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

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。T细胞介导的针对病毒发病机制的免疫依赖于TCR和CD28信号成功转导到细胞中。TCR复合物包含抗原识别和信号基序。CD28共受体传递一个关键的第二信号，没有这个信号细胞就会变得无能并发生凋亡。第二个信号确保TCR的抗原刺激发生在适当的免疫环境中。本研究项目的目标是检测T细胞活化所需的cd28介导的信号。我们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在正常T细胞发育和激活所需的共同信号通路中相互关联。为了验证这一假设，我们将关注三个具体目标。特异性目的1描述了使用gads缺陷小鼠来检测cd28介导的Akt磷酸化。特异性目的2将验证一个四元信号复合物连接CD28、Gads、PI3-K和c-Cbl的假设，这可能导致Akt磷酸化增强。描述了检验这种假定的多聚体复合体的生化实验。最后，在特定目标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，本提案中详细的实验将研究连接CD28、Gads、PI3-K和c-Cbl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.