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

青少年胰岛素依赖型糖尿病易感性 （IDDM）与两个HLA DQ等位基因-HLA DQ 8和HLA DQ 2连锁。因为 分离DQ限制性人T细胞克隆的巨大困难， 关于DQ限制性T细胞应答的特异性的信息， 胰岛细胞抗原不存在。 易感HLA DQ等位基因在DQB位置57处缺乏天冬氨酸。（DQ8） 在结合许多肽结合方面表现出显著的差异， 与DQ 9相比，DQ 9是DQ β 57 Asp+，其他方面与DQ 8相同。 T/H2 CD 4 + T/H1 T细胞导致胰岛β细胞破坏。胰岛抗原 特异性T/h2细胞是保护性的。DQ相关的一种机制 易感性是易感等位基因结合并呈现不同 诱导主要T/H1应答的肽的子集。耐 结合其他胰岛抗原肽的等位基因，其诱导T/h2 应答 拟议的实验将测试是否敏感和耐药 等位基因结合并呈递不同的肽子集。为此将 通过产生表达相关HLA DQ的转基因小鼠， 等位基因，以及人CD 4，在存在和不存在鼠CD 4的情况下。 HLADQ8.huCD4转基因小鼠与NOD回交5次， 患上I型糖尿病转基因小鼠和非转基因同窝仔 将监测对照的自发发展， 胰岛细胞抗原抗体和HLA DQ限制性T细胞应答 GA65和前胰岛素原。未处理的转基因小鼠和转基因小鼠 用重组人GD 65和前胰岛素原免疫的小鼠将用于 评估由DQ 8、DQ 7和DQ 6呈递给T细胞的肽。这将是 通过使用这些小鼠的淋巴结和脾脏T细胞进行小鼠 T细胞杂交瘤，DW 5147。将分析所得杂交瘤的 HLA DQ限制和肽特异性。 最后，由HLA DQ 8呈递的那些肽表位将用于 检测糖尿病前期和近期发病的HLA DQ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.