INTEGRINS AND T CELL DEVELOPMENT AND FUNCTION

该提案的总体目标是确定以下角色： 整合素受体与T淋巴细胞的发育和功能 选择了体内和体外相结合的方法。在……里面 为了测试整合素受体功能在T细胞中的作用 随着转基因小鼠的发展，已经产生了一种显性的 整合素阴性形式。具体地说，这个实验室使用的是 连接有活性整合素β1胞浆结构域的构建 结合到tac抗原的胞外区，这会破坏 细胞外基质与血管内皮细胞之间的跨膜连接 整合素受体提供的细胞骨架。近侧LCK 启动子已经被选择来指导胸腺特异性表达该基因 在转基因小鼠中构建，导致了 这些小鼠中成熟的单个阳性胸腺细胞与 正常的生命周期。我们的假设是，这一赤字代表着缺乏 在转基因小鼠中的正选择，我们将测试 利用单等位基因表达T的转基因小鼠的假说 细胞受体(TCR)具有明确的抗原识别功能。取决于 遗传背景、条件可以建立在哪些成熟 胸腺细胞只有通过单等位基因的正选择才能产生 克隆人。由于已显示肯定选择需要P2LRA， 表达显性阴性形式接头的转基因小鼠 蛋白Shc将被产生，它应该会阻止整合素信号转导 P21 ras。在体外，分析整合素信号的分子方法 T细胞共刺激功能的途径将被完成。vbl.使用 最近开发的记者构建，对FAK的作用 TCR整合素的激活在共刺激中所选择的 ECM底物，以及该信号在慢性疾病中可能发挥的作用 炎症性疾病。

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.