title: 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.
abstract: 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.
authors: INSERM U986, DeAR Lab Avenir, Saint Vincent de Paul Hospital, and Paris Descartes University, Paris, France. Roberto.Mallone@inserm.fr
authors: R
authors: R
authors: Mallone
authors: M
authors: M
authors: Scotto
authors: C N
authors: CN
authors: Janicki
authors: E A
authors: EA
authors: James
authors: L
authors: L
authors: Fitzgerald-Miller
authors: R
authors: R
authors: Wagner
authors: P
authors: P
authors: Gottlieb
authors: J
authors: J
authors: Thorpe
authors: N
authors: N
authors: Jospe
authors: I
authors: I
authors: Durinovic-Bellò
authors: C
authors: C
authors: Boitard
authors: O
authors: O
authors: Lou
authors: C M
authors: CM
authors: Dayan
authors: F S
authors: FS
authors: Wong
mesh_terms: D000293:Adolescent; D000328:Adult; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D003922:Diabetes Mellitus, Type 1 / Q000276:immunology*; D058501:Enzyme-Linked Immunospot Assay; D018984:Epitopes, T-Lymphocyte / Q000276:immunology*; D005968:Glutamate Decarboxylase / Q000276:immunology; D015234:HLA-A Antigens / Q000276:immunology; D015395:Histocompatibility Antigens Class I / Q000276:immunology*; D006801:Humans; D007328:Insulin / Q000276:immunology*; D007371:Interferon-gamma / Q000276:immunology; D011498:Protein Precursors / Q000276:immunology*
chemical_list: D018984:Epitopes, T-Lymphocyte; D015234:HLA-A Antigens; D015395:Histocompatibility Antigens Class I; D007328:Insulin; D011498:Protein Precursors; C013737:preproinsulin; D007371:Interferon-gamma; D005968:Glutamate Decarboxylase