Immunology of Diabetes Society T-Cell Workshop: HLA class I tetramer-directed epitope validation initiative T-Cell Workshop Report-HLA Class I Tetramer Validation Initiative

• Published in: Diabetes/metabolism research and reviews Vol. 27; no. 8; pp. 720 - 726
• Main Authors: Mallone, R., Scotto, M., Janicki, C. N., James, E. A., Fitzgerald-Miller, L., Wagner, R., Gottlieb, P., Thorpe, J., Jospe, N., Durinovic-Bellò, I., Boitard, C., Lou, O., Dayan, C. M., Wong, F. S.
• Format: Journal Article
• Language: English
• Published: Chichester, UK John Wiley & Sons, Ltd 01.11.2011; Wiley Subscription Services, Inc
• Subjects: Adolescent; Adult; alpha -Interferon; antigen; autoimmunity; CD8 antigen; CD8-Positive T-Lymphocytes - immunology; Conferences; Diabetes mellitus; Diabetes Mellitus, Type 1 - immunology; ELISpot; Enzyme-linked immunosorbent assay; Enzyme-Linked Immunospot Assay; Epitopes; Epitopes, T-Lymphocyte - immunology; GAD; Glutamate decarboxylase; Glutamate Decarboxylase - immunology; Histocompatibility antigen HLA; Histocompatibility Antigens Class I - immunology; HLA-A Antigens - immunology; Humans; insulin; Insulin - immunology; Interferon; Interferon-gamma - immunology; Lymphocytes T; Peripheral blood; Preproinsulin; Protein Precursors - immunology; Reviews
• ISSN: 1520-7552; 1520-7560; 1520-7560
• DOI: 10.1002/dmrr.1243

Background Identification of T‐cell reactivity to β‐cell antigen epitopes is an important goal for studying pathogenesis and for designing and monitoring of immunotherapeutic interventions in type 1 diabetes (T1D). Methods We performed a multicentre validation of known human leukocyte antigen (HLA) class I CD8+ T‐cell epitopes. To this end, peripheral blood T‐cell responses were measured in 35 recently (<2 years) diagnosed HLA‐A*02:01+ T1D patients using blind‐coded HLA‐A2 tetramers (TMrs) and pentamers (PMrs), encompassing two epitopes of preproinsulin (PPI; PPI$\def\endash{\hbox{--}} _{\rm{A12\endash 20}}$ and PPI$\def\endash{\hbox{--}} _{\rm{B10\endash 18}}$) and two epitopes of glutamic acid decarboxylase (GAD; GAD114–122 and GAD536–545). We also compared the readout of TMrs and PMrs with a CD8+ T‐cell interferon‐γ enzyme‐linked immunospot assay. Results Despite the minute frequencies of autoreactive cells detected by TMrs/PMrs, most (73–77%) T1D patients had responses to one or more of the epitopes used. All four epitopes were recognized by T1D patients, with a prevalence ranging from 5 to 25%. TMrs and PMrs detected more positive responses to the β‐cell epitopes than CD8+ T‐cell interferon‐γ enzyme‐linked immunospot. However, concordance between positive responses to TMrs and PMrs was limited. Conclusions Using a multicentre blind‐coded setup and three different T‐cell assays, we have validated PPI and GAD epitopes as commonly recognized CD8+ T‐cell targets in recently diagnosed T1D patients. Both TMrs and PMrs showed higher detection sensitivity than the CD8+ T‐cell interferon‐γ enzyme‐linked immunospot assay. However, there are some important methodological issues that need to be addressed in using these sensitive techniques for detecting low frequency responses. Copyright © 2011 John Wiley & Sons, Ltd.