BIOLOGY OF THE E-RECEPTOR ASSOCIATED ANTIGEN

人类T细胞对绵羊红细胞显示一种特异的受体(E)， 导致自发的玫瑰花环的形成。越来越多的数据表明 E受体和与其相关的p50细胞表面蛋白所起的作用 多种T细胞功能的调节作用，包括释放 淋巴因子(S)对淋巴细胞生长、细胞毒T细胞功能的影响 和自然杀伤细胞裂解。这项建议的长期目标 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作为细胞免疫和体液免疫的调节剂 反应。相关观察到有缺陷的T细胞E花环的形成 包括癌症、自身免疫性疾病和病毒在内的几种疾病 感染。这一现象似乎是由血清因素介导的， 对E花环形成和T细胞功能的影响类似于 抗p50单抗。我们将特别强调 旨在分析E受体和p50的作用机制的实验 调节T细胞活化和淋巴因子释放(S)，如 白介素2(IL-2)。抗p50单抗的作用将用 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 人正常T细胞和丝裂原诱导的T细胞释放p50分子 脉冲酶标记实验和十二烷基硫酸钠凝胶电泳法 免疫共沉淀放射性标记的p50蛋白。(B)细胞荧光分析 对丝裂原诱导的T细胞的细胞周期的影响，尤其是对 将检测G0-G1转变上的抗p50。(C)特别强调 将被用来研究p50在调节IL-2途径中的作用： IL-2受体(Tac抗原)的表达、IL-2的释放等 重要的是用克隆的IL-2cDNA探针检测IL-2mRNA的表达。(D) 此外，p50在调节淋巴因子释放中的作用 B细胞生长所必需的，并将检查免疫球蛋白分泌 用B细胞增殖试验和免疫球蛋白酶联免疫吸附试验 分别进行了分析。(E)最后，将作出相当大的努力克隆p50 利用合成的寡核苷酸探针在下列基础上制备的cDNA探针 纯化的p50多肽的氨基酸序列分析。P50c DNA探针 将用于研究p50的结构、功能和表达 基因产物及其在调节T细胞活化和修复中的作用 淋巴因子(S)制作。

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.