Detection of GAD65-reactive T-Cells in type 1 diabetes by immunoglobulin-free ELISPOT assays

Kotani, Reiko, Nagata, Masao, Moriyama, Hiroaki, Nakayama, Maki, Yamada, Katsumi, Chowdhury, Shahead Ali, Chakrabarty, Sagarika, Jin, Zhenzi, Yasuda, Hisafumi, and Yokono, Koichi (2002) Detection of GAD65-reactive T-Cells in type 1 diabetes by immunoglobulin-free ELISPOT assays. Diabetes Care, 25 (8). pp. 1390-1397.

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Abstract

OBJECTIVE—To investigate the prevalence of ß-cell autoantigen-reactive peripheral T-cells in type 1 diabetes, we developed an immunoglobulin-free enzyme-linked immunospot (ELISPOT) assay and assessed its usefulness for diagnosing this disease.

RESEARCH DESIGN AND METHODS—Cellular immune responses to ß -cell autoantigens were studied both by immunoglobulin-free proliferation assays and ELISPOT assays in 33 patients with type 1 diabetes and 15 patients with type 2 diabetes, compared with 23 healthy control subjects. Autoantibodies against GAD65 and IA-2 were measured by radioimmunoassay.

RESULTS—Significant proliferative responses to GAD65 were observed in 10 of 31 (32.3%) type 1 diabetic patients (P < 0.05), whereas GAD65-reactive -interferon (IFN-)-secreting cells were detected in 22 of 33 patients (66.7%) by ELISPOT assay (P < 0.001). Of patients negative for both GAD65 and IA-2, five of six (83.3%) showed IFN- positivity in ELISPOT and two of five (40.0%) showed significant proliferation against GAD65.

CONCLUSIONS—Using a newly developed ELISPOT assay, GAD-reactive T-helper 1 cells in PBMC of type 1 diabetic patients could be identified at a higher frequency than by the proliferation assay. Therefore, the immunoglobulin-free ELISPOT assay is an excellent tool for detecting T-cell reactivity to autoantigens with greater specificity and, in combination with ß-cell autoantibody determination, will improve the diagnosis of type 1 diabetes.

Item ID: 984
Item Type: Article (Refereed Research - C1)
Keywords: Type 1 Diabetes, Autoantigen, GAD65,ELISPOT assay
Additional Information:

Copyright © 2002 by the American Diabetes Association

ISSN: 1935-5548
Date Deposited: 02 Nov 2006
FoR Codes: 11 MEDICAL AND HEALTH SCIENCES > 1107 Immunology > 110703 Autoimmunity @ 0%
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