CD8+ T cells specific for the islet autoantigen IGRP are restricted in their T cell receptor chain usage

CD8+ T cells directed against beta cell autoantigens are considered relevant for the pathogenesis of type 1 diabetes. Using single cell T cell receptor sequencing of CD8+ T cells specific for the IGRP265-273 epitope, we examined whether there was expansion of clonotypes and sharing of T cell receptor chains in autoreactive CD8+ T cell repertoires. HLA-A*0201 positive type 1 diabetes patients (n = 19) and controls (n = 18) were analysed. TCR α- and β-chain sequences of 418 patient-derived IGRP265-273-multimer+ CD8+ T cells representing 48 clonotypes were obtained. Expanded populations of IGRP265-273-specific CD8+ T cells with dominant clonotypes that had TCR α-chains shared across patients were observed. The SGGSNYKLTF motif corresponding to TRAJ53 was contained in 384 (91.9%) cells, and in 20 (41.7%) patient-derived clonotypes. TRAJ53 together with TRAV29/DV5 was found in 15 (31.3%) clonotypes. Using next generation TCR α-chain sequencing, we found enrichment of one of these TCR α-chains in the memory CD8+ T cells of patients as compared to healthy controls. CD8+ T cell clones bearing the enriched motifs mediated antigen-specific target cell lysis. We provide the first evidence for restriction of T cell receptor motifs in the alpha chain of human CD8+ T cells with specificity to a beta cell antigen.

[1]  T. Wölfel,et al.  The use of HLA-A*0201-transfected K562 as standard antigen-presenting cells for CD8(+) T lymphocytes in IFN-gamma ELISPOT assays. , 2002, Journal of immunological methods.

[2]  E. Bonifacio,et al.  High Diversity in the TCR Repertoire of GAD65 Autoantigen-Specific Human CD4+ T Cells , 2015, The Journal of Immunology.

[3]  D. R. Gamble,et al.  In situ characterization of autoimmune phenomena and expression of HLA molecules in the pancreas in diabetic insulitis. , 1985, The New England journal of medicine.

[4]  A. Sewell,et al.  alphabeta T cell receptors as predictors of health and disease alphabeta T cell receptors as predictors of health and disease , 2017 .

[5]  E. Bonifacio,et al.  Vagaries of the ELISpot assay: specific detection of antigen responsive cells requires purified CD8(+) T cells and MHC class I expressing antigen presenting cell lines. , 2015, Clinical immunology.

[6]  Q. Ouyang,et al.  Recognition of HLA Class I–Restricted β-Cell Epitopes in Type 1 Diabetes , 2006, Diabetes.

[7]  F. Cucca,et al.  T-cell receptor repertoire analysis in monozygotic twins concordant and discordant for type 1 diabetes. , 2012, Immunobiology.

[8]  D. Price,et al.  A Single Autoimmune T Cell Receptor Recognizes More Than a Million Different Peptides* , 2011, The Journal of Biological Chemistry.

[9]  J. Drijfhout,et al.  Autoreactive CD8 T cells associated with β cell destruction in type 1 diabetes , 2005 .

[10]  Luigi Caimi,et al.  Identification of a public CDR3 motif and a biased utilization of T-cell receptor V beta and J beta chains in HLA-A2/Melan-A-specific T-cell clonotypes of melanoma patients , 2009, Journal of Translational Medicine.

[11]  Thorsten Meinl,et al.  KNIME - the Konstanz information miner: version 2.0 and beyond , 2009, SKDD.

[12]  J. K. Oeser,et al.  Deletion of the G6pc2 Gene Encoding the Islet-Specific Glucose-6-Phosphatase Catalytic Subunit–Related Protein Does Not Affect the Progression or Incidence of Type 1 Diabetes in NOD/ShiLtJ Mice , 2011, Diabetes.

[13]  Jérôme Lane,et al.  IMGT®, the international ImMunoGeneTics information system® , 2004, Nucleic Acids Res..

[14]  M. Atkinson,et al.  Demonstration of islet-autoreactive CD8 T cells in insulitic lesions from recent onset and long-term type 1 diabetes patients , 2012, The Journal of experimental medicine.

[15]  J. Drijfhout,et al.  Islet-Specific CTL Cloned from a Type 1 Diabetes Patient Cause Beta-Cell Destruction after Engraftment into HLA-A2 Transgenic NOD/SCID/IL2RG Null Mice , 2012, PloS one.

[16]  S. Tafuro,et al.  Progression of autoimmune diabetes driven by avidity maturation of a T-cell population , 2000, Nature.

[17]  Q. Ouyang,et al.  Recognition of HLA class I-restricted beta-cell epitopes in type 1 diabetes. , 2006, Diabetes.

[18]  D. Wegmann,et al.  Epitope specificity, cytokine production profile and diabetogenic activity of insulin‐specific T cell clones isolated from NOD mice , 1995, European journal of immunology.

[19]  E. Bonifacio,et al.  Measuring T cell receptor and T cell gene expression diversity in antigen-responsive human CD4+ T cells. , 2013, Journal of immunological methods.

[20]  M. Peakman,et al.  Translational Mini‐Review Series on Type 1 Diabetes:
Systematic analysis of T cell epitopes in autoimmune diabetes , 2007, Clinical and experimental immunology.

[21]  J. Bell,et al.  Human HLA-A0201-restricted cytotoxic T lymphocyte recognition of influenza A is dominated by T cells bearing the V beta 17 gene segment , 1995, The Journal of experimental medicine.

[22]  David I Stuart,et al.  A structural basis for immunodominant human T cell receptor recognition , 2003, Nature Immunology.

[23]  J. Drijfhout,et al.  CTLs are targeted to kill beta cells in patients with type 1 diabetes through recognition of a glucose-regulated preproinsulin epitope. , 2008, The Journal of clinical investigation.

[24]  F. Wong,et al.  Current approaches to measuring human islet‐antigen specific T cell function in type 1 diabetes , 2010, Clinical and experimental immunology.

[25]  Beta-cell autoimmunity. , 2012, Methods in molecular biology.

[26]  P. Doherty,et al.  Molecular basis for universal HLA-A*0201–restricted CD8+ T-cell immunity against influenza viruses , 2016, Proceedings of the National Academy of Sciences.

[27]  A. Sewell,et al.  αβ T cell receptors as predictors of health and disease , 2015, Cellular and Molecular Immunology.

[28]  Katherine K. Matthews,et al.  β-Cell–Specific CD8 T Cell Phenotype in Type 1 Diabetes Reflects Chronic Autoantigen Exposure , 2014, Diabetes.

[29]  E. Leiter,et al.  Major Histocompatibility Complex Class I-Deficient NOD-B2mnull Mice are Diabetes and Insulitis Resistant , 1994, Diabetes.

[30]  C. Janeway,et al.  CD8 T cell clones from young nonobese diabetic (NOD) islets can transfer rapid onset of diabetes in NOD mice in the absence of CD4 cells , 1996, The Journal of experimental medicine.

[31]  Thomas S. Watkins,et al.  Naive CD8+ T-cell precursors display structured TCR repertoires and composite antigen-driven selection dynamics , 2015, Immunology and cell biology.

[32]  P. van Endert,et al.  The Frequency and Immunodominance of Islet-Specific CD8+ T-cell Responses Change after Type 1 Diabetes Diagnosis and Treatment , 2008, Diabetes.

[33]  M. Dolz,et al.  Immunization of HLA Class I Transgenic Mice Identifies Autoantigenic Epitopes Eliciting Dominant Responses in Type 1 Diabetes Patients1 , 2007, The Journal of Immunology.

[34]  E. Naumova,et al.  Multiple Glycines in TCR α-Chains Determine Clonally Diverse Nature of Human T Cell Memory to Influenza A Virus1 , 2008, The Journal of Immunology.

[35]  Olga V. Britanova,et al.  Preparing Unbiased T-Cell Receptor and Antibody cDNA Libraries for the Deep Next Generation Sequencing Profiling , 2013, Front. Immunol..

[36]  D. Harlan,et al.  Analysis of self-antigen specificity of islet-infiltrating T cells from human donors with type 1 diabetes , 2016, Nature Medicine.

[37]  K. Kedzierska,et al.  Proinsulin-Specific, HLA-DQ8, and HLA-DQ8-Transdimer–Restricted CD4+ T Cells Infiltrate Islets in Type 1 Diabetes , 2014, Diabetes.

[38]  E. Bonifacio,et al.  Abundant cytomegalovirus (CMV) reactive clonotypes in the CD8+ T cell receptor alpha repertoire following allogeneic transplantation , 2016, Clinical and experimental immunology.

[39]  Irene Jarchum,et al.  Identification of novel IGRP epitopes targeted in type 1 diabetes patients. , 2008, Clinical immunology.

[40]  C. Boitard,et al.  Recognition of a subregion of human proinsulin by class I-restricted T cells in type 1 diabetic patients. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[41]  P. van Endert,et al.  ZnT8 Is a Major CD8+ T Cell–Recognized Autoantigen in Pediatric Type 1 Diabetes , 2012, Diabetes.

[42]  J. Drijfhout,et al.  Autoreactive CD8 T cells associated with beta cell destruction in type 1 diabetes. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[43]  D. Finegood,et al.  Prediction of spontaneous autoimmune diabetes in NOD mice by quantification of autoreactive T cells in peripheral blood. , 2003, The Journal of clinical investigation.

[44]  E. Huseby,et al.  Indoctrinating T cells to attack pathogens through homeschooling. , 2015, Trends in immunology.

[45]  G. Eisenbarth,et al.  T cell receptor restriction of diabetogenic autoimmune NOD T cells. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[46]  J. Shabanowitz,et al.  Identification of the β cell antigen targeted by a prevalent population of pathogenic CD8+ T cells in autoimmune diabetes , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[47]  Y. Naumov,et al.  Narrowing of Human Influenza A Virus-Specific T Cell Receptor α and β Repertoires with Increasing Age , 2015, Journal of Virology.

[48]  H. Thomas,et al.  Functional cytotoxic T lymphocytes against IGRP206‐214 predict diabetes in the non‐obese diabetic mouse , 2014, Immunology and cell biology.

[49]  Bart O. Roep,et al.  Simultaneous Detection of Circulating Autoreactive CD8+ T-Cells Specific for Different Islet Cell–Associated Epitopes Using Combinatorial MHC Multimers , 2010, Diabetes.

[50]  G. Eisenbarth,et al.  Dual overlapping peptides recognized by insulin peptide B:9-23 T cell receptor AV13S3 T cell clones of the NOD mouse. , 2000, Journal of autoimmunity.

[51]  C. Farina,et al.  Cytotoxic T-lymphocyte clones from different patients display limited T-cell-receptor variable-region gene usage in HLA-A2-restricted recognition of the melanoma antigen Melan-A/MART-1. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[52]  D. Roopenian,et al.  Major histocompatibility complex class I-restricted T cells are required for all but the end stages of diabetes development in nonobese diabetic mice and use a prevalent T cell receptor alpha chain gene rearrangement. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[53]  A. Sewell,et al.  Hotspot autoimmune T cell receptor binding underlies pathogen and insulin peptide cross-reactivity , 2016, The Journal of clinical investigation.

[54]  M. Dolz,et al.  CD8+ T-Cell Responses Identify β-Cell Autoimmunity in Human Type 1 Diabetes , 2007, Diabetes.

[55]  T. Utsugi,et al.  Beta-cell-cytotoxic CD8+ T cells from nonobese diabetic mice use highly homologous T cell receptor alpha-chain CDR3 sequences. , 1995, Journal of immunology.