MHC CLASS II MOLECULES AND TYPE I IDDM

描述（改编自研究者摘要）：总体目的 这项研究应用的目的是了解MHC II类基因的作用， NOD小鼠对IDD的易感性和抗性。 前几 已发表的研究中，研究人员描述了MHC的影响 Ⅱ类基因多态性与NOD利用者IDD易感性的关系 携带来自IDD抗性株的Ab等位基因的转基因小鼠和突变的 Abg 7的形式。 PI的后续分析发现，这些研究 可能是有缺陷的，由于存在一个意想不到的影响， B细胞区室上的II类转基因表达。 在随后 实验中，PI已经开发出一种Abd转基因菌株， 以正常水平表达这些转基因而不影响B细胞 车厢 这种转基因导致IDD发病率的显著下降 在NOD中，与早期研究一致，并支持Abg 7在 IDD发病机制 当前应用程序的重点是评估角色 以及Abg 7介导IDD易感性的分子机制。 那里 有3个具体目标：1）分离T细胞克隆和T细胞杂交瘤 特异于GAD 65，如果可行，鼠前胰岛素原仅限于Ag 7 以及限制于Ag7.PD和Ag7.PD的类似的T细胞克隆和杂交瘤组。 Ad.在初步研究中，PI报告成功生产了 几个GAD反应性CD 4 + T细胞克隆。 这些T细胞将被用来 鉴定由这些II类MHC呈递的免疫显性肽表位 分子，并确定它们是否呈现不同的肽。 2)到 测量免疫显性GAD 65肽表位对每个 多聚甲醛固定的脾B细胞和单核细胞上的等位基因。 这些 研究将利用特定目标1中定义的肽来表征 这些肽与它们的相互作用的稳定性和亲和力 使用最初描述的肽结合程序 3）表征T细胞增殖反应，和 T细胞细胞因子产生模式，由每种免疫显性引起 肽结合到其各自的I-A等位基因。 这些研究旨在 评估这些不同的肽-II类等位基因复合物对 的T细胞应答的特征，并将该结果与 NOD的IDD发病机制。

DESCRIPTION (Adapted from Investigator's abstract): The overall objective of this research application is to understand the role of MHC class II genes in susceptibility and resistance to IDD in the NOD mouse. In previous published studies, the investigator has characterized the impact of MHC class II gene polymorphisms on IDD susceptibility in NOD utilizing transgenic mice carrying Ab alleles from IDD resistant strains and mutated forms of Abg7. Subsequent analyses by the PI have found that these studies may have been flawed due to the presence of an unexpected effect of MHC class II transgene expression on the B-cell compartment. In subsequent experiments, the PI has developed an Abd transgenic strain that appears to express these transgenes at normal levels without effecting the B-cell compartment. This transgene results in a dramatic decrease in IDD incidence in NOD, consistent with the early studies and supporting the role of Abg7 in IDD pathogenesis. The current application is focused on assessing the role and molecular mechanism by which Abg7 mediates IDD susceptibility. There are 3 specific aims: 1) To isolate T-cell clones and T-cell hybridomas specific for GAD65 and, if feasible, murine preproinsulin restricted to Ag7 and similar sets of T-cell clones and hybridomas restricted to Ag7.PD and Ad. In preliminary studies, the PI reports the successful production of several GAD reactive CD4+ T-cell clones. These T-cells would be utilized to identify the immunodominant peptide epitopes presented by these MHC class II molecules and to determine whether they present different peptides. 2) To measure the binding of the immunodominant GAD65 peptide epitopes for each allele on paraformaldehyde fixed spleen B cells and monocytes. These studies would utilize the peptides defined in Specific Aim 1 to characterize the stability and affinity of the interactions of these peptides with their respective alleles using the peptide binding procedure originally described by Rothbard et al. 3) To characterize the T-cell proliferative response, and pattern of T-cell cytokine production, elicited by each immunodominant peptide bound to its respective I-A allele. These studies are designed to assess the impact of these various peptide-class II allele complexes on the profile of the T-cell response elicited and to correlate this results with IDD pathogenesis in NOD.