GENERATION AND EXPRESSION OF CYTOTOXIC T CELL FUNCTION

细胞毒性 T 细胞功能的产生和表达

• 批准号: 3125851
• 负责人: WILLIAM R CLARK
• 金额: $ 15.22万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-09-30 至 1990-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3125851
• 关键词: T lymphocyte; cell differentiation; cell membrane; cellular immunity; histocompatibility antigens; immunotoxicity; leukocyte activation /transformation; lipid structure; membrane activity; membrane structure; mixed lymphocyte reaction test; radiotracer; tissue /cell culture

The experiments in the present application are directed toward resolving: 1.) The mechanism of CTL-mediated lysis. The mechanism by which cytotoxic T lymphocytes kill specific target cells in still unknown. A study of lysis in lectin-mediated cytolysis (LDCC) led us to propose a new mechanism for CTL-mediated cytolysis, in which target cell death occurs as a result of distortion of target cell MHC proteins. Leakage at the interface between the transmembrane MHC proteins and surrounding lipids is postulated to lead to irreversible osmotic damage. Experiments are included in this proposal to test our hypothesis. Antibodies to MHC proteins, other transmembrane proteins, non-transmembrane proteins, and purely lipid antigens will be coupled to the surface of inert microspheres, microtiter wells, RBC or other nonlytic cell types. These will then be incubated with 51Cr-labeled target cells, and the rate of 51Cr release monitored. 2.) The relationship of lectin-mediated polyclonal activation of CTL function to LDCC and to antigen activation of CTL function. Certain plant lectins have been used both to activate (CTL function, and to mediate lysis in cases where MHC-restricted recognition is presumed not to occur. We have recently shown that target cell lysis in LDCC is in fact MHC-restricted, and we have obtained preliminary data indicating that lectin-mediated, polyclonal generation of CTL function is also controlled by MHC proteins involved in lectin presentation. We propose experiments to explore this point further, and to analyze its significance for CTL function generally. Using specific MHC antibodies, appropriate H-2 recombinant and congenic strains, and MHC positive and negative cell lines, we will probe the involvement of MHC proteins in activation of CTL function in primary and secondary reactions, in both syngeneic and allogeneic combination. We will also clone out individual CTL generated by lectin and compare their properties will CTL generated by specific antigen.

本申请中的实验旨在解决： 1)CTL介导的裂解机制。细胞毒作用的机制 T淋巴细胞对特定靶细胞的杀伤作用尚不清楚。关于……的研究 凝集素介导的细胞溶解(LDCC)中的裂解使我们提出了一种新的机制 对于CTL介导的细胞溶解，其结果是发生靶细胞死亡 靶细胞MHC蛋白的扭曲。界面处的泄漏 跨膜MHC蛋白和周围脂质之间的关系是假设的 导致不可逆转的渗透损伤。实验包括在这里面 用来检验我们假设的提案。抗MHC蛋白抗体，其他 跨膜蛋白、非跨膜蛋白和纯脂 抗原将偶联到惰性微球的表面，微滴度 Wells、RBC或其他非溶解细胞类型。然后，这些细胞将与 ~(51)Cr标记靶细胞，监测~(51)Cr释放率。2.) 凝集素介导的多克隆激活与CTL功能的关系 对LDCC和CTL功能的抗原活化有促进作用。某些植物凝集素 既被用来激活(CTL功能，也被用来中介裂解 推定不会发生MHC限制识别的情况。我们有 最近发现LDCC中的靶细胞裂解实际上是MHC限制的， 我们已经获得了初步的数据表明凝集素介导的， CTL功能的多克隆产生也受MHC蛋白的控制 参与凝集素的表达。我们提议用实验来探索这一点。 并对其对于CTL功能的意义进行了全面的分析。 利用特异性MHC抗体，适当重组和同源H-2 菌株，以及MHC阳性和阴性细胞系，我们将探索 MHC蛋白参与原发和非小细胞肺癌CTL功能的激活 二次反应，在同基因和同种异体组合中。我们会 也克隆出由凝集素产生的单个CTL并比较它们的 特性CTL会产生特定的抗原。