Natural Killer Cell Immune Synapse

自然杀伤细胞免疫突触

• 批准号: 6865407
• 负责人: Bo Dupont
• 金额: $ 40.75万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-03-01 至 2008-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6865407
• 关键词: MHC class I antigen; Wiskott Aldrich syndrome; actins; antigen presenting cell; biological signal transduction; cell cell interaction; cellular immunity; clone cells; confocal scanning microscopy; cytotoxicity; digital imaging; enzyme activity; exocytosis; fluorescence resonance energy transfer; gene mutation; human tissue; laboratory mouse; leukocyte activation /transformation; mutant; natural killer cells; phosphatidylinositol 3 kinase; polymerization; protein kinase C; synapses

DESCRIPTION (provided by the applicant): The overall objective for these studies is to gain insight on the molecular events that occur at the single cell level when natural killer (NK) cells interact with cytolytically susceptible and non-susceptible target cells. The intercellular contact area, called the immune synapse (IS), undergoes temporal and spatial changes that are thought to be of critical importance for optimizing the cellular interactions and control the outcome. Most studies of the IS have been performed with helper T-cells interacting with antigen-presenting cells (APC). Formation of the helper T-cell/APC IS (ThIS) demonstrates a tightly regulated process involving sequential recruitment of signaling molecules to compartmentalized areas within the intercellular contact site. The long-term goal for our proposed studies is to test the hypothesis that the IS formed during cytolytic NK cell interactions with target cells (NKIS) display important differences from the ThIS that develops during initiation of clonal T cell proliferation. We will in the present proposal utilize the MHC class I regulated NK cell cytotoxicity model for analysis of the cytolytic and non-cytolytic NKIS. We demonstrate in our preliminary results that these events are associated with the formation of both a cytolytic NKIS and a non-cytolytic NKIS. We hypothesize that a distinct subset of signal transduction molecules will be recruited to the cytolytic NKIS. Specifically, we expect to identify differential recruitment of signaling molecules that mediate granule exocytosis while enzymes and adaptor molecules engaged in cell proliferation will be absent from the synapse or displaced away from active signaling components. We also hypothesize that the inhibitory NK receptors with ligand specificity for MHC class I molecules regulate the signaling pathways by ligand-induced movements in the synaptic region. Finally, we hypothesize that polymerization of actin-cytoskeleton mediated by the Wiskott-Aldrich Syndrome Protein (WASP) is not an essential component in mediating the cytotoxic effector events by NK cells. The proposal has three specific aims: (1) To characterize the MHC class I regulated temporal and spatial organization of signal transduction molecules in the synaptic region of NK cells in non-cytolytic and cytolytic interactions with target cells. (2) To characterize the temporal and spatial organization of NK receptors and their HLA ligands in the contact region of NK cell-target cell conjugates in cytolytic and non-cytolytic combinations. (3) To characterize the temporal and spatial organization of the cytolytic and non-cytolytic NKIS in patients with different mutations in the WASP gene. We expect these studies to provide new insight on how NK cells interact with target cells and how MHC class I molecules regulate these processes.

描述（申请人提供）：这些的总体目标 研究是为了洞悉单个发生的分子事件 当天然杀伤（NK）细胞与细胞溶剂相互作用时，细胞水平 易感和不敏感的靶细胞。细胞间接触区域， 称为免疫突触（IS），经历时间和空间变化 被认为对优化细胞相互作用至关重要 并控制结果。大多数IS研究都使用了助手进行 与抗原呈递细胞（APC）相互作用的T细胞。形成 助手T-cell/apc（这）表明了一个严格调节的过程，涉及 顺序募集信号分子以内部的区域化 细胞间接触站点。我们拟议的研究的长期目标是 测试在细胞溶解NK细胞相互作用期间形成的假设 与目标细胞（NKIS）一起显示与该目标细胞的重要差异 在启动克隆T细胞增殖期间的发展。我们将在 目前的建议使用MHC I类调控的NK细胞细胞毒性模型 用于分析细胞溶解和非溶解液的NKI。我们在我们的 这些事件与两者的形成相关的初步结果 细胞溶解的NKI和非溶解的NKI。我们假设这是一个独特的 信号转导分子的子集将募集到细胞溶解度 nkis。具体而言，我们希望确定信号的差异募集 介导颗粒胞吐作用的分子，而酶和衔接子分子 从突触中不参与细胞增殖或移位 来自主动信号组件。我们还假设抑制性NK MHC I类分子配体特异性的受体调节 配体引起的突触区域的运动信号通路。最后， 我们假设由 Wiskott-Aldrich综合征蛋白（WASP）不是必需的组成部分 介导NK细胞的细胞毒性效应事件。该提议有三个 具体目的：（1）表征MHC I类规范时间和 信号转导分子的空间组织在突触区域的突触区域 与靶细胞的非溶解和细胞溶解相互作用的NK细胞。 （2）至 表征NK受体及其的时间和空间组织 NK细胞目标细胞偶联的接触区域中的HLA配体在 细胞溶解和非溶解结合。 （3）表征时间和 患者患者的胞溶液和非溶解NKI的空间组织 黄蜂基因中的不同突变。我们希望这些研究能够提供新的 了解NK细胞如何与靶细胞相互作用以及MHC I类如何相互作用 分子调节这些过程。