SPECIFICITY/DISTRIBUTION/FUNCTION/GAMMA:DELTA T CELLS

这个项目的总体目标是确定其特殊性， 携带新发现T细胞的T细胞的分布和功能 包括与CD3相关的γ：Delta异二聚体的受体， 我们称之为TCR1。我们的工作假设是TCR1是 与传统的第I类MHC基因产物不同 在小鼠MHC(IB类)中的基因，它们的主要作用是 监测上皮细胞膜的完整性。为了测试这一点 假设，将进行以下实验： 将分离上皮内淋巴细胞并检测其表达情况 编码TCR基因的信使核糖核酸，并通过 用抗CD3或抗TCR抗体沉淀，并通过 细胞表面表型和受体表达的免疫荧光检测 在有试剂的地方。 分离的携带TCR1的T细胞将被抗原或 抗受体抗体，在IL-2中繁殖，并克隆和/或杂交 向TCR阴性的T淋巴瘤细胞提供稳定的TCR1来源 有T细胞。 新鲜分离的携带TCR1的T细胞或克隆株的功能 或者，将使用抗CD3抗体作为模拟配体来研究杂交细胞。这个 携带TCR1的静息和克隆T细胞的激活要求 将对淋巴因子的产生和反应进行研究 这些细胞的淋巴因子。 最后，这些细胞将被用来确定配体的特异性 L细胞刺激上皮细胞分离携带TCR1的T细胞 转导了多种MHC基因。如果这样的基因确实激活了TCR1 正常细胞、感染细胞和转化细胞中的表达 将对该组织进行检查，以确定是否存在差异表达 该基因可以解释TCR1介导的上皮监视。

The overall goal of this project is to determine the specificity, distribution, and function of Tcells bearing the newly described T cell receptor comprising the gamma:delta heterodimer associated with CD3, which we refer to as TCR1. Our working hypothesis is that TCR1 is specific for class I MHC gene products distinct from conventional class I genes in the murine MHC (class IB), and that their principal role is surveillance of the integrity of epithelial cell membranes. To test this hypothesis, the following experiments will be carried out: Intraepithelial lymphocytes will be isolated and tested for expression of mRNA encoding TCR genes, and for surface expression of TCR protein by precipitation with anti-CD3 or anti-TCR antibodies, and by immunofluorescence for cell surface phenotype and receptor expression where reagents are available. Isolated TCR1-bearing T cells will be stimulated with antigen or with anti-receptor antibody, propagated in IL-2, and cloned and/or hybridized to TCR-negative T lymphoma cells to provide a stable source of TCR1 bearing T cells. The function of freshly isolated TCR1 bearing T cells or of cloned lines or hybrids will be studied using anti-CD3 antibody as a mock ligand. The activation requirements of resting as well as cloned TCR1 bearing T cells will be studied, as will the production of lymphokines and the response to lymphokines of these cells. Finally, these cells will be used to determine the ligand specificity of TCR1 bearing T cells isolated from epithelia, by stimulation with L cells transfected with various MHC genes. If such a gene does activate a TCR1 bearing cell, its expression in normal, infected and transformed cells of that tissue will be examined to determine if differential expression of that gene could account for TCR1-mediated epithelial surveillance.