HIV-1 Suppression by MHC class I restricted CD8 T Cells

描述(申请人提供)：虽然动物模型研究指出CD8 T细胞反应在控制HIV-1复制中的重要性，但实现这一控制的这些细胞的具体质量尚不清楚。许多研究试图将HIV-1特异性CD8 T细胞的不同参数与疾病进展的临床标志相关联；然而，获得的数据并不一致。此外，对于已经发现与血浆病毒载量相关的研究，尚不清楚所观察到的CD8T细胞参数是否只是病毒复制低的结果，而不是控制这种复制的原因。这个问题源于这样一个事实，即大多数检测方法没有测量CD8 T细胞在控制HIV-1中的实际功能，而是测量了这种控制的潜在替代标记物。我们建议的具体目标1将使用体外病毒检测来分析HIV-1CD8 T细胞克隆在杀死和抑制HIV-1复制方面的效率。每个克隆都将根据T细胞受体基因(TCRB)的序列进行鉴定。利用多色流式细胞术对HIV-1特异性CD8 T细胞进行分类，根据其表达4种不同功能参数(干扰素-β、IL-2、肿瘤坏死因子-α和CD107)的能力进行分类。然后，我们将确定哪些特定的HIV-1 CD8 T细胞群富含了在体外有效控制病毒的克隆。我们的发现将对HIV免疫发病机制和疫苗设计具有重要意义，因为我们提供了HIV-1特异性CD8 T细胞表型，从而导致有效地控制HIV-1。

DESCRIPTION (provided by applicant): While animal model studies point to the importance of CD8 T cell responses in controlling HIV-1 replication, the specific quality of these cells achieving this control are unknown. Numerous studies have attempted to correlate different parameters of HIV-1 specific CD8 T cells with clinical markers of disease progression; however, inconsistent data has been attained. Also, for the studies that have seen correlations with plasma viral load, it is unclear whether the observed CD8 T cells parameter are just the result of low viral replication rather than a causative factor in controlling this replication. The problem stems from the fact that most assays do not measure the actual function of CD8 T cells in controlling HIV-1 but instead measure potential surrogate markers of this control. Specific Aim 1 of our proposal will analyze the efficiency of HIV-1 CD8 T cell clones in killing and suppressing HIV-1 replication using in vitro viral assays. Every clone will be identified on the basis T cell receptor ¿ gene (TCRB) sequencing. Using polychromatic flow cytometry in specific aim 2, we will sort HIV-1 specific CD8 T cells according to their capacity to express combinations of 4 different functional parameters (IFN-?, IL-2, TNF-a, and CD107). We will then determine which of the specific populations of HIV-1 CD8 T cells is enriched with clones that efficiently control virus in vitro. Our findings will have important implications fro HIV immunopathogenesis and vaccine design by supplying an HIV-1 specific CD8 T cell phenotype that results in efficient HIV-1 control.