T CELL RECEPTOR GENES IN AUTOIMMUNE DIABETES

T细胞在人和动物发病机制中的重要作用 胰岛素依赖型糖尿病(IDDM)已经足够 有记录在案。表达T细胞受体的T细胞克隆型 与胰岛细胞自身抗原反应的基因。这样的推定 自身反应性TCR基因可能在基因组或 表达水平并且因此反映在整个TCR曲目中， 在与相应自身抗原反应的T细胞克隆型中， 和/或在疾病中发现的浸润性胰岛 进程。NOD小鼠的IDDM样疾病提供和 评估这些可能性的优秀模型系统。在……里面 拟议的研究--限制性片段长度多态 (RFLP)在编码NOD小鼠可变部分的基因中 TCR将被用于基因研究，以评估可能的 生殖系编码的TCRV基因在疾病中的作用 表现形式。表型特征，特别是 胰腺细胞表达的克隆型TCR分子的性质 将在RNA水平上使用V- 特定的分子探针，并在单细胞水平上使用In 原位杂交和相关个体特异性抗体 TCRV基因亚家族。如果如预期的那样，受限制的模式 TCRv.在浸润性细胞中确定了基因用途 群体，抗克隆型或变异型(V亚家族特有) 针对相关TCR分子的单抗将 准备并在体内给药，以评估它们对 疾病过程。这些模型研究将开始定义角色 糖尿病小鼠自身反应性T细胞克隆型的研究 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.