PATHOGENESIS AND PREVENTION OF TYPE I DIABETES IN THE NOD MOUSE AND MAN

NOD 小鼠和人 I 型糖尿病的发病机制和预防

• 批准号: 6270852
• 负责人: HUGH O MCDEVITT
• 金额: $ 13.14万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-02-15 至 1998-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6270852
• 关键词: MHC class II antigen; NOD mouse; autoantibody; disease /disorder etiology; genetically modified animals; glutamate dehydrogenase; insulin; insulin dependent diabetes mellitus; pancreatic islets

Susceptibility to juvenile onset insulin dependent diabetes mellitus (IDDM) is linked to two HLA DQ alleles--HLA DQ8 and HLA DQ2. Because of the great difficulty in isolating DQ restricted human T cell clones, information on the specificity of the DQ restricted T cell response to islet cell antigens is non-existent. Susceptible HLA DQ alleles lack aspartic acid in DQB position 57. (DQ8) exhibits striking differences in binding a number of peptide binding in comparison to DQ9, which is DQbeta57Asp+, and otherwise identical to DQ8. T/H2 CD4 + T/H1 T cells cause islet beta cell destruction. Islet antigen specific T/h2 cells are protective. One mechanism for DQ associated susceptibility is that susceptible alleles bind and present a distinct subset of peptides which induce a predominant T/H1 response. Resistant alleles binding other islet antigen peptides which induce a T/h2 responses. The proposed experiments will test whether susceptible and resistant alleles bind and present different subsets of peptides. This will be done by the production of transgenic mice expressing the relevant HLA DQ alleles, as well as human CD4, in the presence and absence of murine CD4. HLA DQ8.huCD4 transgenic mice at backcross 5 to NOD have been observed develop type I diabetes. Transgenic mice and non-transgenic littermate controls will be monitored for the spontaneous development of auto- antibodies to islet cell antigens and HLA DQ restricted T cell responses to GA65 and pre pro-insulin. Untreated transgenic mice and transgenic mice immunized with recombinant human GD65 and pre pro-insulin will be used to assess the peptides presented by DQ8, DQ7 and DQ6 to T cells. This will be done by using lymph node and spleen T cells from these mice to the murine T cell hybridoma, DW5147. The resulting hybridomas will be analyzed for HLA DQ restriction and peptide specificity. Finally, those peptide epitopes presented by HLA DQ8 will be used to detect HLA DQ restricted responses in pre-diabetic and recent onset diabetic patients.

对少年发作胰岛素依赖糖尿病的敏感性 （IDDM）链接到两个HLA DQ等位基因-HLA DQ8和HLA DQ2。由于 隔离DQ限制人类T细胞克隆的极大困难， 有关DQ限制T细胞响应对DQ的特异性的信息 胰岛细胞抗原不存在。 易感HLA DQ等位基因在DQB位置缺乏天冬氨酸57（DQ8） 在结合许多肽结合的结合中表现出明显的差异 与DQ9的比较，即DQBETA57ASP+，与DQ8相同。 T/H2 CD4 + T/H1 T细胞引起胰岛β细胞破坏。胰岛抗原 特定的T/H2细胞具有保护性。 DQ相关的一种机制 敏感性是易感等位基因结合并提出独特的 诱导主要T/H1反应的肽的子集。抵抗的 等位基因结合其他诱导T/H2的其他胰岛抗原肽 回答。 提出的实验将测试是否易感和抗性 等位基因结合并呈现不同的肽子集。这将完成 通过表达相关HLA DQ的转基因小鼠的产生 在存在和不存在鼠CD4的情况下，等位基因以及人CD4。 已经观察到Backcross 5至NOD的HLA DQ8.HUCD4转基因小鼠 发展I型糖尿病。转基因小鼠和非转基因窝窝 将监控控制措施，以自动开发自动发展 胰岛细胞抗原和HLA DQ限制T细胞反应的抗体 到GA65和前胰岛素前。未处理的转基因小鼠和转基因小鼠 用重组人GD65和前胰岛素免疫将用于 将DQ8，DQ7和DQ6提出的肽评估为T细胞。这将是 通过使用这些小鼠到鼠的淋巴结和脾脏T细胞完成 T细胞杂交瘤，DW5147。将分析所得的杂交瘤 HLA DQ限制和肽特异性。 最后，HLA DQ8提出的那些肽表位将用于 检测HLA DQ在糖尿病前和近期发作中的限制反应 糖尿病患者。