BIOCHEMICAL CHARACTERIZATION OF TH CELL ACTIVATION

TH 细胞激活的生化特征

• 批准号: 3183150
• 负责人: DAVID J MCKEAN
• 金额: $ 18.43万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-04-01 至 1991-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3183150
• 关键词: T lymphocyte; calcium; chromatography; crosslink; gene expression; genetic transcription; genetic translation; hydrolysis; immunologic techniques; immunoregulation; interleukin 1; interleukin 2; ionomycin; ionophores; laboratory mouse; leukocyte activation /transformation; lymphoma; membrane activity; messenger RNA; mitogens; molecular cloning; monoclonal antibody; phorbols; phosphatidylinositols; phospholipase C; phosphorylation; protein kinase C; radioassay; receptor; second messengers; tumor promoters

A comprehensive analysis of the role of IL1 on the antigen specific activation of class II-restricted cloned murine T lymphocytes will be done using two complementary T cell activation model systems. One system will utilize recombinat IL1 to enchance the activation of cloned, soluble antigen or alloantigen reactive T lympocytes stimulated in the presence of cloned B lymphoma accessory cells have been shown to activate T cell clones in antigen specific but IL1 independent fashion. The second module system will utilize combinations of recombinant IL1 and specific pharmacological modifiers of Ca++ (ionomycin) and protein kinase C (TPA, synthetic diacylglycerol) second messenger pathways to activate antigen and alloantigen reactive T cell clones. We have demonstrated that these soluble mediators effectively synergize with ionomycin in this system to elicit T cell activation responses. T cell clones which are either IL1 responsive or IL1 unresponsive in both assay systems have been identified and will be used as positive and negative experimental models. With these model systems we will quantitatively evaluate the effects of rIL1 and the parmacological modifiers on: A. The expression on the release of IL2 and of cell surface receptors that are directly involved in T cell activation, B. Activation of the phospholipase C-mediated hydrolysis of phosphoinositides, C. Identification of proteins which are phosphorylated as a result of the various initiators of T cell Activation, and D. Characterizartion by Northern blot and nuclear run off analyses of mRNA transcription which results from the various initiators of T cell activation. These analyses should significantly add to our understanding of the role of IL1 in regulating an antigen specific immune response. In addition, valuable information concerning the specific intracellular effects that may be related to cell transformation will be obtained using the tumor promoting phorbol esters (e.g. TPA).

对IL1在抗原特异性方面的作用的全面分析 将进行II类限制的克隆鼠T淋巴细胞的激活 使用两个互补的T细胞激活模型系统。 一个系统将 利用重组IL1来掩盖克隆的可溶性激活 在存在下刺激的抗原或同种反应性T淋巴细胞 克隆的B淋巴瘤辅助细胞已被证明激活T细胞克隆 以抗原特定但IL1独立的方式。 第二个模块系统 将利用重组IL1和特定药理的组合 Ca ++（离子霉素）和蛋白激酶C（TPA，合成的修饰剂）的修饰剂 二酰基甘油）第二信使途径激活抗原和 同种反应性T细胞克隆。 我们已经证明了这些 可溶性介体在该系统中有效地与离子霉素协同 引起T细胞激活反应。 T细胞克隆是IL1 在两个测定系统中响应敏感或IL1无反应性 并将用作正面和负面的实验模型。 与这些 模型系统我们将定量评估RIL1和 瘫痪者修饰符：A。IL2和IL2释放的表达 直接参与T细胞活化的细胞表面受体的 B.磷脂酶C介导的水解的激活 磷酸肌醇，C。鉴定磷酸化的蛋白质 由于T细胞激活的各种发起人和D。 Northern印迹的特征和核能流失mRNA的分析 转录来自T细胞的各个发起者 激活。 这些分析应大大增加我们的理解 IL1在调节抗原特异性免疫反应中的作用。 在 此外，有关特定细胞内的有价值的信息 可能与细胞转换有关的效果将使用 促进佛波酯的肿瘤（例如TPA）。