Combining immunogenic peptides and Nef blockade to enhance CD8 T-cell-mediated clearance of HIV-infected cells

项目摘要 治疗期间HIV蛋白Nef的表达通过以下方式促进HIV在细胞中的持续 下调细胞表面主要组织相容性复合体I型(MHC-I)和抗原提呈 使病毒能够逃避免疫反应。因此，抑制Nef的表达将允许MHC-I HIV感染细胞表面HIV抗原的表达及其被HIV特异性CD8 T细胞的清除 除了MHC-I下调外，在开发有效的CD8细胞毒性T细胞(CTL)方面也存在挑战 对复制能力强的前病毒的反应。细胞内的艾滋病毒储存库变得由病毒主导 含有CTL逃逸突变的变体对免疫反应和缺陷的HIV前病毒具有抵抗力 可以产生病毒蛋白，作为CTL反应的诱饵。然而，最近的研究表明，多功能 CD8 T细胞的反应和/或针对结构上重要的(即，高度联网的)T细胞表位 而病毒蛋白的遗传保守区对艾滋病毒控制至关重要。此外，含有T细胞 通过将CD8 T细胞反应重定向至未突变的病毒表位，可消除细胞表位逃逸突变 通过采用一种创新的技术--体外艾滋病毒根除试验--使用参与者的细胞 在有效治疗方面，我们预计这项探索性研究将揭示Nef受体阻滞剂显著增强CD8 T细胞介导的含有诱导性前病毒的HIV感染细胞的消除。与Nef相结合 封锁，我们将通过扩增HIV特异性CD8 T细胞来增强这些HIV储存细胞的清除 诱导多功能/效应性CD8 T细胞反应的一组免疫原肽。这些多肽是 从六个HIV蛋白的拓扑重要和遗传保守区中选择 免疫信息学分析管道。由于这些多肽在它们的蛋白质中高度网络连接 起源，预计它们代表没有逃逸突变的T细胞表位。因此，在进行这项工作时， 研究我们将确定Nef阻滞剂和免疫原肽库的最佳组合以诱导CD8 T细胞介导的HIV感染细胞的清除。这项研究将加速开发一种新的艾滋病毒 将Nef阻断MHC-I修复和免疫疗法相结合的治疗策略 CTL反应，并为在该方法可以之前进展到体内模型提供证据基础 应用于临床环境。

Project Summary The expression of the HIV protein Nef during therapy contributes to the persistence of HIV in cells by downregulating cell-surface major histocompatibility complex type I (MHC-I) and antigen presentation which allows the virus to evade immune response. Therefore, inhibiting the expression of Nef would allow for MHC-I expression of HIV antigens on the surface of HIV-infected cells and their clearance by HIV-specific CD8 T cells. In addition to MHC-I downregulation, there are challenges in developing an effective CD8 cytotoxic T cell (CTL) response against replication-competent proviruses. The intracellular HIV reservoir becomes dominated by viral variants containing CTL escape mutations that are resistant to immune response and defective HIV proviruses can produce viral proteins that act as decoys for CTL response. However, recent studies show polyfunctional response of CD8 T cells and/or the targeting of T cell epitopes from structurally important (i.e., highly networked) and genetically-conserved regions of viral proteins are essential for HIV control. In addition, cells harboring T cell epitope escape mutations can be eliminated by redirecting CD8 T cell response to unmutated viral epitopes By employing an innovative technique-- an ex vivo HIV eradication assay --using cells from participants on effective therapy, we expect this exploratory study will reveal that Nef blockade significantly enhances CD8 T cell–mediated elimination of HIV-infected cells containing inducible proviruses. In combination with Nef blockade, we will augment the clearance of these HIV reservoir cells by expanding HIV-specific CD8 T cells with a pool of immunogenic peptides that induce polyfunctional/effector CD8 T cell response. These peptides are selected from topologically important and genetically-conserved regions of six HIV proteins by applying an immunoinformatics analysis pipeline. Due to the fact these peptides are highly networked within their protein of origin, they are expected to represent T cell epitopes which lack escape mutations. Therefore, in conducting this study we will determine the best combination of Nef blockers and immunogenic peptide pools for eliciting CD8 T cell-mediated clearance of HIV-infected cells. This study will accelerate the development of a new HIV treatment strategy that combines Nef blockade for MHC-I restoration and immunotherapies that elicit effective CTL response and provide the evidentiary basis for progressing to an in vivo model before this approach can be applied in a clinical setting.