THE GADS ADAPTOR PROTEIN IN T CELL-MEDIATED PREVENTION OF VIRAL PATHOGENESIS

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。T细胞介导的抗病毒免疫依赖于TCR和CD28信号的成功转导。TCR复合体包含抗原识别和信号基序。CD28共受体传递关键的第二个信号，如果没有这个信号，细胞将变得无能并经历凋亡。第二个信号确保TCR的抗原刺激在适当的免疫环境中发生。这个研究项目的目标是研究CD28介导的信号，这些信号是T细胞激活所必需的。来自我们实验室和其他实验室的证据表明，CD28和GADS接头蛋白通过Akt丝氨酸/苏氨酸激酶调节生存和增殖。我们假设Gads、CD28和Akt在一个共同的信号通路中相连，这是正常T细胞发育和激活所必需的。为了验证这一假设，我们将重点关注三个具体目标。具体目的1描述了利用GADS缺陷小鼠来检测CD28介导的Akt磷酸化。特殊目的2将测试一个假设，即一个四元信号复合体连接CD28、Gads、PI3-K和c-Cbl，可能导致Akt磷酸化增强。描述了检验这种假定的突变复合体的生物化学实验。最后，在特定目的3中描述的实验应用遗传技术来检查CD28和GADS之间的联系。将分析Gads缺陷、CD28缺陷和Gads/CD28双缺陷小鼠的T细胞发育。总之，本提案中详细介绍的实验将检查连接CD28、Gads、PI3-K和c-Cb1的信号通路。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. T cell-mediated immunity against viral pathogenesis is dependent on the successful transduction of signals from the TCR and CD28 into the cell. The TCR complex contains the antigen recognition and signaling motifs. The CD28 co-receptor delivers a critical second signal, without which the cell would become anergic and undergo apoptosis. The second signal ensures that the antigenic stimulation of the TCR occurs in the appropriate immune context. The goal of this research project is to examine the CD28-mediated signals that are required for T cell activation. Evidence from our laboratory and others suggest that CD28 and the Gads adaptor protein regulate survival and proliferation through the Akt serine/threonine kinase. We hypothesize that Gads, CD28, and Akt are linked in a common signaling pathway that is required for normal T cell development and activation. To test this hypothesis, we will focus on threee specific aims. Specific aim 1 describes the use of Gads-deficient mice to examine CD28-mediated Akt phosphorylation. Specific aim 2 will test the hypothesis that a quaternary signaling complex links CD28, Gads, PI3-K, and c-Cbl that may lead to enhanced Akt phosphorylation. Biochemical experiments examing this putative mutlimeric complex are described. Finally, experiments described in specific aim 3 apply genetic techniques to examine the link between CD28 and Gads. T cell development in Gads-deficient, CD28-deficient, and Gads/CD28 double deficient mice will be analyzed. In summary, the experiments detailed in this proposal will examine the signaling pathway that links CD28, Gads, PI3-K, and c-Cbl.