THE IMMUNOLOGICAL STUDY OF GULUTAMIC ACID DECARBOXYLASE SPECIFIC T LYMPHOCYTES IN THE ETIOLOGY OF JUBENILE DIABETES

谷氨酸脱羧酶特异性T淋巴细胞在青少年糖尿病病因中的免疫学研究

• 批准号: 08670906
• 负责人: KAWAMURA Tomoyuki
• 金额: $ 1.47万
• 依托单位: OSAKA CITY UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08670906/
• 关键词: type 1 diabetes; T lymphocyte; glutamic acid decarboxylase; cytokines; type 1 helper T cell; I型糖尿病; I型ヘルパーT細胞; IDDM; リンパ球; GAD; インスリン依存性糖尿病

Type I diabetes is thought to be an autoimmune disease mediated by T lymphocytes against pancreatic beta cells. Recently, glutamic acid decarboxylase (GAD) was identified as a major autoanitigen of antibody existing in Type 1 diabetes patients. In this study, we focused on the T lymphocytes against GAD in Japanese Type 1 diabetes.Tlymphocytes isolated from peripheral blood were cultured with insulin, rat insulinoma cell (RIN) homogenate, or GAD in 10% AB type human serum contained RPMI medium for 7 days. At 7days, ^3H-Thymidine uptake was measured. T lymphocyte response against GAD in Type 1 diabetes patients was significantly stronger than that in healthy controls. The significant response against insulin was not observed. The response against RIN showed higher tendency in the patients but not significant. The secretion of cytokines in the supernatant during the response against GAD were measured by ELISA.In results, the lymphocytes secreted gamma-interferon but interleukin 4.We tried to make the GAD specific T cell clones but failed at the present time, but succeeded to make the GAD specific T cell lines. The flow cytometry analysis revealed that these cell lines were composed of CD4^+ Tlymphocytes and CD8^+ Tlymphocytes.These results indicated that GAD specific CD4^+ and CD8^+ T lymphocytes exist in the peripheral blood of Type diabetes I patients. The cellular immunity against GAD may play a role in the pathogenesis of Type 1 diabetes.

I型糖尿病被认为是一种由T淋巴细胞介导的针对胰腺β细胞的自身免疫性疾病。谷氨酸脱羧酶（GAD）是1型糖尿病患者体内存在的一种主要的自身抗体。本研究以日本1型糖尿病患者外周血T淋巴细胞为研究对象，将其与胰岛素、大鼠胰岛素瘤细胞（RIN）匀浆或GAD在含10%AB型人血清的RPMI培养基中培养7天。在第7天，测量^3H-胸苷摄取。1型糖尿病患者T淋巴细胞对GAD的反应明显强于健康对照组。未观察到对胰岛素的显著反应。对RIN的反应在患者中显示出更高的趋势，但不显著。结果表明，淋巴细胞分泌γ-干扰素，但不分泌白细胞介素4（IL 4）。我们尝试了制备GAD特异性T细胞克隆，但目前未能成功，但成功地制备了GAD特异性T细胞系。流式细胞仪分析表明，这些细胞系均由CD 4 ^+ T淋巴细胞和CD 8 ^+ T淋巴细胞组成，表明I型糖尿病患者外周血中存在GAD特异性的CD 4 ^+和CD 8 ^+ T淋巴细胞。抗GAD的细胞免疫可能在1型糖尿病的发病中起一定作用。