T CELL RECEPTOR GENES IN AUTOIMMUNE DIABETES

T细胞在人类和动物发病机制中的重要性 胰岛素依赖型糖尿病（IDDM）已被充分 记录在案。 表达T细胞受体（TCR）的T细胞克隆型 与胰岛细胞自身抗原反应的基因 这样的假定 自身反应性TCR基因可能在基因组或细胞中检测到。 表达水平并因此反映在整个TCR库中， 在与相应的自身抗原反应的T细胞克隆型中， 和/或在疾病中发现浸润胰岛的那些 过程 NOD小鼠的IDDM样疾病提供并 一个很好的模型系统来评估这些可能性。 在 限制性片段长度多态性 在编码NOD小鼠的可变部分的基因中的限制性片段长度多态性（RFLP） TCR将用于遗传研究，以评估可能的 生殖系编码的TCR V基因对疾病的贡献 表现。 表型特征，特别是 胰腺癌细胞表达的克隆型TCR分子的性质 浸润性T细胞，将在RNA水平上使用V-PCR进行评估。 特异性分子探针，并在单细胞水平上使用 原位杂交和相关个体特异性抗体 TCR V基因亚家族。 如果像预期的那样， 在浸润细胞中鉴定TCR v基因使用 群体，反克隆型或变异型（V亚家族特异性） 特异于相关TCR分子的单克隆抗体将 在体内制备和给药，以评估其对 疾病过程。 这些模型研究将开始界定 小鼠糖尿病中假定的自身反应性T细胞克隆型 分子水平，并可能构成类似的基础 人类自身免疫性糖尿病的研究

The importance of T cells in the pathogenesis of human and animal insulin-dependent diabetes mellitus (IDDM) has been amply documented. T cell clonotypes expressing T cell receptor (TCR) genes reactive with islet cell autoantigens. Such putative autoreactive TCR genes might be detectable at the genomic or expression levels and thus reflected in the overall TCR repertoire, in the T cell clonotypes reactive with corresponding self-antigens, and/or those found infiltrating pancreatic islets in the disease process. The IDDM-like disease of the NOD mouse provides and excellent model system in which to assess these possibilities. In the proposed study, restriction fragment length polymorphisms (RFLPs) in the genes encoding the variable portion of the NOD mouse TCR will be utilized in genetic studies to assess the possible contribution of germline-encoded TCR V genes to disease manifestations. The phenotypic characteristics, and particularly the nature of the clonotypic TCR molecules expressed by pancreatic infiltrating T cells, will be assessed at the RNA level using V- specific molecular probes, and at the single-cell level using in situ hybridization and antibodies specific for relevant individual TCR V-gene subfamilies. If, as expected, a restricted pattern of TCR v. gene usage is identified in the infiltrating cell population, anti-clonotypic or variotypic (V subfamily-specific) monoclonal antibodies specific for the relevant TCR molecules will be prepared and administered in vivo to assess their effects on the disease process. These model studies will begin to define the role of putative autoreactive T cell clonotypes in murine diabetes at the molecular level, and might constitute the basis for similar studies in human autoimmune diabetes.