[en] Type 1 diabetes (T1D) is a T cell-mediated autoimmune disease in which the insulin-producing β cells within the pancreas are destroyed. Identification of target Ags and epitopes of the β cell-reactive T cells is important both for understanding T1D pathogenesis and for the rational development of Ag-specific immunotherapies for the disease. Several studies suggest that proinsulin is an early and integral target autoantigen in T1D. However, proinsulin epitopes recognized by human CD4+ T cells have not been comprehensively characterized. Using a dye dilution-based T cell cloning method, we generated and characterized 24 unique proinsulin-specific CD4+ T cell clones from the peripheral blood of 17 individuals who carry the high-risk DR3-DQ2 and/or DR4-DQ8 HLA class II haplotypes. Some of the clones recognized previously reported DR4-restricted epitopes within the C-peptide (C25-35) or A-chain (A1-15) of proinsulin. However, we also characterized DR3-restricted epitopes within both the B-chain (B16-27 and B22-C3) and C-peptide (C25-35). Moreover, we identified DQ2-restricted epitopes within the B-chain and several DQ2- or DQ8-restricted epitopes within the C-terminal region of C-peptide that partially overlap with previously reported DQ-restricted epitopes. Two of the DQ2-restricted epitopes, B18-26 and C22-33, were shown to be naturally processed from whole human proinsulin. Finally, we observed a higher frequency of CDR3 sequences matching the TCR sequences of the proinsulin-specific T cell clones in pancreatic lymph node samples compared with spleen samples. In conclusion, we confirmed several previously reported epitopes but also identified novel (to our knowledge) epitopes within proinsulin, which are presented by HLA class II molecules associated with T1D risk.
Disciplines :
Immunology & infectious disease
Author, co-author :
Ihantola, Emmi-Leena ; Department of Clinical Microbiology, Institute of Clinical Medicine, University of Eastern Finland, FI-70210 Kuopio, Finland
Ilmonen, Henna ; Department of Clinical Microbiology, Institute of Clinical Medicine, University of Eastern Finland, FI-70210 Kuopio, Finland
Kailaanmäki, Anssi ; Department of Clinical Microbiology, Institute of Clinical Medicine, University of Eastern Finland, FI-70210 Kuopio, Finland
Rytkönen-Nissinen, Marja ; Department of Clinical Microbiology, Institute of Clinical Medicine, University of Eastern Finland, FI-70210 Kuopio, Finland
Azam, Aurélien ; Commissariat à l'Energie Atomique et aux Energies Alternatives-Saclay, Université Paris-Saclay, Service d'Ingénierie Moléculaire des Protéines, 91191 Gif Sur Yvette, France
Maillère, Bernard; Commissariat à l'Energie Atomique et aux Energies Alternatives-Saclay, Université Paris-Saclay, Service d'Ingénierie Moléculaire des Protéines, 91191 Gif Sur Yvette, France
Lindestam Arlehamn, Cecilia S ; La Jolla Institute for Immunology, La Jolla, CA 92037
Sette, Alessandro; La Jolla Institute for Immunology, La Jolla, CA 92037 ; Department of Medicine, University of California San Diego, La Jolla, CA 92093
Motwani, Keshav ; Department of Pathology, Immunology and Laboratory Medicine, University of Florida Diabetes Institute, Gainesville, FL 32610
Seay, Howard R ; Department of Pathology, Immunology and Laboratory Medicine, University of Florida Diabetes Institute, Gainesville, FL 32610
Brusko, Todd M ; Department of Pathology, Immunology and Laboratory Medicine, University of Florida Diabetes Institute, Gainesville, FL 32610 ; Department of Pediatrics, University of Florida, College of Medicine Gainesville, FL 32610
Knip, Mikael ; Tampere Center for Child Health Research, Tampere University Hospital, FI-33520 Tampere, Finland ; Children's Hospital, University of Helsinki and Helsinki University Hospital, FI-00014 Helsinki, Finland ; Research Program for Clinical and Molecular Metabolism, Faculty of Medicine, University of Helsinki, FI-00014 Helsinki, Finland ; Folkhälsan Research Center, FI-00290 Helsinki, Finland
Veijola, Riitta ; PEDEGO Research Unit, Department of Pediatrics, Medical Research Center, Oulu University Hospital and University of Oulu, FI-90014 Oulu, Finland
Toppari, Jorma ; Department of Pediatrics, Turku University Hospital, FI-20521 Turku, Finland ; Institute of Biomedicine, Research Centre for Integrative Physiology and Pharmacology, University of Turku, FI-20520 Turku, Finland
Ilonen, Jorma ; Immunogenetics Laboratory, Institute of Biomedicine, University of Turku, FI-20520 Turku, Finland ; Clinical Microbiology, Turku University Hospital, FI-20521 Turku, Finland, and
Kinnunen, Tuure ; Department of Clinical Microbiology, Institute of Clinical Medicine, University of Eastern Finland, FI-70210 Kuopio, Finland, tuure.kinnunen@uef.fi ; Eastern Finland Laboratory Centre (ISLAB), FI-70210 Kuopio, Finland
This work was supported by the Academy of Finland (Decision 307320), the Sigrid Jusélius Foundation, State Research Funding (VTR), and the Finnish Diabetes Research Foundation. The Finnish Type 1 Diabetes Prediction and Prevention study was supported by the Academy of Finland (Decisions 250114 and 286765), the Sigrid Jusélius Foundation, and the Juvenile Diabetes Research Foundation International (JDRF). The Network for Pancreatic Organ Donors with Diabetes is supported by the JDRF, with cooperative mechanistic study support by The Leona M. and Harry B. Helmsley Charitable Trust (to T.M.B.). The funders had no role in the design and conduct of the study; in the collection, analysis, and interpretation of the data; and in the preparation, review, or approval of the manuscript.
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