CHARACTERISTICS OF T CELL RECEPTORS

T 细胞受体的特征

• 批准号: 3128203
• 负责人: JOHN W KAPPLER
• 金额: $ 15.98万
• 依托单位: NATIONAL JEWISH HEALTH
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-05-01 至 1987-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3128203
• 关键词: B lymphocyte; T lymphocyte; cell transformation; gene expression; genetic mapping; hybridomas; immunochemistry; immunoglobulin genes; major histocompatibility complex; membrane activity; messenger RNA; monoclonal antibody; tissue /cell culture

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.

该应用旨在研究T细胞的各种特性 受体。 我们将专注于识别抗原的受体 与主要的组织相容性复合物（MHC）产品有关 （抗原/H-2）在小鼠中。 我们的最终目标是识别和隔离 该识别的分子责任并将其映射 遗传。 最近，我们将在这些研究中得到极大的帮助 在我们准备克隆细胞的技术实验室中开发 识别抗原/H-2产物的杂交瘤。 这些杂交瘤生长 在正常组织培养基中连续和快速。 当面对时 但是，使用适当的抗原/H-2组合，它们会分泌 白介素2。 它们还与抗原脉冲的呈现细胞结合 适当的H-2单倍型。 将尝试表征 抗原/H-2受体在免疫化学和生化上。 免疫化学 分析将包括对反对反对的抗血清和B杂交瘤的研究 T细胞杂交瘤，特别强调了阻止 抗原结合。 生化分析将包括试图隔离 来自放射性标记细胞的抗原和/或H-2结合材料。 在 与Hood博士的合作我们将研究IG的表达 这些细胞中的重型mRNA。 磷酸胆碱（PC） - 反应性 将使杂交瘤检查VH基因的状态和MRNA编码的状态 PC结合重链。 在与Riblet博士的合作研究中，我们将 准备可用于分析重型，Kappa和Lambda的杂种 基因重排。 体细胞遗传学将用于确定哪个 抗原/H-2的T细胞受体的染色体代码。 T细胞 杂交瘤将使用具有抗原特异性亲本T细胞制备 明显的染色体，在保留或失去抗原的subline中 特异性特异性和标记染色体的巧合将 被注意。 此外，与Francke博士合作，抗原 将分析已经存在的杂交瘤的非反应性变体 染色体损失。