Linking antigen-induced molecular membrane architecture to T cell responses

抗原(Ag)特异性T细胞和抗原提呈细胞(APC)之间的细胞相互作用可以触发 产生或流产的免疫反应。细胞的结果取决于T细胞的状态，他们的 以前接触过银，和APC。这一应用解决了中心假设，即分子 T细胞膜结构在影响抗原敏感性和细胞性方面起着决定性作用 结果。AIM 1将使用免疫电子显微镜、FRET和量子点闪烁来测试 NATve T细胞抗原刺激导致TCR微簇长期形成的假说 这有助于对召回AGS的反应。AIM 2将使用多维数字成像和生化 免疫学中识别特定抗原诱导的时空分子改变的分析 触发免疫反应改变的突触。这一目标将检验不成比例的假设 免疫突触中Fyn和Lck的激活可导致无反应，并将解决 免疫突触的功能意义。目标3将更直接地测试选择性的角色 Fyn或Lck对突触结构的激活。这一目标将使用新的无激酶T细胞和 可切换的突变失活酪氨酸激酶Fyn和Lck，可通过化学方法快速和 在T细胞中可逆地(在5分钟内)。 揭示T细胞感知抗原的机制以及这种识别如何决定 细胞反应是本计划所有5个项目的长期目标。有一个非常高的 与所有其他项目的协同效应。目标1与项目1协同工作，该项目将研究TCR集群 CDS细胞膜。AIM 2与项目3、4和5协同工作，这些项目将使用专门的成像技术 Sproutyl、TFII-I和CARD11各自的结构和功能研究中的工具及其与 免疫突触。AIM 3将与项目4协同，在该项目中，所有用于FYN的化学救援工具 而LCK将被开发出来。这些研究独一无二地结合了几个非常新颖的实验 这些方法很可能会大大提高对综合事件的理解，这些事件 对传染性病原体的免疫反应。这些知识可能会导致更有效的设计 疫苗，更好的免疫监测方案，以及新的免疫抑制药物。

Cellular interactions between antigen (Ag)-specific T cells and Ag-presenting cells (APCs) can trigger either productive or aborted immune responses. The cellular outcomes depend on the state of the T cells, their previous exposure to Ag, and the APCs. This application addresses the central hypothesis that the molecular architecture of the membrane of T cells plays a decisive role in influencing Ag-sensitivity and cellular outcomes. Aim 1 will use immuno-electron microscopy, FRET and Quantum Dot blinking to test the hypothesis that antigenic stimulation of naTve T cells results in long-term formation of TCR micro-clusters that facilitate responses to recall Ags. Aim 2 will use multi-dimensional digital imaging and biochemical analysis to identify specific antigen-induced spatial-temporal molecular alterations in Immunological Synapses that trigger altered immune responses. This Aim will test the hypothesis that disproportionate activation of Fyn and Lck in the Immunological Synapse can cause unresponsiveness and would address the functional significance of the Immunological Synapse. Aim 3 will test more directly the roles of selective activation of Fyn or Lck on the structure of the synapse. This Aim will use novel kinase-null T cellls and switchable mutated inactive tyrosine kinases Fyn and Lck that can be chemically rescued rapidly and reversibly (within 5 min) in T cells. Uncovering the mechanisms by which T cells can sense Antigen and how this recognition determines the cellular response is the long-term objective of all 5 projects in this Program. There is an extremely high synergy with all other projects. Aim 1 synergizes with Project 1 that will study TCR clustering in the membrane of CDS cells. Aim 2 synergizes with Projects 3, 4 and 5, which will use the specialized imaging tools in their own structure function studies of Sproutyl, TFII-I and CARD11 and their associations with the Immunological Synapse. Aim 3 would synergize with Project 4 where all the tools for chemical rescue of Fyn and Lck will be developed. These studies that combine uniquely the use of several highly novel experimental approaches are very likely to generate significant better understanding of the integrated events that regulate immune responses to infectious pathogens. This knowledge may lead to the design of more effective vaccines, better immune surveillance protocols, and new immunosupressive drugs.