Detection of GAD65-specific T-cells by major histocompatibility complex class II tetramers in type 1 diabetic patients and at-risk subjects.
Soluble HLA-DR401 or -DR404 tetramers containing a peptide corresponding to an immunodominant epitope from human GAD65 were used to analyze peripheral blood T-cells of newly diagnosed type 1 diabetic patients and at-risk subjects. Peripheral blood mononuclear cells were expanded on antigen-presenting cells presenting GAD65 peptide and subsequently activated with specific plate-bound class II-peptide monomers. T-cell activation defined in flow cytometry by CD4(high) and/or CD25 markers were observed in all type 1 diabetic patients and some at-risk subjects, but not in normal control subjects. The activated T-cells stained positive with tetramers containing the GAD65 epitope 555-567. Tetramer-positive cells were CD4(high) T-cells with high avidity for an immunodominant GAD65 T-cell epitope. Phenotyping of T-cells utilizing HLA class II tetramers provides a new tool to characterize the autoimmune response in type 1 diabetes.
Reijonen
Helena
H
Virginia Mason Research Center, Benaroya Research Institute, Seattle, Washington 98101-2795, USA. reijonen@vmresearch.org
Novak
Erik J
EJ
Kochik
Sharon
S
Heninger
Anne
A
Liu
Andrew W
AW
Kwok
William W
WW
Nepom
Gerald T
GT
D000293:Adolescent; D000328:Adult; D003922:Diabetes Mellitus, Type 1 / Q000453:epidemiology / Q000276:immunology*; D000939:Epitopes / Q000276:immunology / Q000494:pharmacology; D005434:Flow Cytometry; D005968:Glutamate Decarboxylase / Q000276:immunology* / Q000494:pharmacology; D015804:HLA-DR4 Antigen / Q000276:immunology*; D006801:Humans; D016130:Immunophenotyping; D066298:In Vitro Techniques; D007527:Isoenzymes / Q000276:immunology* / Q000494:pharmacology; D008213:Lymphocyte Activation / Q000187:drug effects / Q000276:immunology; D012307:Risk Factors; D013601:T-Lymphocytes / Q000276:immunology*
D000939:Epitopes; D015804:HLA-DR4 Antigen; D007527:Isoenzymes; D005968:Glutamate Decarboxylase; C401141:glutamate decarboxylase 2