REGULATION LYMPHOCYTE PROLIFERATION AND DIFFERENTIATIO

调节淋巴细胞增殖和分化

• 批准号: 6497082
• 负责人: RANJAN SEN
• 金额: $ 27.51万
• 依托单位: BRANDEIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-01 至 2005-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6497082
• 关键词: T lymphocyte; apoptosis; biological signal transduction; cell differentiation; cell growth regulation; fluorescence microscopy; leukocyte activation /transformation; lymphocyte proliferation; nuclear factor kappa beta; pentoxifylline; phosphorylation; protein biosynthesis; tissue /cell culture; yeasts

DESCRIPTION (adapted from investigator's abstract): Attenuation of immune responses, caused by infection, age or genetic abnormality, leads to increased susceptibility to normally harmless pathogens. Conversely, aberrant activation of immune responses, such as in autoimmune diseases, is also severely detrimental to health. Thus, the balance between activation and inactivation of lymphocytes is essential for human well-being. Our long-term objective is to understand the molecular mechanisms that achieve this balance, and thereby design rational strategies to intervene in these processes. In this application we propose to investigate how NF-KB proteins participate in lymphocyte activation and inactivation. The Specific Aims are as follows: 1) to study for the molecular basis of down-regulation and cytoplasmic tethering of NF-kB proteins upon cessation of an activating signal. These studies will provide insights into the re-establishment of a resting slate after an immune response; 2) to study the mechanisms that govern the timing of activation-induced cell death (AICD) in T cells. We will examine the role of NF-KB in priming cells for AICD, and maintaining cell viability during the effector phase; 3) to characterize tissue-specific modifications of IkBa and the basis for differential association with Rel family members. These objectives will be accomplished by a combination of biochemical and genetic methods. Nuclear import and export of NF-KB will be assayed in yeast and mammalian cells using fluorescence microscopy; mutant yeast strains, and pharmacologic inhibitors will be used to study the role of IKB proteins in cytoplasmic retention and down-regulation of NF-kB. The role of NF-KB in AICD will be examined by uncoupling FasL induction from the induction of death. These studies will also seek a mechanism for the inhibition of AICD by the drug pentoxifylline. Lastly, phosphorylation of IK-Ba and its effects on c-Rel versus p65 association will be analyzed by mutagenesis of IkBa; putative kinases and phosphatase will be identified in vitro.

描述（改编自调查员的摘要）：免疫的衰减 由感染，年龄或遗传异常引起的反应导致增加 对通常无害病原体的敏感性。相反，异常激活 免疫反应（例如自身免疫性疾病）也很严重 对健康有害。因此，激活与失活之间的平衡 淋巴细胞对于人类的福祉至关重要。我们的长期目标是 了解达到这种平衡的分子机制，从而 设计理性策略以干预这些过程。 在此应用中，我们建议研究NF-KB蛋白如何参与 淋巴细胞激活和失活。具体目的如下：1） 研究下调和细胞质束缚的分子基础的研究 NF-KB蛋白停止激活信号。这些研究会 提供有关在免疫后重新建立休息板的见解 回复; 2）研究控制时间安排的机制 激活诱导的T细胞中的细胞死亡（AICD）。我们将研究 NF-kb在启动细胞中用于AICD，并在此期间保持细胞活力 效应阶段； 3）表征IKBA的组织特异性修饰 与REL家人差异关联的基础。 这些目标将通过生化和 遗传方法。 NF-KB的核进口和出口将在酵母中分析 使用荧光显微镜和哺乳动物细胞；突变的酵母菌菌株， 药理学抑制剂将用于研究IKB蛋白在 NF-KB的细胞质保留和下调。 NF-KB在AICD中的作用 将通过从诱导死亡诱导FASL诱导来检查。 这些研究还将寻求一种药物抑制AICD的机制 五氧化氨酸。最后，IK-BA的磷酸化及其对C-REL的影响 与p65的关联将通过IKBA的诱变进行分析；假定 激酶和磷酸酶将在体外鉴定。