Direct/Indirect NK Activation by HIV-1 Infected Cells

描述(由申请人提供)：本提案的总体目标是帮助理解HIV-1感染的靶细胞如何影响NK细胞的表型、激活状态和活性。HIV-1感染靶点可能通过多种方式影响NK细胞，包括HIV-1被膜与活化NK细胞上表达的趋化因子受体的相互作用，以及它们与HIV-1诱导的细胞因子表达在HIV-1疾病过程中的调节的关系。在第一个目标中，我们建议通过在体外将PBMC与感染HIV-1的自体CD4+初级T细胞孵育来分析NK细胞，并测试病毒诱导的NK激活状态、活性和增殖能力的调节。我们将评估CD107a暴露对靶细胞相互作用的NK活性的影响，并通过核糖核酸酶保护实验表征促炎症和凋亡相关基因的表达。在第二个目标中，我们将研究树突状细胞在NK识别和杀伤HIV-1感染的靶细胞中的辅助作用，以验证NK细胞在缺乏辅助细胞帮助的情况下裂解HIV感染靶细胞的能力受损的普遍假设。总体而言，这项研究将提高我们对艾滋病毒控制机制的理解，并可能确定新的干预目标。

DESCRIPTION (provided by applicant): The overall goal of this proposal is to contribute to the understanding of how HIV-1 infected target cells influence NK cell phenotype, activation state, and viability. HIV-1 infected targets may influence NK cells in several ways; including HIV-1 envelope interactions with chemokine receptors expressed on activated NK cells and their relation to HIV-1 induced modulation of cytokine expression during HIV-1 disease. In the first aim, we propose to analyze NKs by incubating PBMCs with autologous CD4+ primary T cells infected with HIV-1 in vitro and testing for virus-induced modulation of NK activation state, viability, and proliferative capacity. We will evaluate NK activation by CD107a exposure upon target cell interaction and characterize proinflammatory and apoptosis related gene expression by RNase protection assays. In the second aim, we will investigate the accessory role of dendritic cells in NK recognition and killing of HIV-1 infected target cells, in order to test the general hypothesis that NK cells are impaired in their ability to lyse HIV-infected targets in the absence of accessory cell help. Overall, this research will improve our understanding of mechanisms of control of HIV and potentially identify new targets for intervention.