SELECTIVE EXPRESSION OF LYMPHOKINES IN T LYMPHOCYTES

T 淋巴细胞中淋巴细胞因子的选择性表达

• 批准号: 2886049
• 负责人: MARIANNE T SWEETSER
• 金额: $ 8.75万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-04-01 至 2001-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2886049
• 关键词: gene expression; genetic regulatory element; helper T lymphocyte; interferon gamma; interleukin 4; laboratory mouse; methylation; protein biosynthesis; tissue /cell culture; transcription factor; transfection

Differential expression of lymphokines by CD4+ T cells plays a key role in proper modulation of the immune response. Consequently, aberrant control of lymphokine expression can have wide ranging impact on the progression of many infectious diseases such as tuberculosis and HIV as well as autoimmune diseases such as diabetes and multiple sclerosis. Th1 cells, which secrete IFN-gamma, IL-2 and lymphotoxin, direct delayed type hypersensitivity responses and are important in defense against intracellular organisms. Th2 cells, which produce IL-4, direct allergic or anti-inflammatory responses and protect against helminth infections. Regulation of IFN-gamma, IL-2 and IL-4 occurs primarily at the level of gene transcription, but the basis for the selective expression of these lymphokines by the various T cell subsets remains unclear. The overall objectives of this proposal are to define the molecular mechanisms by which IFN-gamma and IL-4 are differentially regulated by Th1 and Th2 CD4+ T cells and to develop an animal model in which the biological importance of developing the polarized population of T cells in response to infection and autoimmune disease can be assessed. Our recent studies have defined a key regulatory element in the IFN-gamma promoter which may contribute to the selective expression of IFN-gamma in Th1 cells and lack of expression in Th2 cells via interaction with factors which can mediate and repress expression and by methylation. The goal of this proposal is to use a two pronged approach to test this hypothesis. The first approach will utilize transient transfection of constructs containing this element into Th1 and Th2 cells to dissect the molecular pathway involved. The second concomitant approach will utilize in vivo promoter swapping experiments designed to determine the physiological importance of this element in the context of the endogenous gene and to address the role of methylation. These studies will build upon the candidate's previous training and allow her to gain significant new expertise in the generation and assessment of transgenic animals and in analysis of the complexity of the immune response in a whole animal context. These skills are essential for her development into an independent investigator. The sponsor's laboratory provides an ideal environment in which to pursue the proposed experiments since mouse embryo manipulation has been successfully utilized by this group and others at this institution and all the tools necessary to analyze the mice are available.

CD 4 + T细胞淋巴因子的差异表达起着关键作用 对免疫反应的适当调节。因此，异常 控制淋巴因子的表达可以对免疫系统产生广泛的影响。 许多传染病如结核病和艾滋病毒的发展 以及自身免疫性疾病如糖尿病和多发性硬化症。 Th 1细胞分泌IFN-γ、IL-2和α-光毒素， 迟发型超敏反应，在防御中很重要 对抗细胞内生物产生IL-4的Th 2细胞直接 过敏或抗炎反应，并防止蠕虫 感染. IFN-γ、IL-2和IL-4的调节主要发生在 基因转录的水平，但选择性的基础 这些淋巴因子由各种T细胞亚群表达仍然存在， 不清楚本提案的总体目标是确定 IFN-γ和IL-4差异表达的分子机制 受Th 1和Th 2 CD 4 + T细胞调节，并建立动物模型 其中发展两极分化的生物学重要性 T细胞群对感染和自身免疫性疾病的应答 可以评估。我们最近的研究已经确定了一个关键的调控机制， IFN-γ启动子中可能有助于选择性 IFN-γ在Th 1细胞中表达，而在Th 2细胞中缺乏表达 细胞通过与可以介导和抑制 表达和甲基化。该提案的目标是使用 两种方法来验证这个假设。第一种方法将 利用含有该元件的构建体的瞬时转染 进入Th 1和Th 2细胞，以剖析相关的分子途径。的 第二种伴随方法将利用体内启动子交换 旨在确定这一生理重要性的实验 在内源性基因的背景下，并解决的作用 甲基化。这些研究将建立在候选人以前的 培训，使她获得重要的新的专业知识， 转基因动物的产生和评估以及 免疫反应的复杂性。这些 技能是她发展成为一个独立的 调查员赞助商的实验室提供了理想的环境 在其中进行拟议的实验，因为小鼠胚胎 操纵已经被这个小组和其他人成功地利用， 在这个机构和所有必要的工具来分析老鼠是 available.