Natural Killer Cell Immune Synapse

描述（由申请人提供）：总体目标 研究的目的是深入了解发生在单个细胞中的分子事件， 当自然杀伤（NK）细胞与细胞溶解性 易感和非易感靶细胞。细胞间接触面积， 称为免疫突触（IS），经历时间和空间变化， 被认为是至关重要的优化细胞相互作用 并控制结果大多数IS的研究都是在助手的帮助下进行的 与抗原呈递细胞（APC）相互作用的T细胞。形成 辅助性T细胞/APC IS（ThIS）展示了一个严格调节的过程， 将信号分子连续募集到 细胞间的接触点我们拟议研究的长期目标是 为了检验IS在溶细胞NK细胞相互作用期间形成的假设， 与靶细胞（NKIS）显示出与ThIS的重要差异， 在克隆性T细胞增殖的起始期间发展。我们会在 本发明利用MHC I类调节的NK细胞细胞毒性模型 用于分析溶细胞和非溶细胞NKIS。我们展示了我们的 初步结果表明，这些事件都与形成 溶细胞NKIS和非溶细胞NKIS。我们假设一种独特的 信号转导分子的子集将被募集到细胞溶解性细胞中。 NKIS。具体地说，我们希望确定不同的招聘信号 介导颗粒胞吐作用的分子，而酶和衔接分子 参与细胞增殖的细胞将从突触中消失或被取代 从活跃的信号组件。我们还假设抑制性NK细胞 对MHC I类分子具有配体特异性的受体调节 通过配体诱导的突触区域运动的信号通路。最后， 我们推测，肌动蛋白介导的细胞骨架聚合， Wiskott-Aldrich综合征蛋白（WASP）不是糖尿病的必需组分。 通过NK细胞介导细胞毒性效应物事件。该提案有三个 具体目的：（1）描述MHC I类调节的时间和 突触区信号转导分子的空间组织 NK细胞与靶细胞的非溶细胞和溶细胞相互作用。(2)到 表征NK受体的时间和空间组织及其 NK细胞-靶细胞偶联物接触区的HLA配体 溶细胞和非溶细胞组合。(3)为了描述时间和 患者中溶细胞和非溶细胞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 关于NK细胞如何与靶细胞相互作用以及MHC I类分子如何相互作用的见解 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.