FAS ANTIGEN AND PERIPHERAL T CELL TOLERANCE

描述（改编自研究者摘要）：本研究的目标 应用是定义Fas信号通路在细胞凋亡中的作用。 正常小鼠外周耐受的维持和 系统性红斑狼疮（SLE）的lpr/lpr小鼠模型。 中央 这些研究的宗旨是，Fas在免疫中的主要功能位点， 调节是在外围，其中抗原经历的亚群 T细胞表达Fas，并对Fas刺激的凋亡敏感 自杀 这些研究的核心将是开发一种新的肽， 抗原特异性TCR转基因lpr/lpr小鼠模型，这将允许更多的 无反应性和凋亡缺失以及耐受性丧失的精确分析 与lpr表型的Fas缺陷有关。 双ab TCR转基因 在I-Ad的背景下与卵清蛋白（OVA）反应的抗体将被培育成 lpr/lpr小鼠以提供Fas缺陷型CD 4 + T细胞的可再生来源， 确定抗原特异性。 该系统将允许仔细研究Fas 和CD 28在体内和体外调节耐受机制。 在 与此同时，一组“人造”APC正在开发中， II类MHC的受控的组成型表达， CD 28/CTLA-4和Fas的反受体的协调表达 受体。 原位检测细胞凋亡的敏感技术 将与一种新的双标记原位杂交技术相结合 和流式细胞术分析以进行T细胞的单细胞分析， 在体外和体内操作的群体。

DESCRIPTION (Adapted from the Investigator's abstract): The goal of this application is to define the role of the Fas signaling pathway in the maintenance of peripheral tolerance in normal mice and in tolerance loss in the lpr/lpr murine model of systemic lupus erythematosus (SLE). The central tenet of these studies is that the principle site of Fas function in immune regulation is in the periphery, where a subpopulation of antigen-experienced T-cells express Fas and become susceptible to Fas-stimulated apoptotic suicide. Central to these studies will be the development of a new, peptide antigen-specific TCR transgenic lpr/lpr mouse model that will allow a more precise analysis of anergy and apoptotic deletion, and tolerance loss related to the Fas defect of the lpr phenotype. A double ab TCR transgene reactive with ovalbumin (OVA) in the context of I-Ad will be bred into lpr/lpr mice to provide a renewable source of Fas-deficient CD4+ T-cells of defined antigenic specificity. This system will allow careful study of Fas and CD28 regulation of tolerance mechanisms both in vivo and in vitro. In parallel, a panel of 'artificial' APCs is being developed that has controlled, constitutive expression of the class II MHC with or without coordinate expression of the counter-receptors for the CD28/CTLA-4 and Fas receptors. Sensitive techniques for in situ assay of apoptotic cell death will be coupled with a novel double-label in situ hybridization technique and flow cytometric analyses to perform single-cell analyses of T-cell populations manipulated in vitro and in vivo.