Flow cytometry for the analysis of T cells expressing CD69 after stimulation with glutamic acid decarboxylase.
Type 1 diabetes mellitus is a T-cell mediated autoimmune disease in which the insulin-producing pancreatic beta cells are selectively destroyed. We recently found that the detection of cell-mediated immune response to glutamic acid decarboxylase (GAD) was more useful than the detection of specific autoantibodies for the diagnosis of type 1 diabetes mellitus. In this study, we established a flow cytometric analysis for the detection of activated T cells in whole venous blood, obtained from diabetic patients and normal controls after stimulation by GAD. Two millitiers of peripheral venous blood and 6 hours incubation time were used for performing the test. It was found that 33% (3/9) type 1 diabetic patients, 7.7% (1/13) type 2 diabetic patients and neither patients with fibrocalculous pancreatopathy nor normal controls had > or = 20% CD8+ T cells expressing CD69. The results suggest that flow cytometry may be a useful tool for the detection of surrogate markers of type 1 diabetes mellitus.
Boonyasrisawat
Watip
W
Department of Immunology, Faculty of Medicine, Siriraj Hospital Mahidol University, Bangkok, Thailand.
Banchuin
Napatawn
N
Pattanapanyasat
Kovit
K
Deerochanawong
Chaicharn
C
Yenchitsomanus
Pa-Thai
PT
Ploybutr
Sirirat
S
Vannasaeng
Sathit
S
D000293:Adolescent; D000328:Adult; D000368:Aged; D015703:Antigens, CD / Q000096:biosynthesis* / Q000276:immunology*; D000945:Antigens, Differentiation, T-Lymphocyte / Q000096:biosynthesis* / Q000276:immunology* / Q000378:metabolism; D003922:Diabetes Mellitus, Type 1 / Q000276:immunology* / Q000378:metabolism*; D003924:Diabetes Mellitus, Type 2 / Q000276:immunology* / Q000378:metabolism*; D004306:Dose-Response Relationship, Immunologic; D005260:Female; D005434:Flow Cytometry; D005968:Glutamate Decarboxylase / Q000096:biosynthesis* / Q000276:immunology*; D006801:Humans; D007111:Immunity, Cellular / Q000276:immunology; D037181:Lectins, C-Type; D008213:Lymphocyte Activation / Q000187:drug effects; D008297:Male; D008875:Middle Aged; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism*; D013785:Thailand; D000099192:CD69 Antigens
D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D005968:Glutamate Decarboxylase; D037181:Lectins, C-Type; D000099192:CD69 Antigens