CHARACTERISTICS OF T CELL RECEPTORS

此应用程序旨在研究T细胞的各种属性 感受器。我们将集中研究识别抗原的受体。 与主要组织相容性复合体(MHC)产物相关 (抗原/H-2)。我们的最终目标是识别和隔离 负责这种识别和定位的分子(S) 从基因上来说。最近的研究将极大地帮助我们进行这些研究 克隆T细胞制备技术的实验室研究进展 识别抗原/H-2产物的杂交瘤。这些杂交瘤生长 在正常组织培养基上连续快速培养。当被质问时 然而，在适当的抗原/H-2结合下，它们会分泌 白介素2。它们还与抗原冲击的提呈细胞结合 适当的H-2单倍型。我们将尝试将这些特征 抗原/H-2受体(S)免疫化学和生物化学。免疫化学 分析将包括对抗血清和B杂交瘤的研究 T细胞杂交瘤，特别强调那些阻断 抗原结合。生化分析将包括尝试分离 来自放射性标记细胞的抗原和/或H-2结合物质。在……里面 与胡德博士合作，我们将研究免疫球蛋白的表达 这些细胞中有类重链的mRNA。磷酰胆碱(PC)反应性 将制作杂交瘤来检查编码VH基因和mRNA的状态 PC结合重链。在与Riblet博士的合作研究中，我们将 制备可用于重、卡帕和拉姆达分析的杂交物 基因重排。体细胞遗传学将被用来确定 代表抗原/H-2的T细胞受体(S)的染色体编码。T细胞 杂交瘤将用抗原特异的亲本T细胞和 标记的染色体，以及保留或丢失抗原的亚系 专一性，专一性和标记的染色体的一致性 请注意。此外，在与弗兰克博士的合作下，抗原 已经存在的杂交瘤的非反应性变体将被分析 染色体丢失。

This application is designed to study various properties of T cell receptors. We will concentrate on those receptors which recognize antigen in association with major histocompatibility complex (MHC) products (antigen/H-2) in the mouse. Our ultimate goals are to identify and isolate the molecule(s) responsibile for this recognition and to map them genetically. We will be greatly helped in these studies by the recent development in our laboratory of techniques for preparing cloned ?T cell hybridomas which recognize antigen/H-2 products. These hybridomas grow continuously and rapidly in normal tissue culture medium. When confronted with the appropriate antigen/H-2 combination, however, they secrete interleukin-2. They also bind to antigen-pulsed presenting cells of the appropriate H-2 haplotype. Attempts will be made to characterize the antigen/H-2 receptor(s) immunochemically and biochemically. Immunochemical analysis will include studies on antisera and B hybridomas raised against the T cell hybridomas, with particular emphasis on those which block antigen binding. Biochemical analysis will include attempts to isolate antigen and/or H-2 binding material from radiolabelled cells. In collaboration with Dr. Hood we will investigate the expression of Ig heavy-like mRNA in these cells. Phosphoryl choline (PC)-reactive hybridomas will be made to check the status of VH genes and mRNA coding for PC-binding heavy chains. In collaborative studies with Dr. Riblet we will prepare hybrids which can be used for analysis of heavy, kappa and lambda gene rearrangements. Somatic cell genetics will be used to establish which chromosomes code for the T cell receptor(s) for antigen/H-2. T cell hybridomas will be prepared using antigen specific parental T cells with marked chromosomes, and in sublines which retain or have lost antigen specificity the coincidence of specificity and the marked chromosomes will be noted. In addition, in collaboration with Dr. Francke, antigen non-reactive variants of already existing hybridomas will be analyzed for chromosome loss.