Natural Killer Cell Immune Synapse

自然杀伤细胞免疫突触

• 批准号: 6479000
• 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 2006-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6479000
• 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（由申请人提供）：这些的总体目标 研究的目的是深入了解单个事件中发生的分子事件 自然杀伤 (NK) 细胞以溶细胞作用相互作用时的细胞水平 敏感和非敏感靶细胞。细胞间接触面积， 称为免疫突触（IS），经历时间和空间的变化 被认为对于优化细胞相互作用至关重要 并控制结果。大多数 IS 研究都是在助手的帮助下进行的 T 细胞与抗原呈递细胞 (APC) 相互作用。的形成 辅助 T 细胞/APC IS (ThIS) 展示了一个严格调控的过程，涉及 信号分子顺序募集到内的分隔区域 细胞间接触部位。我们提出的研究的长期目标是 检验 IS 在细胞溶解性 NK 细胞相互作用过程中形成的假设 靶细胞 (NKIS) 与 ThIS 表现出重要差异 在克隆 T 细胞增殖起始期间发生。我们将在 目前的提案利用 MHC I 类调节的 NK 细胞细胞毒性模型 用于分析溶细胞性和非溶细胞性 NKIS。我们在我们的 初步结果表明这些事件与两者的形成有关 细胞溶解性 NKIS 和非细胞溶解性 NKIS。我们假设一个独特的 信号转导分子的子集将被招募到溶细胞 NKIS。具体来说，我们期望识别信号传导的差异招募 介导颗粒胞吐作用的分子，而酶和接头分子 参与细胞增殖的细胞将不在突触中或被移走 来自有源信号组件。我们还假设抑制性 NK 对 MHC I 类分子具有配体特异性的受体调节 配体诱导的突触区运动的信号通路。最后， 我们假设肌动蛋白-细胞骨架的聚合是由 威斯科特-奥尔德里奇综合征蛋白 (WASP) 不是 介导 NK 细胞的细胞毒性效应事件。该提案有三项 具体目标： (1) 表征 MHC I 类调节的时间和 突触区信号转导分子的空间组织 NK 细胞与靶细胞进行非溶细胞性和溶细胞性相互作用。 (2) 至 表征 NK 受体的时间和空间组织及其 NK 细胞-靶细胞缀合物接触区域的 HLA 配体 溶细胞性和非溶细胞性组合。 (3) 表征时间和 患者中细胞溶解性和非细胞溶解性 NKIS 的空间组织 WASP 基因的不同突变。我们期望这些研究能够提供新的 深入了解 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。