Natural Killer Cell Immune Synapse

描述(由申请人提供)：这些项目的总体目标 研究的目的是深入了解发生在单个 自然杀伤(NK)细胞与细胞相互作用时的细胞水平 敏感和不敏感的靶细胞。细胞间的接触区域， 称为免疫突触(IS)，经历时间和空间的变化 被认为对优化细胞相互作用至关重要 并控制结果。大多数关于信息系统的研究都是在Helper的帮助下进行的 T细胞与抗原提呈细胞(APC)相互作用。形成的组织 辅助T细胞/APC(这)演示了一个严格控制的过程，包括 信号分子顺序地招募到内部隔区 细胞间的接触地点。我们建议的研究的长期目标是 为了验证在细胞溶解的NK细胞相互作用过程中形成的假设 与目标单元格(NKIS)显示了与此的重要区别 在克隆性T细胞增殖启动过程中发生。我们将在未来 本方案利用MHC-I类分子调控的NK细胞杀伤模型 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 一种溶细胞性NKIS和一种非溶细胞性NKIS。我们假设一个截然不同的 信号转导分子的子集将被招募到细胞溶解 NKIS。具体地说，我们希望确定不同的信号招募 介导颗粒胞吐的分子，而酶和接头分子 参与细胞增殖将从突触中消失或移位 来自活动的信令组件。我们还假设，抑制性NK细胞 具有MHC-I类分子特异性配体的受体调节 通过配体诱导突触区域运动的信号通路。最后， 我们假设肌动蛋白-细胞骨架的聚合是由 Wiskott-Aldrich综合征蛋白(WASP)不是 NK细胞介导的细胞毒效应事件。该提案有三个方面 具体目标：(1)表征MHC I类受监管的时间和 突触内信号转导分子的空间组织 NK细胞与靶细胞的非溶细胞和溶细胞相互作用。(2)至 NK受体及其受体的时空组织特征 NK细胞-靶细胞结合物接触区的HL A配体 溶细胞性和非溶细胞性组合。(3)时代性和时代性 溶细胞性和非溶细胞性NKIS的空间组织 WASP基因的不同突变。我们希望这些研究能提供新的 深入了解NK细胞如何与靶细胞相互作用以及MHC-I类细胞如何 分子调节这些过程。

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.