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

描述（由申请方提供）：虽然动物模型研究指出了CD 8 T细胞应答在控制HIV-1复制中的重要性，但实现这种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CD 8 T细胞的不同参数与疾病进展的临床标志物相关联;然而，已经获得了不一致的数据。此外，对于已经观察到与血浆病毒载量相关性的研究，尚不清楚观察到的CD 8 T细胞参数是否只是低病毒复制的结果，而不是控制这种复制的致病因素。这个问题源于这样一个事实，即大多数检测并不测量CD 8 T细胞在控制HIV-1中的实际功能，而是测量这种控制的潜在替代标志物。我们提案的具体目标1将使用体外病毒测定法分析HIV-1 CD 8 T细胞克隆在杀死和抑制HIV-1复制方面的效率。将根据T细胞受体基因（TCRB）测序鉴定每个克隆。在具体目标2中使用多色流式细胞术，我们将根据HIV-1特异性CD 8 T细胞表达4种不同功能参数（IFN-γ，IL-2、TNF-α和CD 107）。然后，我们将确定哪些特定的HIV-1 CD 8 T细胞群富含体外有效控制病毒的克隆。我们的研究结果将对HIV免疫发病机制和疫苗设计产生重要影响，通过提供HIV-1特异性CD 8 T细胞表型，从而有效控制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.