Autoreactive T cell responses show proinflammatory polarization in diabetes but a regulatory phenotype in health.

According to the quality of response they mediate, autoreactive T cells recognizing islet beta cell peptides could represent both disease effectors in the development of type 1 diabetes (T1DM) and directors of tolerance in nondiabetic individuals or those undergoing preventative immunotherapy. A combination of the rarity of these cells, inadequate technology, and poorly defined epitopes, however, has hampered examination of this paradigm. We have identified a panel of naturally processed islet epitopes by direct elution from APCs bearing HLA-DR4. Employing these epitopes in a sensitive, novel cytokine enzyme-linked immunosorbent spot assay, we show that the quality of autoreactive T cells in patients with T1DM exhibits extreme polarization toward a proinflammatory Th1 phenotype. Furthermore, we demonstrate that rather than being unresponsive, the majority of nondiabetic, HLA-matched control subjects also manifest a response against islet peptides, but one that shows extreme T regulatory cell (Treg, IL-10-secreting) bias. We conclude that development of T1DM depends on the balance of autoreactive Th1 and Treg cells, which may be open to favorable manipulation by immune intervention.

[1]  K. Elsegood,et al.  Humoral and cellular immune responses to proinsulin in adults with newly diagnosed type 1 diabetes , 2003, Diabetes/metabolism research and reviews.

[2]  M. Peakman,et al.  Promiscuous binding of proinsulin peptides to Type 1 diabetes-permissive and -protective HLA class II molecules , 2003, Diabetologia.

[3]  P. Ott,et al.  Cytokine detection by ELISPOT: relevance for immunological studies in type 1 diabetes , 2002, Diabetes/metabolism research and reviews.

[4]  R. Coffman,et al.  Interleukin-10 and the interleukin-10 receptor. , 2001, Annual review of immunology.

[5]  P. Bingley,et al.  Update on Major Trials for the Prevention of Type I Diabetes Mellitus: The American Diabetes Prevention Trial (DPT-1) and the European Nicotinamide Diabetes Intervention Trial (ENDIT) , 2001, Journal of pediatric endocrinology & metabolism : JPEM.

[6]  C Benoist,et al.  T helper cell subsets in insulin-dependent diabetes. , 1995, Science.

[7]  B. Roep.  T-Cell Responses to Autoantigens in IDDM: The Search for the Holy Grail , 1996, Diabetes.

[8]  E. Keeler,et al.  Primer on certain elements of medical decision making. , 1975, The New England journal of medicine.

[9]  M. Peakman,et al.  T-Cell markers in Type I diabetes: progress, prospects and realistic expectations , 2000, Diabetologia.

[10]  D. J. Cowley,et al.  Expression, purification and characterization of recombinant human proinsulin , 1997, FEBS letters.

[11]  A. Abbas,et al.  The enemy within: keeping self-reactive T cells at bay in the periphery , 2002, Nature Reviews Immunology.

[12]  P. Bingley,et al.  A comparison of the prevalence of islet autoantibodies in children from two countries with differing incidence of diabetes , 2001, Diabetologia.

[13]  P. Bingley,et al.  Optimized autoantibody-based risk assessment in family members. Implications for future intervention trials. , 1999, Diabetes care.

[14]  P. Ott,et al.  Comparison of cytokine ELISpot assay formats for the detection of islet antigen autoreactive T cells. Report of the third immunology of diabetes society T-cell workshop. , 2003, Journal of autoimmunity.

[15]  B. Boehm,et al.  Predominantly recognized proinsulin T helper cell epitopes in individuals with and without islet cell autoimmunity. , 2002, Journal of autoimmunity.

[16]  Y. Levo,et al.  Cutting Edge: Anti-Inflammatory Properties of Low Levels of IFN-γ1 , 2002, The Journal of Immunology.

[17]  M. Peakman,et al.  Enhanced T cell proliferation and increased responder frequency following delivery of antigen to the antigen-presenting cell; B cell dependency and use in detection of autoreactive T cells. , 1998, Journal of immunological methods.

[18]  H. Mcdevitt,et al.  Th1 and Th2 CD4+ T cells in the pathogenesis of organ-specific autoimmune diseases. , 1995, Immunology today.

[19]  P. van Endert,et al.  Autoreactive T cell responses in insulin-dependent (Type 1) diabetes mellitus. Report of the first international workshop for standardization of T cell assays. , 1999, Journal of autoimmunity.

[20]  S. Azar,et al.  T Helper Type 1 and 2 Cytokines Mediate the Onset and Progression of Type I (Insulin-Dependent) Diabetes , 1999 .

[21]  I. Cohen,et al.  β-cell function in new-onset type 1 diabetes and immunomodulation with a heat-shock protein peptide (DiaPep277): a randomised, double-blind, phase II trial , 2001, The Lancet.

[22]  S. Azar,et al.  Clinical review 103: T helper type 1 and 2 cytokines mediate the onset and progression of type I (insulin-dependent) diabetes. , 1999, The Journal of clinical endocrinology and metabolism.

[23]  J. Bluestone,et al.  Suppressor T cells – they’re back and critical for regulation of autoimmunity! , 2001, Immunological reviews.

[24]  Antonio Lanzavecchia,et al.  Universally immunogenic T cell epitopes: promiscuous binding to human MHC class II and promiscuous recognition by T cells , 1989, European journal of immunology.

[25]  R. Mackin.  Streamlined procedure for the production of normal and altered versions of recombinant human proinsulin. , 1999, Protein expression and purification.

[26]  I. Cohen,et al.  Beta-cell function in new-onset type 1 diabetes and immunomodulation with a heat-shock protein peptide (DiaPep277): a randomised, double-blind, phase II trial. , 2001, Lancet.

[27]  A. Tomlinson,et al.  Naturally processed and presented epitopes of the islet cell autoantigen IA-2 eluted from HLA-DR4. , 1999, The Journal of clinical investigation.

[28]  J. She.  Susceptibility to type I diabetes: HLA-DQ and DR revisited. , 1996, Immunology today.

[29]  B. Walker,et al.  Ex vivo analysis of human memory CD4 T cells specific for hepatitis C virus using MHC class II tetramers. , 2003, The Journal of clinical investigation.

[30]  W. Suarez-Pinzon,et al.  Th1 to Th2 Cytokine Shifts in Nonobese Diabetic Mice: Sometimes an Outcome, Rather Than the Cause, of Diabetes Resistance Elicited by Immunostimulation1 , 2001, The Journal of Immunology.

[31]  S. Virgiliis,et al.  T cell epitopes of insulin defined in HLA-DR4 transgenic mice are derived from preproinsulin and proinsulin. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[32]  P. Bingley,et al.  Diabetes Antibody Standardization Program: first assay proficiency evaluation. , 2003, Diabetes.

[33]  H. Rammensee,et al.  SYFPEITHI: database for MHC ligands and peptide motifs , 1999, Immunogenetics.

[34]  G. Trinchieri.  Regulatory Role of T Cells Producing both Interferon γ and Interleukin 10 in Persistent Infection , 2001, The Journal of experimental medicine.

[35]  M. Atkinson,et al.  Type 1 diabetes: new perspectives on disease pathogenesis and treatment , 2001, The Lancet.

[36]  J R Beck,et al.  The use of relative operating characteristic (ROC) curves in test performance evaluation. , 1986, Archives of pathology & laboratory medicine.

[37]  J. Killestein.  Anti-CD3 monoclonal antibody in new-onset type 1 diabetes mellitus. , 2002, The New England journal of medicine.

[38]  W. Kwok,et al.  HLA Class II-Restricted CD4+ T Cell Responses Directed Against Influenza Viral Antigens Postinfluenza Vaccination 1 , 2003, The Journal of Immunology.