BIOLOGY OF THE E-RECEPTOR ASSOCIATED ANTIGEN

E-受体相关抗原的生物学

• 批准号: 3130637
• 负责人: Malek Kamoun
• 金额: $ 13.56万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-07-01 至 1987-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3130637
• 关键词: T lymphocyte; cell bank /registry; cellular immunity; chromatography; clone cells; enzyme linked immunosorbent assay; flow cytometry; gel electrophoresis; gene expression; genetic library; human tissue; humoral immunity; immune adherence reaction; immunochemistry; interleukin 2; leukocyte activation /transformation; lymphokines; molecular cloning; monoclonal antibody; radiotracer; regulatory gene

Human T cells display a specific receptor (E) to sheep erythrocytes, causing the formation of spontaneous rosettes. Accumulating data suggest that the E-receptor and a p50 cell surface protein associated with it plays a regulatory role of multiple T cell functions that include the release of lymphokine(s) necessary for lymphocyte growth, cytotoxic T cell function and natural killer cell lysis. The long-term objectives of this proposal aim toward increasing our understanding of the biology of the p50 system. In particular we will test the hypothesis in which we propose that the E-receptor and p50 act as a modulator of both cellular and humoral immune reactions. Defective T cell E-rosette formation is observed in association with several diseases including cancer, autoimmune diseases and viral infections. This phenomenon appears to be mediated by serum factors that affect E-rosette formation and T cell function in a similar way than anti-p50 monoclonal antibody. A special emphasis will be given to experiments designed to analyze the mechanism by which E-receptor and p50 regulate T cell activation and the release of lymphokine(s) such as interleukin 2(IL-2). The effect of anti-p50 mAb will be analyzed using normal purified T cells, T cell subsets and various T cell clones. These experiments will include: (a) Kinetics of biosynthesis, maturation and release of p50 molecules of normal and mitogen-induced T cells using pulsechase labeling experiments and SDS gel electrophoresis of immunoprecipitated radio-labeled p50 proteins. (b) Cytofluometric analysis of the cell cycle of mitogen induced T cells, in particular the effect of anti-p50 on the G0-G1 transition will be examined. (c) Special emphasis will be given to examine the role of p50 in regulating IL-2 pathway: expression of IL-2 receptor (Tac antigen), release of IL-2 and more importantly the expression of IL-2 mRNA using cloned IL-2 cDNA probes. (d) In addition the role of p50 in regulating the release of lymphokine necessary for B cell growth, and immunoglobulin secretion will be examined using a B cell proliferation assay and an immunoglobulin ELISA assay respectively. (e) Finally, considerable effort will be made to clone p50 cDNA probes using synthetic oligonucleotide probes prepared on the basis of amino acid sequence analysis of purified p50 polypeptide. p50 cDNA probes will then be used to study the structure, function and expression of p50 gene products and its role in regulating T cell activation and lymphokine(s) productions.

人T细胞显示绵羊红细胞的特异性受体（E）， 从而导致自发性勃起。 不断积累的数据表明 E受体和与之相关的p50细胞表面蛋白 多种T细胞功能的调节作用，包括释放 淋巴细胞生长所必需的淋巴因子，细胞毒性T细胞功能 和自然杀伤细胞裂解。 本提案的长期目标 旨在增加我们对p50系统生物学的理解。 特别是，我们将测试我们提出的假设， E受体和p50作为细胞和体液免疫的调节剂 反应. 观察到T细胞E玫瑰花结形成缺陷 患有多种疾病，包括癌症、自身免疫性疾病和病毒性疾病， 感染. 这种现象似乎是由血清因子介导的， 影响E玫瑰花结形成和T细胞功能的方式与 抗p50单克隆抗体。 将特别强调 实验旨在分析E受体和p50 调节T细胞活化和淋巴因子的释放， 白细胞介素2（IL-2）。 抗p50 mAb的作用将使用 正常纯化的T细胞、T细胞亚群和各种T细胞克隆。 这些 实验将包括：（a）生物合成，成熟和 正常和丝裂原诱导的T细胞的p50分子的释放， 脉冲相位标记实验和SDS凝胶电泳 免疫沉淀放射性标记的p50蛋白。 (b)细胞荧光分析 有丝分裂原诱导的T细胞的细胞周期，特别是 将检查抗p50对G 0-G1转变的影响。 (c)特别强调 研究p50在调节IL-2通路中的作用： IL-2受体（Tac抗原）的表达，IL-2的释放等 重要的是使用克隆的IL-2cDNA探针的IL-2 mRNA表达。 （d）其他事项 此外，p50在调节淋巴因子释放中的作用 检查B细胞生长所必需的免疫球蛋白分泌 使用B细胞增殖测定和免疫球蛋白ELISA测定 分别 (e)最后，将作出相当大的努力克隆p50 cDNA探针使用基于以下制备的合成寡核苷酸探针： 纯化的p50多肽的氨基酸序列分析。 p50 cDNA探针 研究p50的结构、功能和表达 基因产物及其在调节T细胞活化和 淋巴因子的产生。