INTEGRINS AND T CELL DEVELOPMENT AND FUNCTION

整合素与 T 细胞的发育和功能

• 批准号: 2650060
• 负责人: EUGENE E. MARCANTONIO
• 金额: $ 20.04万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-30 至 1998-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2650060
• 关键词: T cell receptor; T lymphocyte; animal breeding; biological signal transduction; cytokine receptors; differentiation antigens; extracellular matrix; flow cytometry; genetically modified animals; immunogenetics; integrins; laboratory mouse; leukocyte activation /transformation; phenotype; receptor expression

The broad goals of this proposal are to determine the roles for integrin receptors in the development and function of T-lymphocytes. A combination of in vivo and in vitro approaches have been chosen. In order to test the role of integrin receptor function in T cell development, transgenic mice have been produced with a dominant negative form of integrin. Specifically, this laboratory is using a construct with the active integrin beta 1 cytoplasmic domain connected to the extracellular domain of the tac antigen, which breaks the transmembrane connection between extracellular matrix and the cytoskeleton provided by integrin receptors. The proximal lck promoter has been chosen to direct thymic specific expression of this construct in transgenic mice, leading to a reduction in the number of mature, single positive thymocytes in these mice as compared to normal liftermates. Our hypothesis is that this deficit represent a lack of positive selection in the transgenic mice, and we will test that hypothesis using transgenic mice with monoallelic expression of a T cell receptor (TCR) with a defined antigen recognition. Depending on the genetic background, conditions can be established in which mature thymocytes are produced only by positive selection of the monoallelic clones. Since positive selection has been shown to require p2lras, transgenic mice expressing a dominant negative form of the adaptor protein Shc will be produced, which should block integrin signaling to p21 ras. In vitro, molecular approaches to dissect integrin signaling pathways in co-stimulatory function of T cells will be done. Using recently developed reporter constructs, the role of versus FAK activation in of the TCR integrins have in co-stimulation on selected ECM substratum, and the role this signaling could play in chronic inflammatory diseases.

该提案的广泛目标是确定 整合素受体在T淋巴细胞的发育和功能中。 选择了体内和体外方法的组合。在 为了测试整联蛋白受体功能在T细胞中的作用 发育，转基因小鼠已经以优势产生 整联蛋白的负形式。具体来说，该实验室正在使用 用活跃的整合蛋白β1细胞质结构域连接的构造 到TAC抗原的细胞外结构域，这会破坏 细胞外基质与 整联蛋白受体提供的细胞骨架。近端LCK 已选择启动子来指导胸腺特异性表达 在转基因小鼠中构建，导致数量减少 这些小鼠的成熟，单个阳性胸腺细胞与 正常的LIFTERTS。我们的假设是这种赤字代表了缺乏 在转基因小鼠中的肯定选择，我们将测试 假设使用单相表达的转基因小鼠T 具有定义的抗原识别的细胞受体（TCR）。根据 遗传背景，可以建立成熟的条件 胸腺细胞仅通过单相阳性的阳性选择而产生 克隆。由于已显示阳性选择需要P2LRAS，因此 表达适配器的主要负面形式的转基因小鼠 将产生蛋白质SHC，这应阻止整联蛋白信号传导 P21 RAS。在体外，分子方法剖析整联蛋白信号传导 将完成T细胞共刺激功能的途径。使用 最近开发的记者构造，即FAK的作用 TCR整联蛋白的激活在选定的共刺激中具有 ECM基质以及该信号在慢性中的作用 炎症性疾病。