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

青少年发病的胰岛素依赖型糖尿病的易感性 (IDDM)与两个HLADQ等位基因--HLADQ8和HLADQ2连锁。因为. 分离DQ的巨大困难限制了人类T细胞克隆， 关于DQ限制性T细胞应答特异性的信息 胰岛细胞抗原是不存在的。 易感的HLADQ等位基因在DQB 57位缺乏天冬氨酸。(DQ8) 在结合一些肽结合方面表现出显著的差异 与DQ9相比，它是DQbeta57Asp，在其他方面与DQ8相同。 T/H2CD4T/H1T细胞可导致胰岛β细胞破坏。胰岛抗原 特定的T/H2细胞具有保护性。DQ关联的一种机制 易感基因是指易感等位基因结合并呈现不同的 诱导主要的T/H1反应的多肽的子集。耐药 与其他胰岛抗原肽结合的等位基因可诱导T/H2 回应。 拟议的实验将测试是否敏感和耐药。 等位基因结合并呈现不同的多肽亚集。这件事会做到的 通过生产表达相关HLADQ的转基因小鼠 在存在和不存在小鼠的CD4的情况下，等位基因，以及人类的CD4。 观察到人类白细胞抗原DQ8.huCD4转基因小鼠在回交5至NOD时 患上I型糖尿病。转基因小鼠和非转基因小鼠 将监测自动控制的自发发展- 胰岛细胞抗原抗体和人类白细胞抗原DQ限制性T细胞反应 GA65和前胰岛素原。未经处理的转基因小鼠和转基因小鼠 重组人GD65和前胰岛素原免疫将用于 评估DQ8、DQ7和DQ6呈递给T细胞的多肽。这将是 通过使用这些小鼠的淋巴和脾T细胞给小鼠 T细胞杂交瘤，DW5147。由此产生的杂交瘤将被分析 人类白细胞抗原DQ限制和多肽特异性。 最后，这些由HLADQ8提出的多肽表位将被用于 检测糖尿病前期和新发患者的HLADQ限制性反应 糖尿病患者。

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.