CELL BIOLOGY OF IMMUNE INTERACTIONS

免疫相互作用的细胞生物学

• 批准号: 6703661
• 负责人: Abraham Kupfer
• 金额: $ 20.63万
• 依托单位: NATIONAL JEWISH HEALTH
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-09-01 至 2004-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6703661
• 关键词: CD28 molecule; CD4 molecule; MHC class II antigen; T cell receptor; T lymphocyte; antigen presenting cell; binding proteins; biological signal transduction; cell cell interaction; cytoskeletal proteins; fluorescence resonance energy transfer; genetically modified animals; guinea pigs; laboratory mouse; laboratory rabbit; leukocyte activation /transformation; protein localization; protein structure function; protein tyrosine kinase

Cellular interactions between antigen (Ag)-specific T cells and antigen-presenting cells (APCs) can trigger either productive immune responses or aborted immune responses. These diverse biologic outcomes are determined by the complex integration of signals that are delivered upon the ligation of multiple receptors on the membrane of T cells with their counter receptors on the membrane of APCs. Uncovering the complex integrated cellular and molecular mechanisms, that determine the fate of the interacting cells, is the long term objective of these studies. Using novel 3-D digital imaging of T-APC conjugates it was recently shown that signaling and adhesion proteins are recruited to the cell contacts and form spatially segregated Supra-Molecular Activation Clusters (SMACs). This application is highly focused on the structure and function of the c-SMAC, which is the site of TCR engagement and activation. The central premise of this proposal is that TCR associated activation events are spatially and temporally orchestrated in the newly discovered c-SMAC. To test this hypothesis we will combine the use of T cell transgenic mice and multi-dimensional imaging and will define the 4-dimensional molecular translocations and association of four key activation receptors in the c-SMAC: TCR, CD45, CD4 and CD28 during T-APC interactions. These studies will be followed by derivative experiments to determine the mechanisms of association of each of these receptors with the c-SMAC and their physiological significance. The specific aims are: Aim 1. To determine the precise temporal and 3-dimensional molecular composition of c-SMACs in cell conjugates formed between T cells from TCR trangenic mice and B-APCs. Aim 2. To study the mechanisms that cause the transient colocalization of CD45 with Ag-bound TCR and Lck in early c-SMACs and to assess the functional significance of these translocations. Aim 3. To study the mechanisms that cause the transient early association of CD4 with the engaged TCR and Class II in c-SMACs, using TCR and CD4 tailless transgenic mice. Aim 4. To study the mechanisms responsible for the association of CD28 with the engaged TCR in the cSMAC and the role of c-SMAC-associated CD28 in T cell activation. These novel studies would provide the first 4-dimensional view of T-APC interactions and, in combination with genetic and functional studies, are very likely to generate significant better understanding of the integrated events that properly regulate immune responses. This new knowledge may be useful in future designs of better immune surveillance protocols, immunosupression drugs and new vaccines.

抗原(Ag)特异性T细胞与细胞间的相互作用 抗原提呈细胞(APC)可以触发任一生产性免疫 应答或中止的免疫应答。这些不同的生物结果 是由传递的信号的复杂集成决定的 在T细胞膜上的多个受体与 它们在APC膜上的反受体。揭开复杂的面纱 综合的细胞和分子机制，决定了 相互作用的细胞，是这些研究的长期目标。 利用新的T-APC偶联物的3-D数字成像，最近展示了 信号和黏附蛋白被招募到细胞接触 并形成空间分离的超分子激活簇(SMAC)。 本应用程序高度关注 C-SMAC，它是TCR参与和激活的部位。中环 该提议的前提是TCR关联的激活事件是 在新发现的c-SMAC中进行空间和时间上的协调。至 验证这一假设，我们将结合使用T细胞转基因小鼠和 多维成像将定义4维分子 四种关键激活受体在血管内皮细胞中的移位和关联 C-Smac：T-APC相互作用过程中的TCR、CD45、CD4和CD28。这些研究 之后将进行衍生实验，以确定 这些受体与c-Smac及其受体的关系 生理意义。具体目标是：目标1.确定 C-SMACs的精确时间和三维分子组成 TCR转基因小鼠的T细胞与B-APC之间形成的细胞偶联。 目的2.研究短暂性共定位的机制。 CD45与抗原结合的TCR和LCK在早期c-SMAC中的表达 这些易位的功能意义。目的3.研究 导致人类免疫缺陷病毒的短暂早期关联的机制 在c-SMAC中使用TCR和II类，使用TCR和CD4无尾转基因 老鼠。目的4.研究与人类健康相关的机制 CD28与cSMAC中参与的TCR和c-SMAC相关的作用 CD28在T细胞活化中的作用 这些新颖的研究将提供T-APC的第一个4维视图 相互作用，结合遗传和功能研究，是 极有可能显著地更好地理解集成 适当调节免疫反应的事件。这一新知识可能是 对未来设计更好的免疫监视协议很有用， 免疫抑制药物和新疫苗。