroviral transduction of CD4+ T cells targeted specifically Ag-reactive cells and was cell cycle-dependent and evident only during the mitosis phase. These studies suggest that retroviral transduction of autoantigen-specific murine CD4+ T cells, using the pGCIRES retroviral vector, may provide a potential method to target and isolate the low frequency of autoantigen-specific murine CD4+ T cells, and provides a rational approach to gene therapy in animal models of autoimmunity.

    View details for Web of Science ID 000086020700019

    View details for PubMedID 10725713

  • Identification and characterization of the antigen-specific subpopulation of alloreactive CD4+T cells in vitro and in vivo TRANSPLANTATION Krieger, N. R., Fathman, C. G., Shaw, M. K., Ridgway, W. M. 2000; 69 (4): 605-609

    Abstract

    We report the identification and characterization of the small subpopulation of alloantigen-specific T cells in vitro and in vivo. This subpopulation of T cells was distinguished by up-regulation of cell surface CD4 expression. These CD4high T cells were alloantigen specific in proliferation assays in vitro, and they expressed memory/activation markers, including CD44high and CD69high. Further studies demonstrated that these allospecific CD4high cells were also present (< or = 1% of CD4+ T cells) in vivo in BALB/c (H-2d) recipients of C57BL/6 (H-2b) skin allografts. CD4high T cells isolated from regional draining lymph nodes in these skin graft recipients reacted in a donor-specific fashion to C57BL/6 splenocyte stimulator cells in mixed lymphocyte culture. Adoptive transfer of CD4high, but not CD4normal T cells, just before skin engraftment in CD4 knockout mice, reconstituted rejection. The discovery that a small subpopulation of CD4high lymph node cells contained all of the alloantigen-specific T cells may allow study of tissue-specificity and subsequent alloantigen identification in transplantation.

    View details for Web of Science ID 000085611500023

    View details for PubMedID 10708118

  • Application of gene therapy in autoimmune disease. Current directions in autoimmunity Fathman, C. G., Seroogy, C. M. 2000; 2: 189-202

    View details for PubMedID 11791456

  • The application of gene therapy in autoimmune diseases GENE THERAPY Seroogy, C. M., Fathman, C. G. 2000; 7 (1): 9-13

    Abstract

    The application of gene therapy in autoimmune disease represents a novel use of this technology. The goal of gene therapy in the treatment of autoimmune disease is to restore 'immune homeostasis' by countering the pro-inflammatory effects of the CD4+ T cells in the lesions of autoimmunity. This can be accomplished by adoptive therapy with transduced T cells which can specifically home to the site of inflammation and secrete 'regulatory' protein(s) to ameliorate the inflammation or by direct targeting of the retroviral vector to activated T cells in the sites of inflammation. Transduction of autoantigen recognizing CD4+ T cells, to secrete anti-inflammatory products, may become the 'magic bullet' to combat the ravages of autoimmune inflammation and tissue destruction. Gene Therapy (2000) 7, 9-13.

    View details for Web of Science ID 000084878300002

    View details for PubMedID 10680009

  • Isolation of self antigen-reactive cells from inflamed islets of nonobese diabetic mice using CD4(high) expression as a marker JOURNAL OF IMMUNOLOGY Lejon, K., Fathman, C. G. 1999; 163 (10): 5708-5714

    Abstract

    The low precursor frequency of Ag-reactive CD4+ T cells has been a barrier to the study of CD4+ T cell responses to conventional Ags as well as CD4+ T cell responses to autoantigens recognized during the course of an autoimmune disease. We have recently reported that all "conventional Ag" reactive CD4+ T cells are contained within the subpopulation expressing high levels of the CD4 molecule, termed CD4high. We have identified a CD4high population in the islets of Langerhans of prediabetic nonobese diabetic (NOD) mice that is extremely potent in transferring disease. As few as 500 CD4high islet-infiltrating CD4+ T cells transferred insulin-dependent diabetes mellitus to CD8 reconstituted NOD-SCID mice within 30 days of transfer. In contrast, CD4high T cells isolated from either NOD spleen or salivary glands did not transfer insulin-dependent diabetes mellitus into similar CD8-reconstituted NOD-SCID recipients. These data indicate that the precursor frequency of NOD islet-reactive, pathogenic CD4+ T cells is much higher in the prediabetic NOD pancreas than in these other organs. The islet-infiltrating CD4high T cells displayed selected memory markers, by cell surface analysis, and displayed a Th 1 phenotype by RNase protection assay, but had a marked decrease in IL-4 mRNA determined by quantitative real time PCR when compared with the less pathogenic CD4normal islet-infiltrating T cells. Use of the CD4high marker to select Ag activated T cells represents a tool to isolate and study pathogenic CD4+ T cells from autoimmune lesions in which the Ag has not been previously defined.

    View details for Web of Science ID 000083638400067

    View details for PubMedID 10553102

  • A new look at MHC and autoimmune disease. Science Ridgway, W. M., Fassò, M., Fathman, C. G. 1999; 284 (5415): 749-?

    View details for PubMedID 10336398

  • Suppressive immunization with DNA encoding a self-peptide prevents autoimmune disease: Modulation of T cell costimulation JOURNAL OF IMMUNOLOGY Ruiz, P. J., Garren, H., Ruiz, I. U., Hirschberg, D. L., Nguyen, L. V., Karpuj, M. V., Cooper, M. T., Mitchell, D. J., Fathman, C. G., Steinman, L. 1999; 162 (6): 3336-3341

    Abstract

    Usually we rely on vaccination to promote an immune response to a pathogenic microbe. In this study, we demonstrate a suppressive from of vaccination, with DNA encoding a minigene for residues 139-151 of myelin proteolipid protein (PLP139-151), a pathogenic self-Ag. This suppressive vaccination attenuates a prototypic autoimmune disease, experimental autoimmune encephalomyelitis, which presents clinically with paralysis. Proliferative responses and production of the Th1 cytokines, IL-2 and IFN-gamma, were reduced in T cells responsive to PLP139-151. In the brains of mice that were successfully vaccinated, mRNA for IL-2, IL-15, and IFN-gamma were reduced. A mechanism underlying the reduction in severity and incidence of paralytic autoimmune disease and the reduction in Th1 cytokines involves altered costimulation of T cells; loading of APCs with DNA encoding PLP139-151 reduced the capacity of a T cell line reactive to PLP139-151 to proliferate even in the presence of exogenous CD28 costimulation. DNA immunization with the myelin minigene for PLP-altered expression of B7.1 (CD80), and B7.2 (CD86) on APCs in the spleen. Suppressive immunization against self-Ags encoded by DNA may be exploited to treat autoimmune diseases.

    View details for Web of Science ID 000079105000029

    View details for PubMedID 10092787

  • Local delivery of TNF by retrovirus-transduced T lymphocytes exacerbates experimental autoimmune encephalomyelitis CLINICAL IMMUNOLOGY Dal Canto, R. A., Shaw, M. K., Nolan, G. P., Steinman, L., Fathman, C. G. 1999; 90 (1): 10-14

    Abstract

    Experimental autoimmune encephalomyelitis (EAE) is an inflammatory autoimmune disease of the central nervous system that serves as a model for the human disease multiple sclerosis. Paralysis is "induced" by CD4+ T cells of the Th1 phenotype. Tumor necrosis factor (TNF), a Th1 type cytokine, has been shown to be upregulated in the CNS during the onset of EAE, and systemic manipulations of TNF have had substantial effects on disease progression. However, the precise role of TNF in EAE has been called into question by recent experiments utilizing TNF and lymphotoxin knockout mice. We demonstrate here that the local delivery of TNF by myelin basic protein (MBP)-specific T cells, retrovirally transduced to express TNF, exacerbated MBP-induced disease following adoptive transfer into syngeneic mice.

    View details for Web of Science ID 000079158600002

    View details for PubMedID 9884347

  • Analysis of the role of variation of major histocompatibility complex class II expression on nonobese diabetic (NOD) peripheral T cell response JOURNAL OF EXPERIMENTAL MEDICINE Ridgway, W. M., Ito, H., Fasso, M., Yu, C., Fathman, C. G. 1998; 188 (12): 2267-2275

    Abstract

    The current paradigm of major histocompatibility complex (MHC) and disease association suggests that efficient binding of autoantigens by disease-associated MHC molecules leads to a T cell-mediated immune response and resultant autoimmune sequelae. The data presented below offer a different model for this association of MHC with autoimmune diabetes. We used several mouse lines expressing different levels of I-Ag7 and I-Ak on the nonobese diabetic (NOD) background to evaluate the role of MHC class II in the previously described NOD T cell autoproliferation. The ratio of I-Ag7 to I-Ak expression correlated with the peripheral T cell autoproliferative phenotype in the mice studied. T cells from the NOD, [NOD x NOD. I-Anull]F1, and NOD I-Ak transgenic mice demonstrated autoproliferative responses (after priming with self-peptides), whereas the NOD.H2(h4) (containing I-Ak) congenic and [NOD x NOD. H2(h4) congenic]F1 mice did not. Analysis of CD4(+) NOD I-Ak transgenic primed lymph node cells showed that autoreactive CD4(+) T cells in the NOD I-Ak transgenic mice were restricted exclusively by I-Ag7. Considered in the context of the avidity theory of T cell activation and selection, the reported poor peptide binding capacity of NOD I-Ag7 suggested a new hypothesis to explain the effects of MHC class II expression on the peripheral autoimmune repertoire in NOD mice. This new explanation suggests that the association of MHC with diabetes results from "altered" thymic selection in which high affinity self-reactive (potentially autoreactive) T cells escape negative selection. This model offers an explanation for the requirement of homozygous MHC class II expression in NOD mice (and in humans) in susceptibility to insulin-dependent diabetes mellitus.

    View details for Web of Science ID 000077713600008

    View details for PubMedID 9858513

  • T cell receptor (TCR) engagement leads to activation-induced splicing of tumor necrosis factor (TNF) nuclear pre-mRNA JOURNAL OF EXPERIMENTAL MEDICINE Yang, Y., Chang, J. F., Parnes, J. R., Fathman, C. G. 1998; 188 (2): 247-254

    Abstract

    Inducible gene expression is primarily regulated at the level of transcription. Additional steps of "processing" pre-mRNA, involved in the regulation of induced gene expression, have not been previously reported. Here we report a novel mechanism of "activation-induced splicing" of preexisting tumor necrosis factor (TNF) message (pre-mRNA) in naive T lymphocytes after engagement of the T cell receptor (TCR), which still occurs after inhibition of transcription. Expression of TNF has been previously demonstrated to be regulated at both the transcriptional and translational levels. However, neither the large pool of TNF mRNA observed in activated T cells nor TNF protein production, which peaks very shortly after activation, can be solely attributed to increased transcription. Evidence is presented that activation-induced splicing of TNF pre-mRNA plays a significant role in the rapid production of TNF seen in activated T cells. Activation triggers processing of TNF pre-mRNA that has accumulated in naive T cells (before activation-induced transcription), and the mature TNF mRNA is translocated to the cytoplasm for rapid translation and protein production. This novel form of activation-induced splicing of TNF may allow T cells to mount an immediate response to activation stimuli under physiological conditions.

    View details for Web of Science ID 000075236900003

    View details for PubMedID 9670037

  • Following antigen challenge, T cells up-regulate cell surface expression of CD4 in vitro and in vivo JOURNAL OF IMMUNOLOGY Ridgway, W., Fasso, M., Fathman, C. G. 1998; 161 (2): 714-720

    Abstract

    The low precursor frequency of Ag-specific T cells has raised significant barriers to studying the T cell response in vivo. We demonstrate that T cells up-regulate the cell surface expression of CD4 following Ag recognition, which identifies Ag-specific T cells in vitro and in vivo and allows their characterization. The CD4high cell subpopulation contains the Ag-specific population as indicated by Ag-induced proliferation and limiting dilution analyses. The use of the CD4high marker will allow analysis of the dynamics of the T cell immune response in vivo, the study of the suboptimal T cell response to Ag, and the identification of T cells which are reactive to known and unknown autoantigens.

    View details for Web of Science ID 000074728400024

    View details for PubMedID 9670947

  • Prolongation of cardiac graft survival with anti-CD4Ig plus hCTLA4Ig in primates JOURNAL OF SURGICAL RESEARCH Krieger, N. R., Yuh, D., McIntyre, W. B., Flavin, T. F., Yin, D. P., Robbins, R., Fathman, C. G. 1998; 76 (2): 174-178

    Abstract

    The aim of this study was to determine whether the use of combined immunotherapy with a brief course of humanized anti-CD4Ig and hCTLA4Ig would prolong heterotopic cardiac allograft survival in primates (rhesus monkeys). This model was based on work in "high responder" rats where a brief course of depletive anti-CD4mAb plus hCTLA4Ig was successful in inducing transplantation tolerance.Heterotopic cardiac transplants were performed in rhesus recipients. Donor/recipient pairs between groups were confirmed to be reactive prior to transplantation by MLR matching. Humanized anti-CD4Ig, a recently developed anti-CD4mAb, was given at a dose of 20 mg/kg i.v. on days -3, -2, -1, and 0. hCTLA4Ig was administered at 6 mg/kg/dose i.v. on days 0 and 2 for the first recipient and days 0, 2, 4, and 6 for the second recipient. No further immunosuppression was administered. The treated (n = 2) or untreated (n = 5) recipients were followed for graft function by daily palpitation.Treatment with anti-CD4Ig plus hCTLA4Ig resulted in a significant prolongation of heart graft survival (42 days for the first recipient and 52 days for the second recipient) compared to untreated recipients (7 days x 4, 11 days x 1). FACS analysis demonstrated CD4 depletion of anti-CD4 treated animals to <2% on posttransplant day 1. The CD4+ T cells gradually repopulated to 50-70% pretransplant levels just prior to rejection. No adverse responses (fever, tachypnea, tachycardia, infections) were observed.These are the first results demonstrating that a brief course of combined specific induction immunotherapy with humanized anti-CD4Ig plus hCTLA4Ig, in the absence of adjuvant posttransplant immunosuppression, was well tolerated and resulted in marked prolongation of cardiac allograft survival in primates.

    View details for Web of Science ID 000075343200012

    View details for PubMedID 9698519

  • Regulation of programmed cell death following T cell activation in vivo INTERNATIONAL IMMUNOLOGY Yang, Y., Kim, D., Fathman, C. G. 1998; 10 (2): 175-183

    Abstract

    Activation of T cell hybridomas in vitro induces rapid Fas-Fas ligand (FasL)-mediated programmed cell death (apoptosis). In contrast, T cells activated by antigen or superantigen in vivo undergo a population expansion and then decline due to Fas-FasL-mediated activation-induced apoptosis (AIA). We asked how T cells activated by antigen in vivo proliferated before undergoing apoptosis. Two possibilities were analyzed: either (i) the apoptosis program was not 'turned on' or (ii) was 'blocked' during the period of cellular proliferation in vivo. Data presented in this manuscript support the second of these possibilities. CD4+ T cells activated in vivo were resistant to anti-fas-mediated apoptosis until 48 h following staphylococcal enterotoxin B (SEB) administration, despite the fact that activated proliferating T cells expressed high levels of Fas (CD95) antigen and many 'apoptosis genes' were induced within 24 h of SEB administration. The analysis of the expression patterns of 'apoptosis genes' during the T cell activation further suggested that temporal blockade of AIA may be due to the induction of apoptosis-preventing genes, such as bag-1.

    View details for Web of Science ID 000072440200009

    View details for PubMedID 9533445

  • A gene therapy approach to treatment of autoimmune disease IMMUNOLOGIC RESEARCH Seroogy, C. M., Fathman, C. G. 1998; 18 (1): 15-26

    Abstract

    New insights into the underlying mechanisms for the development of autoimmune diseases in humans and various animal models continue to increase with our understanding of factors that drive polarization of T helper (Th) responses and tolerance. This information has led to the development of new treatment strategies, including oral tolerance clinical trails and the use of altered peptide ligands in animal models. These approaches have shown some promise and provided additional insight into the disease processes. The use of gene therapy in many disease states continues to increase. We are starting to see the application of gene therapy in chronic diseases in humans. Gene therapy has been used in several animal models of autoimmune disease with promising preliminary results. In this article, an overview will be provided for the use of gene therapy in autoimmune disease.

    View details for Web of Science ID 000075425800002

    View details for PubMedID 9724846

  • The association of MHC with autoimmune diseases: Understanding the pathogenesis of autoimmune diabetes CLINICAL IMMUNOLOGY AND IMMUNOPATHOLOGY Ridgway, W. M., Fathman, C. G. 1998; 86 (1): 3-10

    Abstract

    The current paradigm of MHC and disease association is efficient binding of autoantigens by disease-associated MHC molecules leading to a T cell-mediated immune response and resultant autoimmune sequelae. Data presented here offer a different model for this association of MHC with autoimmune diabetes. This new explanation suggests that the association of MHC with autoimmunity results from "altered" thymic selection in which high-affinity self-reactive (potentially autoreactive) T cells escape negative selection. This model offers an explanation for the requirement of homozygous MHC class II expression in NOD mice (and in man) in susceptibility to IDDM.

    View details for Web of Science ID 000071367200002

    View details for PubMedID 9434791

  • CD45RB(high) CD4(+) T cells from IFN-gamma knockout mice do not induce wasting disease JOURNAL OF AUTOIMMUNITY Ito, H., Fathman, C. G. 1997; 10 (5): 455-459

    Abstract

    Transfer of CD45RBhigh CD4+ T cells from normal mice to congenic SCID mice induces wasting disease, a murine model of inflammatory bowel disease. In this model, colonic inflammation is considered to be caused by a disregulated Th1 response, and Th1 cytokines, especially IFN-gamma, have been suggested to play an important role in the pathogenesis of wasting disease. In order to elucidate the potential role of IFN-gamma in the pathogenesis of wasting disease, we transferred CD45RBhigh CD4+ T cells from IFN-gamma knockout (GKO) mice to congenic SCID mice. The recipient mice were absolutely free of symptoms and clinical signs of disease and showed body-weight gain similar to that seen in normal mice. These data demonstrate the essential and non-redundant role of IFN-gamma in the pathogenesis of wasting disease.

    View details for Web of Science ID A1997YA16000005

    View details for PubMedID 9376073

  • Introduction of soluble proteins into the MHC class I pathway by conjugation to an HIV tat peptide JOURNAL OF IMMUNOLOGY Kim, D. T., Mitchell, D. J., Brockstedt, D. G., Fong, L., Nolan, G. P., Fathman, C. G., Engleman, E. G., Rothbard, J. B. 1997; 159 (4): 1666-1668

    Abstract

    Protection against most intracellular pathogens requires T cells that recognize pathogen-derived peptides in association with MHC class I molecules on the surface of infected cells. However, because exogenous proteins do not ordinarily enter the cytosol and access the MHC class I-processing pathway, protein-based vaccines that induce class I-restricted CTL responses have proved difficult to design. We have addressed this problem by conjugating proteins, such as OVA, to a short cationic peptide derived from HIV-1 tat (residues 49-57). When APC were exposed in vitro to such protein conjugates, they processed and presented the peptides in association with MHC class I molecules and stimulated CD8+ Ag-specific T cells. Moreover, Ag-specific CTLs were generated in vivo by immunizing mice with histocompatible dendritic cells that had been exposed to protein-tat conjugates.

    View details for Web of Science ID A1997XP43800012

    View details for PubMedID 9257826

  • Local delivery of interleukin 4 by retrovirus-transduced T lymphocytes ameliorates experimental autoimmune encephalomyelitis JOURNAL OF EXPERIMENTAL MEDICINE Shaw, M. K., Lorens, J. B., Dhawan, A., DalCanto, R., Tse, H. Y., Tran, A. B., Bonpane, C., Eswaran, S. L., Brocke, S., Sarvetnick, N., Steinman, L., Nolan, G. P., Fathman, C. G. 1997; 185 (9): 1711-1714

    Abstract

    Experimental autoimmune encephalomyelitis (EAE) is an inflammatory autoimmune disease of the central nervous system which serves as a model for the human disease multiple sclerosis. We demonstrate here that encephalitogenic T cells, transduced with a retroviral gene, construct to express interleukin 4, and can delay the onset and reduce the severity of EAE when adoptively transferred to myelin basic protein-immunized mice. Thus, T lymphocytes transduced with retroviral vectors can deliver "regulatory cytokines" in a site-specific manner and may represent a viable therapeutic strategy for the treatment of autoimmune disease.

    View details for Web of Science ID A1997WY11700020

    View details for PubMedID 9151908

  • The use of CD4 and CD8 knockout mice to study the role of T-cell subsets in allotransplant rejection JOURNAL OF HEART AND LUNG TRANSPLANTATION Krieger, N. R., Fathman, C. G. 1997; 16 (3): 263-267

    View details for Web of Science ID A1997WQ62800001

    View details for PubMedID 9087868

  • Induction of tolerance to small bowel allografts in high-responder rats by combining anti-CD4 with CTLA4Ig TRANSPLANTATION Yin, D. P., Sankary, H. N., Williams, J., Krieger, N., Fathman, C. G. 1996; 62 (11): 1537-1539

    Abstract

    This study was designed to investigate the effectiveness of combined perioperative anti-CD4 and human (h)CTLA4Ig therapy in preventing allorejection of small bowel transplantation in high-responder Lewis rat recipients of ACI grafts. Anti-CD4 (5 mg/kg x 4 days) or hCTLA4Ig (0.5 mg/rat x 2 days) therapy alone delayed, but did not prevent, allograft rejection after small bowel transplantation of ACI into Lewis rats. All grafts were rejected in 18 and 10 days, respectively. However, a regimen of anti-CD4 (5 mg/kg x 4 days) combined with hCTLA4Ig (0.5 mg/rat x 2 days) allowed indefinite survival of ACI small bowel allografts. Second donor-matched heart grafts were permanently accepted, whereas third-party (Sprague-Dawley) heart allografts were rejected by the tolerant recipients. These data suggest that these two reagents produced a synergistic effect in preventing allorejection of small bowel transplantation.

    View details for Web of Science ID A1996VZ23500001

    View details for PubMedID 8970603

  • CD4(+) but not CD8(+) cells are essential for allorejection JOURNAL OF EXPERIMENTAL MEDICINE Krieger, N. R., Yin, D. P., Fathman, C. G. 1996; 184 (5): 2013-2018

    Abstract

    The generation of knockout mice with targeted gene disruption has provided a valuable tool for studying the immune response. Here we describe the use of CD4 and CD8 knockout mice to examine the role of CD4+ and CD8+ cells in initiating allotransplantation rejection. Pretreatment with a brief course of depletive anti-CD4 monoclonal antibody therapy allowed permanent survival of heart, but not skin, allografts transplanted across a major histocompatibility barrier. However, skin as well as heart grafts were permanently accepted in the CD4 knockout mice. Transfer of CD4+ cells into CD4 knockout recipient mice 1 d before skin engraftment reconstituted rejection, demonstrating that CD4+ cells are necessary for initiating rejection of allogeneic transplants. Major histocompatibility complex disparate heart and skin allografts transplanted into CD8 knockout recipients were rejected within 10 d. This study demonstrates that CD4+ but not CD8+ T cells are absolutely required to initiate allograft rejection.

    View details for Web of Science ID A1996VT69400040

    View details for PubMedID 8920888

  • beta-cell destruction may be a late consequence of the autoimmune process in nonobese diabetic mice DIABETES Shimada, A., Charlton, B., TAYLOREDWARDS, C., Fathman, C. G. 1996; 45 (8): 1063-1067

    Abstract

    The NOD mouse is an animal model of IDDM that shows many of the characteristics of human IDDM. It has been proposed that beta-cell destruction in IDDM progresses over time in a linear manner. Recently, we and others have demonstrated that T helper type 1 (Th1) cells have pathogenic roles in the NOD model and proposed that cytokine balances change as the disease progresses. However, it has not been demonstrated how or when the cytokine balances change or how the beta-cell destruction progresses. We have recently demonstrated that the cytokine profiles of CD45RB(low) CD4+ cells correlate either with their pathogenic or with their protective roles in the NOD mouse. To further analyze this apparent correlation between the shift in cytokine level and IDDM, we examined the anti-CD3-induced cytokine profiles of this subset from NOD mice of various ages compared with that from age-matched I-Ak transgenic NOD and BALB/c mice as controls. A significantly higher ratio of anti-CD3-induced interferon-gamma/interleukin-4 was found in diabetic NOD mice (P < 0.0001) but not in age-matched nondiabetic NOD mice. This cytokine ratio did not change significantly until the onset of diabetes in NOD mice. Based upon these results, we propose that IDDM in the NOD mouse progresses as a predominant inflammatory beta-cell dysfunction without actual beta-cell destruction until late in the disease process. This supports the possibility that late-stage immunotherapy may preserve islet beta-cell mass.

    View details for Web of Science ID A1996VA07300010

    View details for PubMedID 8690153

  • Quantitative analysis of T cell activation - Role of TCR ligand density and TCR affinity JOURNAL OF IMMUNOLOGY Kim, D. T., Rothbard, J. B., Bloom, D. D., Fathman, C. G. 1996; 156 (8): 2737-2742

    Abstract

    (B6 X A)F1 mice were immunized with sperm whale myoglobin, and T cell clones and hybridomas were generated. Hybridoma 74a.e9 was specific for the sperm whale myoglobin 67-79 peptide and could be partially activated by a peptide analogue, equine myoglobin with a natural 74G substitution. Using this hybridoma in T cell activation assays, we studied the effects of varying the avidity of the TCR for its ligand, the concentration of MHC:peptide complex on the APC, and the density of TCR on the surface. Varying ligand concentration on the surface of the APC, the TCR avidity, or the density of TCR on the T cell were equally important parameters in driving T cell activation. The mouse myoglobin (74T) analogue, however, acted as an antagonist to the T cell response. Its effectiveness was also partially determined by its ability to bind to MHC. By independently altering each of these variables and following T cell activation, we describe the interrelationships among these three components (MHC:peptide:TCR) that control the activation of the T cell.

    View details for Web of Science ID A1996UE19200013

    View details for PubMedID 8609391

  • Breaking self-tolerance in nonobese diabetic mice JOURNAL OF EXPERIMENTAL MEDICINE Ridgway, W. M., Fasso, M., LANCTOT, A., Garvey, C., Fathman, C. G. 1996; 183 (4): 1657-1662

    Abstract

    Unresponsiveness to self is maintained through two mechanisms of immune regulation: thymic-negative selection and peripheral tolerance. Although thymic-negative selection is a major mechanism to eliminate self-reactive T cells, normal mice have readily detectable populations of T cells reactive to self-proteins but do not exhibit autoimmune responses. It has been postulated that autoimmune disease results from breakdown or loss of peripheral tolerance. We present data that demonstrate that peripheral tolerance or unresponsiveness to self can be broken in nonobese diabetic (NOD) mice. Immunization of NOD mice (but not of conventional mice) with self-peptides caused an immune response to self-peptide with resultant autoproliferation of peripheral lymphocytes. Autoproliferation of self-reactive T cells in NOD mice resulted from the recognition and proliferation of the activated T cells to endogenously processed and presented self-antigens. This loss of self-tolerance demonstrated in vitro may well be the basis of NOD autoimmune disease in vivo.

    View details for Web of Science ID A1996UH14400039

    View details for PubMedID 8666923

  • Monoclonal T cells identified in early NOD islet infiltrates IMMUNITY Yang, Y., Charlton, B., Shimada, A., DalCanto, R., Fathman, C. G. 1996; 4 (2): 189-194

    Abstract

    To examine the hypothesis that a single initiating antigen was recognized by a monoclonal T cell population leading to subsequent inflammatory insulitis in non-obese (NOD) mouse islets, we examined the T cell receptor TCR V beta repertoire of islet-infiltrating T cells in very young (2-week-old) NOD mice. In independent experiments, we repeatedly identified one monoclonal TCR V-beta 8.2 gene product expressed by T lymphocytes infiltrating the islets of NOD mice at 2 weeks of age. The resultant inflammatory response quickly obscures the monoclonal nature of the initiating event. These data suggest that autoimmune diabetes in NOD mice may be initiated by recognition of a single autoantigen.

    View details for Web of Science ID A1996TX22500009

    View details for PubMedID 8624809

  • Treatment of experimental encephalomyelitis with a peptide analogue of myelin basic protein NATURE Brocke, S., Gijbels, K., Allegretta, M., Ferber, I., Piercy, C., Blankenstein, T., Martin, R., Utz, U., Karin, N., Mitchell, D., VEROMAA, T., Waisman, A., Gaur, A., Conlon, P., Ling, N., Fairchild, P. J., Wraith, D. C., OGARRA, A., Fathman, C. G., Steinman, L. 1996; 379 (6563): 343-346

    Abstract

    Following induction of experimental encephalomyelitis with a T-cell clone, L10C1, that is specific for the myelin basic protein epitope p87-99, the inflammatory infiltrate in the central nervous system contains a diverse collection of T cells with heterogeneous receptors. We show here that when clone L10C1 is tolerized in vivo with an analogue of p87-99, established paralysis is reversed, inflammatory infiltrates regress, and the heterogeneous T-cell infiltrate disappears from the brain, with only the T-cell clones that incited disease remaining in the original lesions. We found that antibody raised against interleukin-4 reversed the tolerance induced by the altered peptide ligand. Treatment with this altered peptide ligand selectively silences pathogenic T cells and actively signals for the efflux of other T cells recruited to the site of disease as a result of the production of interleukin-4 and the reduction of tumour-necrosis factor-alpha in the lesion.

    View details for Web of Science ID A1996TR32300058

    View details for PubMedID 8552189

  • Pathogenic and protective roles of CD45RB(low) CD4(+) cells correlate with cytokine profiles in the spontaneously autoimmune diabetic mouse DIABETES Shimada, A., Rohane, P., Fathman, C. G., Charlton, B. 1996; 45 (1): 71-78

    Abstract

    The adoptive transfer of splenocytes from diabetic NOD mice to NOD-scid/scid (NOD-scid) recipients results in diabetes. This model was used to test the effect of cotransfer of splenocyte subsets from young nondiabetic NOD mice. As shown previously in other NOD models, the CD4+ subset from young nondiabetic mice significantly delayed the onset of diabetes in splenocyte cotransfers (P < 0.001). The data presented here showed that the development of diabetes in NOD-scid recipients correlated with a rapid increase in peripheral CD45RB(low) CD4+ cells. However, the CD45RB(low) subset of CD4+ cells from young nondiabetic mice protected from diabetes transfer in this model. We therefore examined whether CD45RB(low) CD4+ cells from diabetic mice were pathogenic rather than protective. CD45RB(low) CD4+ splenocytes from diabetic NOD mice were transferred along with CD8+ splenocytes from diabetic mice into NOD-scid recipients, and all of the recipients became diabetic within 5 weeks posttransfer. In contrast, no recipients (0 of 10) of CD45RB(high) CD4+ cells along with CD8+ splenocytes from diabetic mice became diabetic within 5 weeks posttransfer (P < 0.001). A correlate for the difference between CD45RB(low) CD4+ cells from diabetic NOD mice and CD45RB(low) CD4+ cells from nondiabetic mice, which showed protective effect in splenocyte cotransfers, was found in cytokine production after stimulation with anti-CD3 antibodies in vitro. CD45RB(low) CD4+ cells from diabetic mice showed a significantly higher ratio (approximately fivefold) of gamma-interferon (IFN-gamma) to interleukin (IL)-4 when compared with CD45RB(low) CD4+ cells from nondiabetic mice (P < 0.001). In conclusion, the function of the CD45RB(low) population of CD4+ cells changes from a protective to a pathogenic one during the development of disease in the NOD mouse. This change in function correlates with cytokine production in vitro; increased IFN-gamma-to-IL-4 ratio is associated with pathogenic potential and occurs coincident with (or after) the onset of diabetes.

    View details for Web of Science ID A1996TM67700011

    View details for PubMedID 8522063

  • Mice with a disrupted IFN-gamma gene are susceptible to the induction of experimental autoimmune encephalolmyelitis (EAE) JOURNAL OF IMMUNOLOGY Ferber, I. A., Brocke, S., TAYLOREDWARDS, C., Ridgway, W., Dinisco, C., Steinman, L., Dalton, D., Fathman, C. G. 1996; 156 (1): 5-7

    Abstract

    Experimental autoimmune encephalomyelitis (EAE), an animal model for multiple sclerosis, is an autoimmune disorder seen in mice and rats following immunization with myelin basic protein (MBP) or MBP-derived peptides. IFN-gamma, a cytokine produced by a variety of cells, is involved in many inflammatory and immune regulatory events. Contradictory results concerning exacerbations and the disease course were seen comparing injections of IFN-gamma in humans suffering from multiple sclerosis to studies using anti-IFN-gamma Abs in mice with EAE. To study the role of IFN-gamma and IFN-gamma-producing cells in EAE, we crossed IFN-gamma knockout mice (H-2b) (unable to produce IFN-gamma due to the disruption of the IFN-gamma gene) with an EAE-susceptible mouse strain, B10.PL (H-2u). EAE was seen in IFN-gamma knockout mice, heterozygotic (IFN-gamma +/-) mice, as well as wild-type littermates following immunization with MBP. Histologic analyses of the central nervous system of IFN-gamma knockout mice with EAE revealed massive infiltrates composed of lymphocytes, macrophages, and granulocytes. We conclude that the presence of IFN-gamma is not crucial to the induction or the clinical course of EAE.

    View details for Web of Science ID A1996TM17100002

    View details for PubMedID 8598493

  • Tissue-specific effects of anti-CD4 therapy in induction of allograft unresponsiveness in high and low responder rats. Transplant immunology Yin, D., Fathman, C. G. 1995; 3 (3): 258-264

    Abstract

    In these experiments, we studied the role of anti-CD4 (Ox38) monoclonal antibody in the prevention of heart and/or kidney allograft rejection in low (ACI) and high (Lewis) responder rats. In low responder ACI rats, donor-specific tolerance for heart and kidney allografts (individually or in combination) was achieved by pretransplant anti-CD4 therapy. In high responder Lewis rats, anti-CD4 therapy alone (or combined with anti-CD8 (Ox8), thymectomy or total lymphoid irradiation) did not prevent first-set rejection of heart allografts. This difference was correlated with a more profound and longer lasting CD4+ cell depletion in the low responder strain. Anti-CD4 treatment, however, produced tolerance of kidney transplants in high responder rats. Additionally, anti-CD4 treatment induced tolerance to heart (as well as kidney) allografts in Lewis recipients of combined kidney and heart allografts from ACI. The effects of anti-CD4 treatment thus depend upon the recipient responder status as well as the organs transplanted and the order of transplantation.

    View details for PubMedID 8581415

  • TERNARY COMPLEX THERAPY FOR AUTOIMMUNE-DISEASE HOSPITAL PRACTICE Fathman, C. G. 1995; 30 (8): 57-?

    Abstract

    Induction of an immune response is remarkably simple, involving the interaction of three components: processed antigen, an MHC class II molecule, and a CD4 T-cell receptor. Each part of this ternary complex represents a novel target for interventions that may block subsequent effector phenomena of autoimmune disease. Such specific treatments will be less toxic than those currently available.

    View details for Web of Science ID A1995RN91300018

    View details for PubMedID 7635912

  • INDUCTION OF TOLERANCE TO HEART ALLOGRAFTS IN HIGH RESPONDER RATS BY COMBINING ANTI-CD4 WITH CTLA4IG JOURNAL OF IMMUNOLOGY Yin, D. P., Fathman, C. G. 1995; 155 (4): 1655-1659

    Abstract

    It has been difficult to induce donor-specific transplantation tolerance in high responder Lewis rats. Results presented below demonstrate that amounts of pretransplant anti-CD4 sufficient to allow allograft tolerance in low responder strains (5 mg/kg x 4 days) did not prevent the acute rejection of ACI heart allografts in high responder Lewis recipients. Higher doses of pretransplant anti-CD4 (10 mg/kg, 15 mg/kg, and 20 mg/kg) given alone could delay but not prevent allograft rejection. Pretransplant anti-CD4 combined with anti-CD8, thymectomy, and total lymphoid irradiation all failed to produce tolerance to ACI heart allografts. However, a regimen of anti-CD4 combined with CTLA4Ig allowed indefinite survival of ACI heart allografts (mean survival time, > 100 day). Second-donor matched heart grafts were permanently accepted, and third-party heart grafts were permanently accepted, and third-party heart allografts were rejected by the tolerant recipients. These results suggest a new combination therapeutic strategy for clinical transplantation.

    View details for Web of Science ID A1995RN46400003

    View details for PubMedID 7636225

  • CD4-POSITIVE SUPPRESSOR CELLS BLOCK ALLOTRANSPLANT REJECTION JOURNAL OF IMMUNOLOGY Yin, D. P., Fathman, C. G. 1995; 154 (12): 6339-6345

    Abstract

    Mechanisms of maintenance of transplantation tolerance induced in adult rats by depletive regimens of anti-CD4 before transplantation of vascularized heart allografts were studied. Despite the fact that there has been little evidence that tolerant lymphocytes could prevent allograft rejection after adoptive transfer, we demonstrated a suppressive role for lymphocytes from tolerant animals in vivo. These experiments analyzed the ability of lymphocytes from tolerant rats to protect passenger leukocyte-depleted Lewis heart grafts that had been "parked" in ACI rats (treated with pretransplant anti-CD4 and maintained for > 100 days) compared with their ability to protect transplantation of fresh Lewis heart grafts in naive ACI rats. Although parked Lewis heart grafts were rejected in unmanipulated ACI recipients, parked hearts (but not naive Lewis heart allografts), were permanently accepted by naive ACI rats when syngeneic tolerant spleen cells were adoptively transferred at the time of transplantation. Further, we demonstrated that the suppressor cells in the tolerant spleen cells were CD4+. These results suggest that CD4+ spleen cells from tolerant rats inhibit allograft recognition and may maintain allograft tolerance by blocking the indirect pathway of allorecognition.

    View details for Web of Science ID A1995RC61000014

    View details for PubMedID 7759872

  • ISLET-INFILTRATING LYMPHOCYTES FROM PREDIABETIC NOD MICE RAPIDLY TRANSFER DIABETES TO NOD-SCID/SCID MICE DIABETES Rohane, P. W., Shimada, A., Kim, D. T., Edwards, C. T., Charlton, B., Shultz, L. D., Fathman, C. G. 1995; 44 (5): 550-554

    Abstract

    In an effort to study the development of diabetes in NOD mice, our laboratory developed a novel adoptive transfer model using NOD-scid/scid (NOD-scid) mice as recipients of islet-infiltrating lymphocytes from donor prediabetic female NOD mice. We first confirmed previous results that demonstrated that splenocytes of diabetic and prediabetic female NOD mice could transfer diabetes to NOD-scid mice. We demonstrated that the kinetics of disease transfer were dependent on the age of transferred lymphocytes and reiterated the kinetics of diabetes in conventional female NOD mice. We then demonstrated that islet-infiltrating lymphocytes from prediabetic female NOD mice could transfer diabetes. In contrast with the age-dependent transfer of diabetes seen using splenocytes, islet-infiltrating lymphocytes obtained from prediabetic female NOD mice aged > or = 40 days rapidly transferred diabetes to NOD-scid recipients. The time required to transfer insulin-dependent diabetes mellitus (IDDM) using islet-infiltrating lymphocytes from young prediabetic mice (25 +/- 9 days) was not statistically different from the time required to transfer IDDM using splenocytes from overtly diabetic mice (32 +/- 5 days). Cotransfer of splenocyte cells or CD4+, but not CD8+ spleen cells, from 60- to 80-day-old prediabetic female NOD mice together with either splenocytes from diabetic mice or islet-infiltrating lymphocytes from prediabetic NOD mice delayed the rapid transfer of IDDM, suggesting that CD4+ cells mediated immunoregulation. Use of the NOD-scid islet-infiltrating lymphocyte-adoptive transfer model should help elucidate the pathophysiology of the early inflammatory events leading to insulitis and subsequent beta-cell destruction.(ABSTRACT TRUNCATED AT 250 WORDS)

    View details for Web of Science ID A1995QZ82000012

    View details for PubMedID 7729614

  • REGULATION OF AUTOIMMUNE-RESPONSE CURRENT OPINION IN IMMUNOLOGY Ridgway, W. M., Weiner, H. L., Fathman, C. G. 1994; 6 (6): 946-955

    Abstract

    Recent work on such apparently disparate fields as T-cell receptor peptide-induced regulation, superantigens, antigen-induced tolerance, models of peripheral tolerance, apoptosis, and T-cell receptor antagonists demonstrates a similarity in immune response from a regulatory perspective. In many systems, a 'tolerance' pathway is observed, characterized broadly as an initial disturbance in the immune system, with a resulting predominance of effector cells, followed by a homeostatic response (often requiring CD8+ cells) which leads the effector population into T-cell receptor downregulation, T-cell inactivation, anergy and, often, eventual apoptotic death. In the regulated immune response, mixed populations of anergized and apoptosing T cells can be found. In some cases, anergy appears to lead to death while, in other instances, cells revert to a functional state. This review focuses on recent papers examining each of these topics in an attempt to obtain a preliminary integrated picture of immune regulation in autoimmune diseases.

    View details for Web of Science ID A1994PZ42600018

    View details for PubMedID 7710719

  • PREVENTION OF DIABETES AND INSULITIS BY NEONATAL INTRATHYMIC ISLET ADMINISTRATION IN NOD MICE JOURNAL OF AUTOIMMUNITY Charlton, B., TAYLOREDWARDS, C., Tisch, R., Fathman, C. G. 1994; 7 (5): 549-560

    Abstract

    The murine model of human insulin dependent diabetes mellitus (IDDM), the non-obese diabetic (NOD) mouse, develops a T cell-dependent destruction of pancreatic islets. While the target antigens are unknown, there is clearly a lack of tolerance to them. Neonatal intrathymic (i.t.) antigen injection has been successfully employed to prevent insulitis in BB rats but previous i.t. islet antigen studies in NOD mice were done on older mice. We have injected syngeneic islets into the thymus of NOD mice at birth and found that diabetes and insulitis can be completely prevented by this procedure. The effect is islet antigen-specific since other T cell responses, including autoimmune salivary infiltration, ard unaffected. Furthermore, contrary to previous studies, cyclophosphamide administration was unable to induce diabetes in treated mice which suggests that deletion or anergy might be the mechanism by which neonatal intrathymic islet injection protects from disease. However, anti-islet antigen antibodies were still present in these mice which suggests that the mechanism of disease protection may be more complex.

    View details for Web of Science ID A1994PL69600001

    View details for PubMedID 7840850

  • CD4-POSITIVE HEAT-STABLE ANTIGEN-POSITIVE THYMOCYTES CAUSE GRAFT-VERSUS-HOST DISEASE ACROSS NONMAJOR HISTOCOMPATIBILITY COMPLEX INCOMPATIBILITIES EUROPEAN JOURNAL OF IMMUNOLOGY Charlton, B., Meltzer, J., Fathman, C. G. 1994; 24 (7): 1706-1709

    Abstract

    Single-positive thymocytes are the immediate precursors of peripheral recent thymic emigrants (RTE) which develop into mature peripheral T cells. The functional ability of RTE is unclear but their state of differentiation may be relevant to the development of tolerance to peripheral "self" antigens. Since RTE are difficult to analyze, precursor CD4+/8- thymocytes were assessed in a model in vivo to determine their functional capability and their susceptibility to tolerance induction. The ability of both heat-stable antigen-positive (HSA+) (immature) and HSA- (mature) single-positive thymocytes to cause graft-versus-host disease (GVHD) across non-major histocompatibility complex differences were examined. Both HSA- and HSA+ CD4+/8- thymocytes from C3H mice caused lethal GVHD in AKR recipients as did CD4+ peripheral T cells in controls. Further, neonatal C3H thymocytes also caused lethal GVHD in AKR recipients. Since CD4+/8- thymocytes are the precursors of RTE, these results suggest that RTE are not susceptible to tolerance induced to "minor" antigens and may have a normal immune function in vivo. This would suggest that peripheral tolerance may be dependent upon the manner of antigen presentation rather than T cell maturity.

    View details for Web of Science ID A1994NX26100036

    View details for PubMedID 7913040

  • IMMUNOTHERAPEUTIC STRATEGIES DIRECTED AT THE TRIMOLECULAR COMPLEX ADVANCES IN IMMUNOLOGY, VOLUME 56 Gaur, A., Fathman, C. G. 1994; 56: 219-265

    View details for Web of Science ID A1994BB09D00005

    View details for PubMedID 7521115

  • MECHANISMS OF TRANSPLANTATION TOLERANCE ANNUAL REVIEW OF IMMUNOLOGY Charlton, B., Auchincloss, H., Fathman, C. G. 1994; 12: 707-734

    Abstract

    Transplantation tolerance, the long-term acceptance of grafted tissue in the absence of continuous immunosuppression, remains an elusive goal in humans, but it has been achieved in animal models using numerous approaches. The mechanisms behind graft acceptance vary according to the means used to create the state of acceptance. Several major mechanisms can now be recognized. While thymic deletion of T cells appears to be a mainstay of self-tolerance, its role in transplantation tolerance now seems to be less significant. In contrast, extrathymic mechanisms of transplantation tolerance seem to be major factors in long-term graft acceptance. If donor antigens are presented in a nonimmunogenic manner on the graft, e.g. due to modification of graft tissue by culture, peripheral T cells of the recipient may ignore the graft. Alternatively, nonstimulatory presentation of donor antigens on graft tissue can induce a state of unresponsiveness in recipient T cells, i.e. anergy, rather than activating them to destroy the graft. Suppression mechanisms also operate to control graft rejection and may be specific or nonspecific in nature. Specific suppression mechanisms might act in an idiotype or antigen-specific fashion, and evidence is accumulating that this may be mediated through the elaboration of cytokines. Donor antigen-specific T cells may be activated to produce "protective" cytokines which then regulate the generation of destructive T cells. Future therapies will be aimed at affecting graft acceptance through these peripheral mechanisms.

    View details for Web of Science ID A1994NF48700022

    View details for PubMedID 8011295

  • SELECTION FOR AMINO-ACID-SEQUENCE AND J-BETA-ELEMENT USAGE IN THE BETA-CHAIN OF DBA 2V BETA(B)-DERIVED AND DBA 2V BETA(A)-DERIVED MYOGLOBIN-SPECIFIC T-CELL CLONES JOURNAL OF IMMUNOLOGY Ruberti, G., Paragas, V., Kim, D., Fathman, C. G. 1993; 151 (11): 6185-6194

    Abstract

    Previous experiments in our laboratory have demonstrated that there is a marked restriction on the TCR beta-chain usage in DBA/2 mice in response to the sperm whale myoglobin (SpWMb) determinant 110-121, predominated by the use of the V beta 8.2 gene element. We analyzed the response of mice that had been genetically depleted of V beta 8+ T cells by generating a DBA/2 line that carries the V beta a TCR haplotype. Despite the very limited TCR repertoire expressed by DBA/2V beta a mice, they made an excellent response after immunization with the SpWMb 110-121 peptide. Data presented in this manuscript demonstrate that there is an equally restricted TCR V beta-chain utilization in the T-cell response to the determinant SpWMb 110-121 in DBA/2V beta a mice. Unexpectedly, there was a shift of MHC restriction of this determinant to T cells in the V beta a strain when compared with the V beta b strain of DBA/2 mice. We had previously demonstrated that DBA/2 mice utilized both the hybrid E alpha dA beta d MHC molecule as well as the conventional A alpha dA beta d molecule as presenting elements in response to SpWMb 110-121. Data presented in this manuscript demonstrate that the T-cell response in DBA/2V beta a mice is entirely restricted by the A alpha dA beta d MHC class II molecule. By analyzing a panel of SpWMb 110-121-specific T-cell clones from DBA/2V beta a mice, we were able to study the TCR repertoire expressed on T cells from mice that lack the V beta 8.2 gene. The V beta usage by the panel of clones analyzed was remarkably homogeneous. Thirteen of the 17 clones analyzed used the V beta 1 gene segment. Perhaps more striking was the junctional region nucleotide and amino acid sequences that were shared among these clones and that were similar to the V beta 8.2 clones analyzed previously. All clones assayed used the J beta 2.6 element, as did the great majority of the V beta 8.2 clones analyzed from DBA/2 (and B10.D2) V beta b mice. Importantly, in each strain of mice, irrespective of the V beta utilized, each TCR appeared to have selected an acidic amino acid in the beta-chain at position 100.(ABSTRACT TRUNCATED AT 400 WORDS)

    View details for Web of Science ID A1993MH75600031

    View details for PubMedID 8245460

  • INDUCTION OF RELAPSING PARALYSIS IN EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS BY BACTERIAL SUPERANTIGEN NATURE Brocke, S., Gaur, A., Piercy, C., Gautam, A., Gijbels, K., Fathman, C. G., Steinman, L. 1993; 365 (6447): 642-644

    Abstract

    The role of infection in the pathogenesis of clinical relapses that occur in most autoimmune diseases, including multiple sclerosis, remains to be established. Experimental autoimmune encephalomyelitis (EAE) serves as a model for multiple sclerosis, with episodes of relapsing paralysis. In certain strains of mice, T-lymphocytes expressing the V beta 8 T-cell receptor (TCR) engage the amino-terminal epitope Ac1-11 of myelin basic protein, leading to EAE. The bacterial superantigen staphylococcal enterotoxin B (SEB) activates V beta 8-expressing T cells. Here we show that after immunization with Ac1-11, or after transfer of encephalitogenic T-cell lines or clones reactive to Ac1-11, SEB induces exacerbation or relapses of paralytic disease in mice that are in clinical remission following an initial episode of paralysis, and triggers paralysis in mice with subclinical disease. Tumour necrosis factor has a critical role in the mechanism underlying SEB-induced exacerbation of disease, because anti-tumour necrosis factor antibody given in vivo delays the onset of paralysis triggered by SEB. On reactivation of autoaggressive cells through their T-cell receptor, superantigens may induce clinical relapses of autoimmune disease.

    View details for Web of Science ID A1993MB84600057

    View details for PubMedID 7692305

  • EVIDENCE THAT CLONAL ANERGY IS INDUCED IN THYMIC MIGRANT CELLS AFTER ANTI-CD4-MEDIATED TRANSPLANTATION TOLERANCE TRANSPLANTATION Alters, S. E., Song, H. K., Fathman, C. G. 1993; 56 (3): 633-638

    Abstract

    Diabetic (B6) (IE-) mice treated with a depleting regimen of anti-CD4 monoclonal antibody at the time of transplantation with A/J (IEK) islets of Langerhans showed indefinite acceptance of their islet allograft, as evidenced by persistent normoglycemia. To address the mechanisms involved in such anti-CD4 induced transplantation tolerance we studied potentially IE-reactive V beta 11+ T cells from the tolerant allografted mice. Following complete repopulation of the CD4+ cells, both the CD4+V beta 11+ and CD8+V beta 11+ T cell subsets of the transplanted mice were unresponsive to anti-V beta 11 specific crosslinking. In contrast, lymphocytes tested within the first ten days following transplant were responsive to anti-V beta 11 specific crosslinking; this response decreased as a function of time and reached background levels by day 120 posttransplant. Sorting experiments indicated that the response of lymphocytes to anti-V beta 11 specific crosslinking seen during the initial 120 days posttransplant was confined to the peripheral CD8+ cells; the repopulating CD4+V beta 11+ T cells were unresponsive. In addition, administration of r-IL-2 at the time of transplantation induced rejection in anti-CD4-treated animals, again indicating that the peripheral CD8+ cells could respond shortly after transplant if provided with appropriate help. The decreasing response of CD8+ T cells from transplanted animals to anti-V beta 11 stimulation was inversely correlated with the rate of migration of cells from the thymus to the periphery, implying that new thymic migrant V beta 11+ cells, both CD4+ and CD8+, were rendered anergic upon encountering peripheral alloantigen. These data suggest the possibility that recent thymic migrants are rendered anergic upon encountering antigen in the periphery, a simple model to serve as a "fail-safe" mechanism to prevent autoreactivity.

    View details for Web of Science ID A1993LZ87000027

    View details for PubMedID 8212161

  • SEB INDUCED ANERGY - MODULATION OF IMMUNE-RESPONSE TO T-CELL DETERMINANTS OF MYOGLOBIN AND MYELIN BASIC-PROTEIN JOURNAL OF IMMUNOLOGY Gaur, A., Fathman, C. G., Steinman, L., Brocke, S. 1993; 150 (7): 3062-3069

    Abstract

    Superantigens have the ability to stimulate a subset of T cells based upon their expressed TCR beta-chain. It has been demonstrated that the administration of staphylococcal enterotoxin B (SEB) in mice leads to unresponsiveness in V beta 8+ T cells in vivo which are the same T cells that could be stimulated in vitro by this enterotoxin. We present here data on the effect of SEB administration in DBA/2 and (PL/J x SJL)F1 mice on their T cell response to two different T cell determinants, the responses against which are dominated by the use of V beta 8+ T cells. Treatment of mice with SEB not only diminished their primary T cell proliferative response to these determinants, but also was able to effectively reduce the memory T cell response. SEB treatment, however, showed only a modest effect in preventing Ac 1-11-induced experimental autoimmune encephalomyelitis in H-2u mice.

    View details for Web of Science ID A1993KT80500051

    View details for PubMedID 7681087

  • THE USE OF GRANZYME A AS A MARKER OF HEART-TRANSPLANT REJECTION IN CYCLOSPORINE OR ANTI-CD4 MONOCLONAL ANTIBODY-TREATED RATS TRANSPLANTATION CHEN, R. H., IVENS, K. W., Alpert, S., Billingham, M. E., Fathman, C. G., Flavin, T. F., Shizuru, J. A., Starnes, V. A., Weissman, I. L., Griffiths, G. M. 1993; 55 (1): 146-153

    Abstract

    Granzyme A is a serine protease expressed by populations of human and mouse natural killer cells and activated CD4+ and CD8+ cytotoxic lymphocytes; its expression marks a subset of inflammatory cells in allografts, autoimmune diabetes, and a number of other inflammatory lesions. In order to describe more completely the correlation between granzyme A expression and the presence of in vivo cytolytic effects, we grafted allogeneic rat hearts with vascular anastomoses in a heterotopic location, and treated the hosts with either cyclosporine, anti-CD4 monoclonal antibody (MRC OX38), or no therapy. The grafts were evaluated by palpation for cardiac functions, by immunohistochemistry for CD4/CD8 expression, by hematoxylin-and-eosin staining for inflammatory infiltration, and by in situ hybridization for granzyme A expression. The appearance of granzyme A+ cells in untreated allografts preceded both functional and standard histopathological and immunohistochemical evidence of graft rejection by two days. In donor-recipient combinations where cyclosporine and anti-CD4 treatments allowed indefinite allograft survival, the allografts showed minimal numbers of granzyme A+ cells, whether cellular infiltrates developed or not. The number of granzyme A+ cells present in the cardiac allografts in treated and untreated animals correlated with either current or impending episodes of rejection. The early time course of granzyme A expression suggests that it can be used as an early and reliable marker of graft rejection.

    View details for Web of Science ID A1993KH66000027

    View details for PubMedID 8420039

  • REQUIREMENT FOR CD8+ CELLS IN T-CELL RECEPTOR PEPTIDE-INDUCED CLONAL UNRESPONSIVENESS SCIENCE Gaur, A., Haspel, R., Mayer, J. P., Fathman, C. G. 1993; 259 (5091): 91-94

    Abstract

    T cell receptor (TCR) vaccination in rats prevents the development of experimental allergic encephalomyelitis (EAE), an animal model of multiple sclerosis. The mechanism of this potential immunotherapy was examined by vaccinating mice with an immunogenic peptide fragment of the variable region of the TCR V beta 8.2 gene. Another immunogen that usually induces an immune response mediated by V beta 8.2+ T cells was subsequently inhibited because specific clonal unresponsiveness (anergy) had been induced. Depletion of CD8+ cells before TCR peptide vaccination blocked such inhibition. Thus, the clonal anergy was dependent on CD8+ T cells, and such immunoregulatory T cells may participate in the normal course of EAE.

    View details for Web of Science ID A1993KE60100036

    View details for PubMedID 8418501

  • EVIDENCE THAT ANTI-CD8 ABROGATES ANTI-CD4 MEDIATED CLONAL ANERGY BUT ALLOWS ALLOGRAFT SURVIVAL IN MICE TRANSPLANTATION Song, H. K., Alters, S. E., Fathman, C. G. 1993; 55 (1): 133-139

    Abstract

    Monoclonal antibodies directed against different T cell subpopulations have been used in several rodent models of transplantation to induce long-term unresponsiveness to allografts by a variety of mechanisms. To investigate whether different mechanisms may be operative when different regimens of mAb therapy are used, we studied the effects of various combinations of anti-T-cell antibody treatment on the induction of tolerance in a mouse islet allograft model. Anti-CD4 mAb alone, anti-CD8 mAb alone, anti-CD4 mAb plus anti-CD8 mAb, and anti-Thy1.2 mAb alone were given at the time of engraftment. Only the anti-CD4 mAb and the anti-CD4 mAb plus anti-CD8 mAb regimens were successful in inducing permanent unresponsiveness to islet allografts. We have previously shown that anti-CD4 mAb alone induces permanent unresponsiveness to islet allografts by a mechanism of clonal anergy, as demonstrated by unresponsiveness of potentially alloreactive T cells to anti-T-cell receptor-specific cross-linking. Interestingly, the potentially alloreactive T cell subsets of recipient mice (V beta 5+ and V beta 11+) made unresponsive to islet allografts by anti-CD4 mAb plus anti-CD8 mAb therapy were not found to be anergic using the same assay. Differences between the repopulation kinetics of CD8+ T cells of anti-CD4 mAb plus anti-CD8 mAb treated recipient mice, which accepted islet allografts, and anti-Thy1.2 treated recipient mice, which rejected islet allografts despite similar levels of initial T cell depletion, suggest that unresponsiveness to alloantigen may have been induced in anti-CD4 mAb plus anti-CD8 mAb treated recipients by clearance of donor passenger leukocytes during prolonged CD8+ T cell depletion.

    View details for Web of Science ID A1993KH66000025

    View details for PubMedID 8420037

  • Anti-CD4 antibodies in diabetes. Immunology series Shizuru, J. A., Fathman, C. G. 1993; 59: 237-252

    View details for PubMedID 8096401

  • AMELIORATION OF AUTOIMMUNE ENCEPHALOMYELITIS BY MYELIN BASIC-PROTEIN SYNTHETIC PEPTIDE INDUCED ANERGY SCIENCE Gaur, A., WIERS, B., Liu, A., Rothbard, J., Fathman, C. G. 1992; 258 (5087): 1491-1494

    Abstract

    Experimental autoimmune encephalomyelitis (EAE), a demyelinating disease of the central nervous system that can be induced in susceptible strains of mice by immunization with myelin basic protein (MBP) or its immunodominant T cell determinants, serves as a model of human multiple sclerosis. Tolerance to MBP in adult mice was induced by intraperitoneal injection of synthetic peptides of immunodominant determinants of MBP and prevented MBP-induced EAE. Furthermore, tolerance-inducing regimens of peptides administered to mice after the disease had begun (10 days after induction with MBP) blocked the progression and decreased the severity of EAE. Peptide-induced tolerance resulted from the induction of anergy in proliferative, antigen-specific T cells.

    View details for Web of Science ID A1992JZ62500031

    View details for PubMedID 1279812

  • IMMUNOTHERAPY OF RHEUMATIC DISEASES BASED ON UNDERSTANDING GENETIC PREDISPOSITION TO THE DEVELOPMENT OF THESE DISEASES RHEUMATIC DISEASE CLINICS OF NORTH AMERICA Fathman, C. G. 1992; 18 (4): 915-926

    Abstract

    During the past two decades, investigators have made great inroads in understanding markers of genetic predisposition to the development of rheumatic diseases. An important question that must be addressed by investigators and clinicians is whether this knowledge will ultimately benefit patients, either through genetic counseling or predictions for beneficial therapeutic intervention. This article discusses various disease mechanisms and modes of immunotherapy such as anti-CD4, major histocompatibility complex blockade, and T-cell receptor-based and determinant-induced responsiveness.

    View details for Web of Science ID A1992LA28000012

    View details for PubMedID 1280848

  • ANTI-CD4 MONOCLONAL-ANTIBODIES IN THERAPY - CREATION OF NONCLASSICAL TOLERANCE IN THE ADULT IMMUNOLOGICAL REVIEWS Shizuru, J. A., Alters, S. E., Fathman, C. G. 1992; 129: 105-130

    View details for Web of Science ID A1992JZ99700005

    View details for PubMedID 1361179

  • LIMITED T-CELL RECEPTOR BETA-CHAIN USAGE IN THE SPERM WHALE MYOGLOBIN 110-121/E-ALPHA(D)A-BETA(D) RESPONSE BY H-2(D) CONGENIC MOUSE STRAINS JOURNAL OF IMMUNOLOGY Sellins, K. S., Danska, J. S., Paragas, V., Fathman, C. G. 1992; 149 (7): 2323-2327

    Abstract

    The specificity and TCR gene usage of a panel of sperm whale myoglobin (SpWMb)-reactive T cell clones from DBA/2 mice have previously been characterized, to study structure-function relationships between components of the ternary complex consisting of Ag, TCR, and MHC class II molecules, whose interaction leads to Th cell activation. These DBA/2 clones were specific for epitopes within the residue 110 to 121 region of SpWMb, in the context of the mixed isotype molecule E alpha dA beta d, and expressed the TCR V beta 8.2 gene element. SpWMb-specific T cell hybridomas from the H-2d-congenic B10.D2 mouse strain, which differs from the DBA/2 strain only in the non-MHC background, were generated and compared with the T cell hybridomas from DBA/2 mice, in order to investigate the influence of non-MHC genes on the specificity of the T cell response to the 110-121 epitope. V beta usage by these hybridomas was very homogeneous; three of three DBA/2 and eight of nine B10.D2 hybridomas specific for the 110-121 epitope, in the context of the mixed isotype molecule E alpha dA beta d, expressed the V beta 8.2 gene product. Nucleotide and amino acid sequences of D beta, J beta, and N regions were also similar. One 110-121/E alpha dA beta d-specific B10.D2 hybridoma used V beta 7, a V beta that is clonally deleted in DBA/2 mice. These experiments suggest that a similar set of TCR beta genes are used to respond to a given epitope, regardless of non-MHC background, and they support the hypothesis that, despite great variability between individuals in their non-MHC background genes, human HLA-associated diseases might result from the formation of a particular ternary complex consisting of a shared MHC molecule, a common "disease-associated" epitope, and a shared TCR.

    View details for Web of Science ID A1992JP18900012

    View details for PubMedID 1382096

  • What a rheumatologist needs to know about T cell receptor structure and function. journal of rheumatology. Supplement Fathman, C. G. 1992; 32: 12-15

    Abstract

    By understanding normal immune response, it has been possible to develop therapeutic strategies toward the treatment of autoimmune disease. The association of autoimmune disease with the major histocompatibility complex (MHC) class II gene products suggests that the inductive events in which the putative autoantigen is presented on the surface of antigen presenting cells in the context of the MHC class II gene products and is recognized by CD4(+) helper or inducer T cells form an interesting target for immunotherapeutic intervention. By understanding the structure/function relationships of T cell receptors for antigen, it might be possible to develop novel immunotherapeutic strategies for the treatment of seropositive rheumatoid arthritis. Studies described below review recent progress in understanding the components of the ternary complex and suggest possible areas of immunotherapeutic intervention.

    View details for PubMedID 1613729

  • PRESENTATION OF ANTIGEN BY MIXED ISOTYPE CLASS-II MOLECULES IN NORMAL H-2D MICE JOURNAL OF EXPERIMENTAL MEDICINE Ruberti, G., Sellins, K. S., Hill, C. M., Germain, R. N., Fathman, C. G., Livingstone, A. 1992; 175 (1): 157-162

    Abstract

    A panel of DBA/2 T cell hybridomas specific for the sperm whale myoglobin epitope 110-121 was found to recognize antigen presented by the mixed isotype class II molecule E alpha dA beta d. The response was blocked by monoclonal antibodies specific for E alpha and A beta d chains; in addition, the hybridomas responded to antigen presented by L cells expressing E alpha A beta d molecules, and made no response with L cells expressing I-Ad or I-Ed molecules. Two more groups of hybridomas isolated from DBA/2 and B10.D2 mice immunized with myoglobin also recognized peptide 110-121 presented by E alpha d A beta d. Thus, although it is expressed at biochemically undetectable levels on spleen cells, the E alpha d A beta d molecule is an important presenting element in normal H-2d mice making a conventional immune response to a protein antigen. These results suggest that high levels of class II expression are not a prerequisite for T cell activation.

    View details for Web of Science ID A1992GY43600019

    View details for PubMedID 1730914

  • ANTI-CD8 ABROGATES EFFECT OF ANTI-CD4-MEDIATED ISLET ALLOGRAFT SURVIVAL IN RAT MODEL DIABETES SEYDEL, K., Shizuru, J., Grossman, D., Wu, A., Alters, S., Fathman, C. G. 1991; 40 (11): 1430-1434

    Abstract

    We studied the effects of anti-CD4 treatment of diabetic ACI rats on the induction of tolerance to allogeneic (Lewis) islet allografts. When given as a 4-day treatment regimen, OX38, a mouse anti-rat CD4 antibody, caused depletion of greater than 80% of CD4+ cells from the peripheral blood of treated rats. After induction of diabetes (a single high-dose bolus of streptozocin) and 3 days after the initiation of anti-CD4 immunotherapy, recipient ACI rats were transplanted with fully allogeneic (Lewis) islets of Langerhans via the portal circulation. These transplanted islets were capable of returning the anti-CD4-treated ACI recipients to normoglycemia, which was maintained indefinitely in the absence of further immunosuppression. In contrast, treatment of recipient rats with OX8, an anti-CD8 monoclonal antibody (MoAb), induced only a slight prolongation of graft survival (less than or equal to 30 days). Further characterization of the cellular requirements for the induction of long-term transplantation survival revealed that successful pretransplantation anti-CD4 therapy could be ablated by the coincident treatment of recipient rats with depleting levels of anti-CD8 MoAb. These data point to the necessity of a regulator CD8+ cell in the induction of anti-CD4-mediated transplantation survival in this rat model of islet transplantation.

    View details for Web of Science ID A1991GN93200010

    View details for PubMedID 1834499

  • GENETIC DISSECTION OF T-CELL RECEPTOR-V-BETA GENE REQUIREMENTS FOR SPONTANEOUS MURINE DIABETES JOURNAL OF EXPERIMENTAL MEDICINE Shizuru, J. A., TAYLOREDWARDS, C., Livingstone, A., Fathman, C. G. 1991; 174 (3): 633-638

    Abstract

    It has been demonstrated, in certain autoimmune disease models, that pathogenic T cells express antigen receptors of limited diversity. It has been suggested that the T cells responsible for the pathogenesis of type I diabetes mellitus might similarly demonstrate restricted T cell receptor (TCR) usage. Recently, attempts have been made to identify the V beta subset(s) that initiates and/or perpetuates the antiislet response in a mouse model of spontaneous autoimmune diabetes (non-obese diabetic [NOD] mice). In studies reported here, we have bred NOD mice to a mouse strain that congenitally lacks approximately one-half of the conventional TCR V beta alleles. Included in this deletion are TCR V beta gene products previously implicated as being involved in the pathogenesis of NOD disease. By studying second backcross-intercross animals, we were able to demonstrate that this deletion of TCR V beta gene segments did not prevent the development of insulitis or diabetes.

    View details for Web of Science ID A1991GC96000014

    View details for PubMedID 1831491

  • THE T-CELL RECEPTOR REPERTOIRE INFLUENCES V-BETA ELEMENT USAGE IN RESPONSE TO MYOGLOBIN JOURNAL OF EXPERIMENTAL MEDICINE Ruberti, G., Gaur, A., Fathman, C. G., Livingstone, A. M. 1991; 174 (1): 83-92

    Abstract

    T cell clones recognizing the sperm whale myoglobin (SpWMb) epitope 110-121 in association with H-2d major histocompatibility complex class II molecules display a very limited heterogeneity of T cell receptor (TCR) V beta usage in DBA/2 mice. All clones previously tested used the same V beta 8.2 gene segment and very restricted junctional regions. To investigate the significance of this observation in vivo, we immunized DBA/2 mice with the intact SpW Mb protein or peptide 110-121. Only the V beta 8+ T cells showed any significant response to the 110-121 epitope. The response to peptide 110-121 was then analyzed in mice which, either as a consequence of antibody depletion or through genetic deletion of TCR V beta genes, lacked V beta 8+ peripheral T cells. DBA/2 mice depleted of V beta 8+ T cells by antibody treatment responded poorly to the 110-121 peptide, and only at high antigen concentrations. In contrast, DBA/2V beta a mice (homozygous for a deletion of multiple V beta gene segments including the V beta 8 family) made a response at least as great as that made by DBA/2 mice, even though the DBA/2V beta a mice had a very restricted TCR V beta repertoire compared with DBA/2 mice. Mechanisms which might determine differences in the 110-121 specific response of DBA/2, DBA/2V beta a and F23.1-treated DBA/2 mice are discussed.

    View details for Web of Science ID A1991FU89700011

    View details for PubMedID 2056283

  • MOLECULAR ANALYSIS OF THE ROLE OF THE HLA CLASS-II GENES DRB1, DQA1, DQB1, AND DPB1 IN SUSCEPTIBILITY TO LYME ARTHRITIS HUMAN IMMUNOLOGY Ruberti, G., Begovich, A. B., STEERE, A. C., Klitz, W., Erlich, H. A., Fathman, C. G. 1991; 31 (1): 20-27

    Abstract

    Using the polymerase chain reaction and sequence-specific oligonucleotide probes, we have determined the distribution of the alleles at the polymorphic class II loci, DRB1, DQA1, DQB1, and DPB1 in 25 patients with Lyme arthritis. Comparison of the frequencies of the DRB1 and DPB1 alleles in Lyme arthritis patients to the Centre d'Etude du Polymorphisme Humaine control panel revealed statistically significant differences between the two groups. An increase in DRB1*1301 is noted along with a unique distribution of the DR4 subtypes within the 9 DR4+ patients. In addition, an increase in the rate DPB1*1001 and the more common DPB1*0201 alleles is reported.

    View details for Web of Science ID A1991FH13300004

    View details for PubMedID 1679052

  • ANTI-CD4 MEDIATES CLONAL ANERGY DURING TRANSPLANTATION TOLERANCE INDUCTION JOURNAL OF EXPERIMENTAL MEDICINE Alters, S. E., Shizuru, J. A., Ackerman, J., Grossman, D., Seydel, K. B., Fathman, C. G. 1991; 173 (2): 491-494

    Abstract

    Depletion of CD4+ cells using anti-CD4 monoclonal antibodies leads to allograft tolerance. Here we show that anti-CD4-mediated tolerance to pancreatic islets of Langerhans transplanted from an A/J (IEk) donor to a diabetic C57B1/6 (B6) (IE-) recipient occurs in the absence of clonal deletion of the potentially IE-reactive V beta 11+ T cells. Instead, a state of clonal anergy is induced in both the CD4+V beta 11+ and CD8+V beta 11+ T cell subsets. This clonal anergy can be partially overcome in vitro by the addition of recombinant interleukin 2.

    View details for Web of Science ID A1991EV17500025

    View details for PubMedID 1899105

  • GENETIC-ANALYSIS OF DIABETES IN THE NONOBESE DIABETIC MOUSE .1. MHC AND T-CELL RECEPTOR BETA-GENE EXPRESSION JOURNAL OF IMMUNOLOGY Livingstone, A., Edwards, C. T., Shizuru, J. A., Fathman, C. G. 1991; 146 (2): 529-534

    Abstract

    Backcross nonobese diabetic (NOD) ((NOD x SWr)F1 x NOD) mice (108 females and 105 males) were typed for MHC, TCR V beta, and monitored for 350 days for the onset of diabetes. The presence of "antipolar" antibodies in the sera and the occurrence of insulitis was examined in a proportion of these backcross mice. There was no difference in the incidence of diabetes in mice heterozygous for TCR V beta b/a vs those homozygous for TCR V beta b/b. Among the 17 diabetics (all female) detected in this backcross, 14/17 were H-2nod/nod but 3/17 were H-2nod/q. This supports a previous observation suggesting that the MHC-linked diabetogenic gene originally thought to be recessive may rather be dominant but have a low penetrance in the heterozygous state. Antipolar autoantibodies were found in both female and male backcross mice, and were similarly distributed in diabetic and nondiabetic mice. There appeared to be no correlation between the level of these auto-antibodies and development of diabetes. The incidence and severity of insulitis was linked to MHC but no influence of TCR genes on insulitis nor an association between insulitis and antipolar antibodies could be demonstrated in this study. Further analyses of H-2nod/nod intercross mice homozygous for TCR V beta a or TCR V beta b are currently underway.

    View details for Web of Science ID A1991EU50500019

    View details for PubMedID 1824775

  • DRB1-STAR-LY10 - A NEW DRB1 ALLELE AND ITS HAPLOTYPIC ASSOCIATION IMMUNOGENETICS MCCLURE, G. R., Ruberti, G., Fathman, C. G., Erlich, H. A., Begovich, A. B. 1990; 32 (3): 214-217

    View details for Web of Science ID A1990ED27600011

    View details for PubMedID 2228048

  • T-CELL RECEPTOR-MEDIATED NEGATIVE SELECTION OF AUTOREACTIVE LYMPHOCYTE-T PRECURSORS OCCURS AFTER COMMITMENT TO THE CD4 OR CD8 LINEAGES JOURNAL OF EXPERIMENTAL MEDICINE Guidos, C. J., Danska, J. S., Fathman, C. G., Weissman, I. L. 1990; 172 (3): 835-845

    Abstract

    To identify the maturational stage(s) during which T cell receptor (TCR)-mediated positive and negative selection occurs, we followed the development of CD4+8- and CD4-8+ T cells from TCRlo CD4+8+ thymic blasts in the presence of different positive and negative selecting (major histocompatibility complex or Mls) elements. We describe novel lineage-committed transitional intermediates that are TCRmed CD4+8lo or TCRmed CD4lo8+, and that show evidence of having been positively selected. Furthermore, negative selection is not evident until after cells have attained one of the TCRmed transitional phenotypes. Accordingly, we propose that negative selection in normal mice occurs only after TCRlo CD4+8+ precursors have been positively selected into either the CD4 or CD8 lineage.

    View details for Web of Science ID A1990DW49100018

    View details for PubMedID 2143774

  • INDUCTION OF DONOR-SPECIFIC UNRESPONSIVENESS TO CARDIAC ALLOGRAFTS IN RATS BY PRETRANSPLANT ANTI-CD4 MONOCLONAL-ANTIBODY THERAPY TRANSPLANTATION Shizuru, J. A., Seydel, K. B., Flavin, T. F., Wu, A. P., Kong, C. C., Hoyt, E. G., Fujimoto, N., Billingham, M. E., Starnes, V. A., Fathman, C. G. 1990; 50 (3): 366-373

    Abstract

    In the present report a monoclonal antibody designated OX-38 directed against the rat CD4 molecule was tested for its ability to prolong the survival of heterotopic vascularized rat heart allografts transplanted across major histocompatibility barriers. Fluorescence-activated cell-sorter analysis showed that administration of OX-38 selectively depleted 80-95% of CD4+ cells from peripheral blood of treated rats. The immunosuppressive effects of OX-38 in vivo were verified by suppression of an antibody response against OX-38 itself as a heterologous protein immunogen. Recipient rats received OX-38 antibody as a single agent given in pretransplant regimens. Nine of 12 treated rats have maintained heterotopic abdominal heart allografts for greater than 175 days. Control rats that did not receive antibody therapy rejected their grafts within 14 days. Rats that maintained heart allografts for greater than 100 days accepted second donor strain hearts but rejected third-party heart grafts transplanted into the femoral space. Anti-CD4-induced allograft unresponsiveness persisted for at least 90 days following surgical removal of donor tissue and retransplantation of a second donor-matched heart. These results indicated that transient, pretransplant therapy with monoclonal antibodies directed against the CD4+ lymphocyte induced specific, long-lasting unresponsiveness to fully MHC-mismatched cardiac allografts in rats without additional immunosuppression.

    View details for Web of Science ID A1990DY85500002

    View details for PubMedID 1976282

  • THE PRESUMPTIVE CDR3 REGIONS OF BOTH T-CELL RECEPTOR ALPHA-CHAIN AND BETA-CHAIN DETERMINE T-CELL SPECIFICITY FOR MYOGLOBIN PEPTIDES JOURNAL OF EXPERIMENTAL MEDICINE Danska, J. S., Livingstone, A. M., Paragas, V., Ishihara, T., Fathman, C. G. 1990; 172 (1): 27-33

    Abstract

    The T cell receptor alpha/beta (TCR-alpha/beta) is encoded by variable (V), diversity (D), joining (J), and constant (C) segments assembled by recombination during thymocyte maturation to produce a heterodimer that imparts antigenic specificity to the T cell. Unlike immunoglobulins (Igs), which bind free antigen, the ligands of TCR-alpha/beta are cell surface complexes of intracellularly degraded antigens (i.e., peptides) bound to and presented by polymorphic products of the major histocompatibility complex (MHC). Therefore, antigen recognition by T cells is defined as MHC restricted. A model has been formulated based upon the similarity between TCR-alpha/beta V region and Ig Fab amino acid sequences, and the crystal structure of the MHC class I and Ig molecules. This model predicts that the complementarity determining regions (CDR) 1 and 2, composed of TCR V alpha and V beta segments, primarily contact residues of the MHC alpha helices, whereas V/J alpha and V/D/J beta junctional regions (the CDR3 equivalent) contact the peptide in the MHC binding groove. Because polymorphism in MHC proteins is limited relative to the enormous diversity of antigenic peptides, the TCR may have evolved to position the highly diverse junctional residues (CDR3), where they have maximal contact with antigen bound in the MHC peptide groove. Here, we demonstrate a definitive association between CDR3 sequences in both TCR alpha and beta chains, and differences in recognition of antigen fine specificity using a panel of I-Ed-restricted, myoglobin-reactive T cell clones. Acquisition of these data relied in part upon a modification of the polymerase chain reaction that uses a degenerate, consensus primer to amplify TCR alpha chains without foreknowledge of the V alpha segments they utilize.

    View details for Web of Science ID A1990DL09100005

    View details for PubMedID 1694219

  • SHARED MOLECULAR MARKERS OF GENETIC PREDISPOSITION TO SEROPOSITIVE RHEUMATOID-ARTHRITIS HUMAN IMMUNOLOGY Morel, P. A., HORN, G. T., Budd, R. C., Erlich, H. A., Fathman, C. G. 1990; 27 (2): 90-99

    Abstract

    Rheumatoid arthritis is associated with the human class II major histocompatibility complex antigens known as HLA-DR4. HLA-DR4 can be subdivided by cellular typing into five subtypes: Dw4, Dw10, Dw13, Dw14, and Dw15. By traditional serologic methods, 60-80% of rheumatoid arthritis patients type HLA-DR4 compared to approximately 20% of the general population. It has been demonstrated, using a panel of four alloreactive T-cell clones, each of which recognized HLA-DR4, Dw14 homozygous typing cells, that cells from all of a group of 23 rheumatoid arthritis patients could be recognized by one or more of these clones regardless of the patients' serologic typing. As the expressed polymorphism of the DR molecule is accounted for by the beta 1 gene, this gene was amplified, using the polymerase chain reaction, and sequenced. Seven patients whose cells were recognized by one of the DR4, DW14-specific T-cell clones, T431, were analyzed. All of these patients shared a common sequence in the third hypervariable region of the DR beta 1 chain gene. The sequence identified is the one normally associated with DR4, Dw14 and DR1. Patients and DR4-positive controls whose cells did not stimulate this clone did not share this sequence. These results suggest that this hypervariable region might be an important contribution to a restriction site for the putative causative agent(s) in rheumatoid arthritis.

    View details for Web of Science ID A1990CJ52400003

    View details for PubMedID 2298612

  • CARDIAC ALLOGRAFT PROLONGATION IN MICE TREATED WITH COMBINED POSTTRANSPLANTATION TOTAL-LYMPHOID IRRADIATION AND ANTI-L3T4 ANTIBODY THERAPY TRANSPLANTATION TRAGER, D. K., Banks, B. A., Rosenbaum, G. E., HOLM, B. I., Shizuru, J. A., Strober, S., Fathman, C. G. 1989; 47 (4): 587-591

    Abstract

    Neonatal cardiac allograft survival was examined in mice treated with anti-L3T4 antibody, posttransplantation total lymphoid irradiation (TLI) or a combination of both therapies. Independently, both posttransplantation TLI and short-course antibody treatment allowed minimal prolongation. However, synergistic prolongation in graft survival was observed with the combination (synergistic) therapy. Fluorescence-activated cell sorter analysis of peripheral blood lymphocytes from animals treated with combined anti-L3T4 and posttransplantation TLI additionally revealed "synergy" with respect to the degree of peripheral lymphocyte depletion.

    View details for Web of Science ID A1989U208300004

    View details for PubMedID 2523098

  • A SUBSET OF MEMORY CD4+ HELPER LYMPHOCYTES-T IDENTIFIED BY EXPRESSION OF PGP-1 JOURNAL OF EXPERIMENTAL MEDICINE Butterfield, K., Fathman, C. G., Budd, R. C. 1989; 169 (4): 1461-1466

    Abstract

    The Pgp-1 glycoprotein (Ly-24 antigen) is acquired by mature murine T lymphocytes at the time of primary antigen stimulation Pgp-1 was previously shown to be a useful cell surface marker for distinguishing antigen-specific memory CD8+ T lymphocytes after immunization. Here we demonstrate that this observation extends to CD4+ T lymphocytes. Antigen-specific CD4+ T cells in mice immunized with sperm whale myoglobin or keyhole limpet hemocyanin were contained nearly exclusively in the minor Pgp-1+ subset.

    View details for Web of Science ID A1989T962500019

    View details for PubMedID 2564418

  • ENHANCED CELL-MEDIATED PROTECTION AGAINST FATAL ESCHERICHIA-COLI SEPTICEMIA INDUCED BY TREATMENT WITH RECOMBINANT IL-2 JOURNAL OF IMMUNOLOGY GORONZY, J., WEYAND, C., Quan, J., Fathman, C. G., OHANLEY, P. 1989; 142 (4): 1134-1138

    Abstract

    Administration of rIL-2 to BALB/c mice induces a rapid, cell-mediated response that is sufficient to protect mice from a lethal i.p. dose of Escherichia coli. Mice were protected from septic death if IL-2 was administered i.p. within 1 h after the bacterial challenge. Optimal protection was provided by treating the lethally challenged mice with rIL-2 at 1 and 5 h or 1, 5, and 10 h after the bacterial challenge and was dose-dependent (greater than or equal to 5.0 x 10(5) U/kg). Furthermore, treatment of mice with anti-IL-2R antibody abolished the protective effect induced by rIL-2 administration. These data suggest that the rIL-2-induced protection against septic death in mice is mediated by a cell type expressing a functional IL-2R. One potentially important therapeutic application of rIL-2 may be to modulate the course of sepsis once the host has been exposed to potentially lethal microbial pathogens.

    View details for Web of Science ID A1989T159900011

    View details for PubMedID 2644349

  • A NEW LOOK AT THE SHARED EPITOPE HYPOTHESIS AMERICAN JOURNAL OF MEDICINE Morel, P. A., Erlich, H. A., Fathman, C. G. 1988; 85 (6A): 20-22

    Abstract

    The striking correlation observed between T cell recognition and the sharing of the DR-beta-1 gene sequences (position 67-74) among patients with rheumatoid arthritis studied suggests that the third hypervariable region might be an important contribution to one restriction site for the putative causative agent(s) in rheumatoid arthritis. The fact that this sequence was found in DR1, DR4,Dw14, and DR4,Dw15 beta-1 genes lends support to the hypothesis that, in some cases, human leukocyte antigen and disease association may involve the association of discrete disease-related epitopes rather than entire human leukocyte antigen genes and that these epitopes are immunologically relevant in terms of T cell recognition. The association of these polymorphisms with susceptibility to rheumatoid arthritis would then support the hypothesis that binding and presentation of "arthritogenic peptides" by major histocompatibility complex class II molecules is one of the pathogenic events in developing rheumatoid arthritis.

    View details for Web of Science ID A1988R586200008

    View details for PubMedID 2462347

  • USE OF ANTI-L3T4 AND ANTI-IA TREATMENTS FOR PROLONGATION OF XENOGENEIC ISLET TRANSPLANTS TRANSPLANTATION Kaufman, D. S., Kong, C. S., Shizuru, J. A., Gregory, A. K., Fathman, C. G. 1988; 46 (2): 210-215

    Abstract

    The effects of T helper lymphocyte and Ia+ cell depletion were examined for their ability to independently and synergistically achieve prolongation of xenogeneic (rat-to-mouse) islet transplants. Recipient mice were depleted of T helper lymphocytes by short-term treatment with the anti-L3T4 monoclonal antibody GK1.5. Donor rat islets were treated prior to transplantation with a concentration of anti-Ia immunotoxin (13.4 x RT) that selectively depleted Ia+ cells within the islets while leaving functional insulin-secreting beta-cells unaffected. Anti-L3T4 treatment alone allowed transplants to be prolonged compared with untreated controls; however, all such treated mice rejected their xenogeneic transplant within 22 days. Although 13.4 x RT treatment of donor islets alone did not prolong engraftment, when donor rat islets were pretreated with the anti-Ia immunotoxin and grafted into L3T4-depleted mice, normoglycemia was maintained for greater than 50 days in 56% of transplants. These results suggest that neither L3T4 depletion nor anti-Ia immunotoxin treatment alone is enough to achieve indefinite survival of xenogeneic islets. However, decreasing the immunogenicity of the transplanted islets by anti-Ia immunotoxin treatment prior to transplantation into anti-L3T4 treated mice can allow greatly prolonged xenogeneic graft survival.

    View details for Web of Science ID A1988P695400005

    View details for PubMedID 2970132

  • IMMUNOTHERAPY OF THE NONOBESE DIABETIC MOUSE - TREATMENT WITH AN ANTIBODY TO T-HELPER LYMPHOCYTES SCIENCE Shizuru, J. A., TAYLOREDWARDS, C., Banks, B. A., Gregory, A. K., Fathman, C. G. 1988; 240 (4852): 659-662

    Abstract

    Spontaneous diabetes mellitus was blocked in nonobese diabetic mice by treatment with a monoclonal antibody against the L3T4 determinant present on the surface of T-helper lymphocytes. Sustained treatment with the monoclonal antibody led to cessation of the lymphocytic infiltration associated with the destruction of the insulin-producing beta cells. Moreover, the mice remained normoglycemic after the antibody therapy was stopped. These studies indicate that immunotherapy with monoclonal antibodies to the lymphocyte subset may not only halt the progression of diabetes, but may lead to long-term reversal of the disease after therapy has ended.

    View details for Web of Science ID A1988N126500035

    View details for PubMedID 2966437

  • ADMINISTRATION INVIVO OF RECOMBINANT INTERLEUKIN-2 PROTECTS MICE AGAINST SEPTIC DEATH JOURNAL OF CLINICAL INVESTIGATION WEYAND, C., GORONZY, J., Fathman, C. G., OHANLEY, P. 1987; 79 (6): 1756-1763

    Abstract

    Administration in vivo of recombinant interleukin 2 (rIL-2) to mice induces a polyclonal IgM response. When co-administered with a specific antigen, rIL-2 can enhance concentrations of murine IgM antibodies specific for the antigen by fivefold within 7 d of initial treatment. IgM antibodies that are induced after injection of rIL-2 include antibodies specific for J5, a cell wall core lipopolysaccharide (LPS) antigen that is shared by the different members of the Enterobactericeae family. We report here that mice pretreated with rIL-2 or immunized with J5 antigen 7 d before bacterial challenge were protected from septic death that is caused by intraperitoneal challenges with Escherichia coli. Optimal protection was provided by a combined J5 antigen and rIL-2 treatment. Acquisition of the rIL-2 and J5 antigen-induced protection against lethal bacterial infection coincided temporally with maximal serum IgM titers that also contained IgM antibodies specific for the J5 antigen. In passive immunization experiments, the affinity-purified IgM fraction in sera of rIL-2-treated animals was identified as necessary and sufficient for protection. The IgM-depleted serum had no protective effect. The nonspecific augmentation of host-defense mechanisms without the induction of endotoxin manifestations makes rIL-2 a potential candidate to any alternative LPS-containing vaccines for the prevention of bacterial infections by gram-negative organisms since the core LPS antigen is shared among gram-negative bacteria.

    View details for Web of Science ID A1987H575600029

    View details for PubMedID 3294901

  • MODULATION OF CD4 BY ANTIGENIC ACTIVATION JOURNAL OF IMMUNOLOGY Weyand, C. M., GORONZY, J., Fathman, C. G. 1987; 138 (5): 1351-1354

    Abstract

    Cell surface expression of CD4, an invariant membrane glycoprotein, is characteristic of the MHC class II-restricted T cell helper/inducer subset. Although the specificity and restriction patterns of T lymphocytes are determined by the T cell receptor for antigen, CD4 might represent an additional "interaction" molecule that is required to strengthen the interaction between T cells, antigen, and antigen-presenting cells. In this manuscript, we have shown that the cell surface expression of CD4 is correlated with activation of T cells. Data presented in this paper have demonstrated, for the first time, that antigenic stimulation of human T cell clones caused a decrease in the expression of the CD4 marker (as well as to the CD3 marker) to about 50% of the constitutive level. As previously demonstrated, PMA caused modulation of CD4 and CD3, which suggested that phosphorylation by protein kinase C played a crucial role in the regulation of the expression of both markers. The parallel down-regulation of CD3 and CD4 after antigen stimulation suggested that both markers might be members of a multimolecular complex mediating T cell activation.

    View details for Web of Science ID A1987G268700007

    View details for PubMedID 3100638

  • CLONING OF HUMAN ALLOREACTIVE T-CELLS METHODS IN ENZYMOLOGY GORONZY, J., WEYAND, C., Fathman, C. G. 1987; 150: 333-341

    View details for Web of Science ID A1987M168400028

    View details for PubMedID 3501522

  • LONG-TERM HUMORAL UNRESPONSIVENESS INVIVO, INDUCED BY TREATMENT WITH MONOCLONAL-ANTIBODY AGAINST L3T4 JOURNAL OF EXPERIMENTAL MEDICINE GORONZY, J., Weyand, C. M., Fathman, C. G. 1986; 164 (3): 911-925

    Abstract

    mAbs directed against the L3T4 molecule administered in vivo caused a severe and long lasting helper cell depletion in mice. Regeneration of the L3T4+ subpopulation occurred gradually (2-3 mo) after a single antibody treatment. Experiments were designed to examine the humoral immunocompetence of such anti-L3T4-treated animals during and after regeneration of the L3T4+ T cell subset. The animals were injected with anti-L3T4, immunized with soluble antigen, and challenged with antigen every 2 wk. Antibody responses to two antigens, sperm whale myoglobin (SpWMb) and KLH, which differ with regard to their immunogenicity, were compared. The lack of humoral immune responsiveness to either of these two antigens shorty after anti-L3T4 treatment responsiveness to either of these two antigens shortly after anti-L3T4 treatment was probably due to clonal depletion. The anti-L3T4-induced immunosuppressive effect on antibody production seemed to be determined in part by the preexisting T cell repertoire, as was suggested by the recovery of responsiveness to the highly immunogenic antigen KLH and the transient inhibitory effect of anti-L3T4 treatment in primed animals. The regenerating L3T4+ T cell subpopulation was relatively incompetent in initiating B cell responses. More than 40% of the L3T4+ T cell compartment had to recover to provide help for the production of anti-KLH antibodies, whereas elimination of 90% of the L3T4+ helper cells did not inhibit a primary anti-KLH response. Evidence for a heterogeneous composition of the L3T4+ subset came from experiments using rIL-2 in vivo. The addition of rIL-2 during early helper cell depletion improved the recovery of the humoral responsiveness without apparently affecting the kinetics of the regeneration of L3T4+ T cells. Interestingly, humoral unresponsiveness to the weakly immunogenic antigen SpWMb persisted for at least 120 d. This long lasting unresponsiveness could not be explained by clonal depletion, and suggested as one possibility that the presence of antigen during regeneration of the L3T4+ helper cell population may have influenced the ultimate T cell repertoire.

    View details for Web of Science ID A1986D904900017

    View details for PubMedID 3091757

  • GENE CONVERSION - A MECHANISM TO EXPLAIN HLA-D REGION AND DISEASE ASSOCIATION ANNALS OF THE NEW YORK ACADEMY OF SCIENCES Fathman, C. G., GORONZY, J., WEYAND, C. 1986; 475: 24-31

    Abstract

    In speculating about mechanisms that might give rise to T-cell epitopes appearing within different HLA-DR frameworks, we return to the hypothesis expressed above that suggests that gene-conversion-like events might be involved in shuffling the hypervariable segments of HLA-D region exons into alternative HLA-D region frameworks where they will still be recognized by the T cell (but not typed by conventional serology or mixed lymphocyte typing) as the "disease associated" HLA product. This might well explain the lack of stringent association between rheumatoid arthritis and HLA-DR4. It is possible, through the use of such alloreactive T-cell clones, that we might eventually define subgroups based upon presumed genetic susceptibility markers, which might allow therapeutic or prognostic assignment of patients with seropositive rheumatoid arthritis.

    View details for Web of Science ID A1986D835500003

    View details for PubMedID 2431644

  • SHARED T-CELL RECOGNITION SITES ON HUMAN HISTOCOMPATIBILITY LEUKOCYTE ANTIGEN CLASS-II MOLECULES OF PATIENTS WITH SEROPOSITIVE RHEUMATOID-ARTHRITIS JOURNAL OF CLINICAL INVESTIGATION GORONZY, J., Weyand, C. M., Fathman, C. G. 1986; 77 (3): 1042-1049

    Abstract

    Seropositive rheumatoid arthritis (RA) in adult and juvenile patients is associated with the serologic marker HLA-DR4. This association is incomplete; about one-third of the patients lack the disease-associated HLA-DR4 haplotype. The main biological function of class II molecules is to restrict the recognition of antigen by T lymphocytes. We therefore tested the hypothesis that patients with seropositive RA share T cell recognition sites for an unknown antigen and that such T cell "epitopes" are not identified by conventional serologic typing. We generated alloreactive human T cell clones by stimulating peripheral blood lymphocytes of normal donors against a lymphoblastoid cell line from a juvenile patient with seropositive RA. A panel of clones that recognized only HLA-Dw14 cells on a panel of homozygous typing cells was used to analyze class II molecules of adult patients with seropositive RA. By inhibition studies using monoclonal antibodies, the epitopes recognized by the different clones could be further characterized and assigned either to DR- or to DQ-encoded cell surface products. By using four different clones, it was possible to identify Dw14-associated T cell epitopes on all seropositive rheumatoid patients tested who typed HLA-DR4-positive and also on all eight DR4-negative patients tested. Approximately one-half of nonrheumatoid DR4-positive donors carried one or more determinants recognized by these clones; the expression of these allodeterminants in DR4-negative nonrheumatoid patients was rare (less than 10%). Thus, alloreactive human T cell clones are powerful tools to define T cell recognition sites on class II molecules that are not identified by conventional typing. Using T cell clones with specificities for determinants expressed on Dw14 homozygous typing lines, we were able to demonstrate shared epitopes on cells of all patients tested with seropositive RA irrespective of their HLA-D or HLA-DR type. These data suggest that major histocompatibility complex class II antigens of RA patients might be much more homogeneous than demonstrated by the incomplete HLA-DR4 association.

    View details for Web of Science ID A1986A542900051

    View details for PubMedID 2419361

  • HUMAN T-CELL CLONES USED TO DEFINE FUNCTIONAL EPITOPES ON HLA CLASS-II MOLECULES PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Weyand, C. M., GORONZY, J., Fathman, C. G. 1986; 83 (3): 762-766

    Abstract

    Polyclonal reagents have been used to define HLA class II molecules in conventional serologic and cellular typing. We generated human alloreactive T-cell clones to analyze the functional fine specificities of HLA class II molecules that might be important for the phenomenon of HLA and disease association. We chose to examine HLA-Dw14, an HLA-D specificity that has been associated with juvenile rheumatoid arthritis. In this paper we have presented data that suggest that conventional cellular typing does not reflect the distribution of T-cell epitopes on major histocompatibility complex class II molecules. We describe three alloreactive T-cell clones that have defined three separate Dw14-associated T-cell epitopes. Two of these epitopes were on a DR-region molecule; the third was located on a DQ-region product. In a panel of unrelated DR4-positive donors, these three DW14-associated determinants were present in a high frequency but were not linked to each other. Within the tested panel of DR4-positive cells, all possible combinatorial arrangements of these three allodeterminants were seen. The concurrent expression of any two of the three allodeterminants was equivalent to a positive typing response for Dw14. Our finding that HLA-Dw14 is not characterized by a unique allodeterminant but by the combinatorial recognition of independently distributed T-cell interaction sites suggests that analysis of HLA and disease association may be more clearly demonstrated through the use of human T-cell clones.

    View details for Web of Science ID A1986AZB0800050

    View details for PubMedID 2418442

  • HUMAN T-CELL CLONES AS DIAGNOSTIC-TOOLS AND POTENTIAL THERAPEUTIC REAGENTS IMMUNOLOGICAL INVESTIGATIONS WEYAND, C., GORONZY, J., Fathman, C. G. 1986; 15 (7): 591-624

    View details for Web of Science ID A1986F292100001

    View details for PubMedID 2433217

Conference Proceedings


  • Gene therapy for autoimmune disease Fathman, C. G., Costa, G. L., Seroogy, C. M. ACADEMIC PRESS INC ELSEVIER SCIENCE. 2000: S39-S43

    Abstract

    Autoantigen-specific CD4(+) T lymphocytes have been implicated in the pathogenesis of autoimmune diseases. Tissue-specific homing properties of autoantigen-specific CD4(+) T cells suggested that these cells might be ideal vehicles for delivery of retroviral-encoded regulatory proteins in a site-specific manner as a therapy for autoimmune diseases. Application of retroviral transduction of autoantigen-reactive CD4(+) T cells in gene therapy of autoimmunity must include systems capable of targeting these rare populations of antigen-activated T cells. Studies discussed below suggest that retroviral transduction of autoantigen-specific murine CD4(+) T cells may provide a method to target and isolate nontransformed autoantigen-specific murine CD4(+) T cells and provide a rational approach to gene therapy in animal models of autoimmunity.

    View details for Web of Science ID 000086408700007

    View details for PubMedID 10729236

  • Rat pancreatic islet and skin xenograft survival in CD4 and CD8 knockout mice Krieger, N. R., Ito, H., Fathman, C. G. ACADEMIC PRESS LTD ELSEVIER SCIENCE LTD. 1997: 309-315

    Abstract

    The relative contributions of the CD4+ and CD8+ T cell subpopulations in xenotransplant rejection were studied using CD4 and CD8 knockout (KO) mice. Wistar Furth (WF, RT1a) rat pancreatic islet or skin xenografts were transplanted into either CD4 or CD8 KO recipients and compared to wild-type controls. Long-term survival of WF islet xenografts was observed in the CD4 KO mice (MST, >66+/-8 days) whereas CD8 KO mice rejected their islet xenografts within 8 days, similar to controls (MST, 7+/-0.2 days). In contrast, WF skin xenografts were rejected in both CD4 and CD8 KO recipients within 8 days. CD4 KO recipients which maintained xenoislets >90 days posttransplant rejected WF skin grafts within 9 days, without rejecting their original islet xenografts. These results suggest that CD4+ cells are essential for mediating islet xenograft rejection. These data also suggest that the absence of either CD4+ or CD8+ T cells is not sufficient to prevent rejection of skin xenografts.

    View details for Web of Science ID A1997XG03300013

    View details for PubMedID 9218759

  • Immune regulation in type 1 diabetes Shimada, A., Charlton, B., Rohane, P., TAYLOREDWARDS, C., Fathman, C. G. ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD. 1996: 263-269

    Abstract

    The non-obese diabetic (NOD) mouse is an animal model of insulin-dependent diabetes mellitus (IDDM) that shows many of the characteristics of human IDDM. In the NOD model, there exists a discrepancy between the onset of insulitis and diabetes suggesting the potential existence of some form of immune regulation that delays beta cell destruction. Our transfer system using NOD-scid/scid (NOD-scid) mice as recipients of donor NOD cells suggested that immune regulatory cells exist in the periphery of NOD mice, not in the islets. These regulatory cells are considered to be memory CD4+ cells which show a Th2 (or Th zero) type cytokine profile following activation in vitro. The function of the memory CD4+ cells seems to change from protective to pathogenic as the disease progresses. Moreover, cytokine profiles of this CD4+ CD45RBlow (memory) population shifted from a Th2 (or Th zero) to a Th1 type response coincident with the onset of hyperglycaemia. These data suggest that the progression of NOD disease from insulitis to frank hyperglycaemia is under the control of CD4+ CD45RBlow immune 'regulatory' cells.

    View details for Web of Science ID A1996UL12400019

    View details for PubMedID 8738972

  • PEPTIDES AS THERAPY OF AUTOIMMUNE-DISEASE Fathman, C. G. JOHN WILEY & SONS LTD. 1993: 239-244

    View details for Web of Science ID A1993NN66400002

    View details for PubMedID 7924820

  • ANALYSIS OF THE TERNARY COMPLEX OF ANTIGEN, MHC AND T-CELL RECEPTOR - THE INFLUENCE OF THE T-CELL RECEPTOR V-BETA REPERTOIRE ON THE V-BETA GENE ELEMENT USAGE Ruberti, G., Livingstone, A., Danska, J. S., Gaur, A., Fathman, C. G. EDITIONS SCIENTIFIQUES ELSEVIER. 1991: 491-493

    View details for Web of Science ID A1991GL67400020

    View details for PubMedID 1721721

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