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

Diabetes Metab Res Rev. 2011 Nov;27(8):720-6. 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; PPIA12-20 and PPIB10-18) 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.

Publication types

  • Multicenter Study
  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Adolescent
  • Adult
  • CD8-Positive T-Lymphocytes / immunology*
  • Diabetes Mellitus, Type 1 / immunology*
  • Enzyme-Linked Immunospot Assay
  • Epitopes, T-Lymphocyte / immunology*
  • Glutamate Decarboxylase / immunology
  • HLA-A Antigens / immunology
  • Histocompatibility Antigens Class I / immunology*
  • Humans
  • Insulin / immunology*
  • Interferon-gamma / immunology
  • Protein Precursors / immunology*


  • Epitopes, T-Lymphocyte
  • HLA-A Antigens
  • Histocompatibility Antigens Class I
  • Insulin
  • Protein Precursors
  • preproinsulin
  • Interferon-gamma
  • Glutamate Decarboxylase