Evidence for in vivo primed and expanded autoreactive T cells as a specific feature of patients with type 1 diabetes.
Identifying beta cell autoantigen-reactive T cells that are involved in the pathogenesis of type 1 diabetes has been troublesome for many laboratories. Disease-relevant autoreactive T cells should be in vivo Ag experienced. The aim of this study was to test this hypothesis and then use this principle as a strategy for identifying diabetes-relevant autoreactive T cells. In this study, a CSFE dilution assay was used to detect glutamic acid decarboxylase 65 (GAD65)- and insulin-responsive T cells and HLA-0201*-GAD65(114-122) pentamers were used to detect CD8(+) GAD-responsive T cells in memory CD45RO(+) and naive CD45RO(-) cell populations from patients with type 1 diabetes and healthy control subjects. T cell proliferative history was evaluated by flow cytometry telomere length measurement. CD4(+) and CD8(+) T cells specific for GAD65 and insulin were present in patients with type 1 diabetes and control subjects. Within the naive CD45RO(-) cells, CD4(+) and CD8(+) T cell responses were similar between patients and controls. Within the memory CD45RO(+) cells, CD4(+) T cell responses against whole GAD65 and insulin and HLA-0201*-GAD65(114-122) pentamer-positive CD8(+) T cells were found in patients with type 1 diabetes, but not in control subjects (p < 0.05 for all). Responding cells from the CD45RO(+) T cell population had substantially shorter telomere lengths than responding cells from the CD45RO(-) cell population. Diabetes-specific autoreactive T cells in the circulation have uniquely undergone sustained in vivo proliferation and differentiation into memory T cells. Prior selection of these cells is possible and is a way to identify diabetes-relevant target Ags and epitopes.
Monti
Paolo
P
Telethon-Juvenile Diabetes Research Foundation Center for Beta Cell Replacement, San Raffaele Scientific Institute, Milan, Italy.
Scirpoli
Miriam
M
Rigamonti
Andrea
A
Mayr
Anya
A
Jaeger
Annika
A
Bonfanti
Riccardo
R
Chiumello
Giuseppe
G
Ziegler
Anette G
AG
Bonifacio
Ezio
E
D000293:Adolescent; D000328:Adult; D001324:Autoantigens / Q000276:immunology*; D002454:Cell Differentiation / Q000276:immunology*; D002478:Cells, Cultured; D002648:Child; D003922:Diabetes Mellitus, Type 1 / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D005260:Female; D005452:Fluoresceins; D005968:Glutamate Decarboxylase / Q000378:metabolism; D006801:Humans; D007156:Immunologic Memory / Q000276:immunology; D007201:Indicator Dilution Techniques; D007328:Insulin / Q000378:metabolism; D017493:Leukocyte Common Antigens / Q000276:immunology; D008297:Male; D010641:Phenotype; D012307:Risk Factors; D013388:Succinimides; D013601:T-Lymphocytes / Q000166:cytology* / Q000276:immunology*; D016615:Telomere / Q000235:genetics
C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester; D001324:Autoantigens; D005452:Fluoresceins; D007328:Insulin; D013388:Succinimides; D017493:Leukocyte Common Antigens; D005968:Glutamate Decarboxylase; C401141:glutamate decarboxylase 2