Mechanisms of Resistance in HIV-1 Exposed Sero-negative IV-drug Users: Induction of IFN-mediated Factors and S100 Proteins as Determinants of NK Cell-Mediated Clearance and Low CD4+ T Cell Infectivity

HIV-1 血清阴性静脉注射药物使用者的耐药机制：诱导 IFN 介导的因子和 S100 蛋白作为 NK 细胞介导的清除和低 CD4 T 细胞感染性的决定因素

• 批准号: 9248331
• 负责人: Costin Tomescu
• 金额: $ 22.74万
• 依托单位: WISTAR INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2019-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9248331
• 关键词: Autologous; Biological Assay; CD4 Positive T Lymphocytes; Cell Degranulation; Cell physiology; Cells; Cytoplasmic Protein; Data; Drug Addiction; Drug usage; Environment; Exhibits; Family; Goals; Granzyme; HIV; HIV-1; Host resistance; Immune; In Vitro; Individual; Infection; Interferons; Lead; Link; MHC Class I Genes; Measures; Mediating; Modeling; NK Cell Activation; Natural Increases; Natural Killer Cells; Needle Sharing; Needle-Exchange Programs; Pathway interactions; Philadelphia; Proteins; Proteome; Proteomics; Recruitment Activity; Reporting; Research; Resistance; Resistance to infection; Risk; Role; S100 Proteins; S100A4 gene; STAT1 gene; Small Interfering RNA; T-Lymphocyte; Testing; Viral; Virus; autocrine; cohort; high risk; high risk behavior; immunological synapse; immunoregulation; in vivo; innovation; intravenous drug user; knock-down; link protein; member; novel; proteomic signature; public health relevance; receptor; resistance mechanism

 DESCRIPTION (provided by applicant): The absence of clear immune correlates for protection against HIV-1 highlight the critical need to identify new pathways of host-resistance from infection. The overall goal of this R21 proposal is to identify novel mechanism(s) of protection in a cohort of HIV-exposed individuals who remain sero-negative (HESN) despite many years of high-risk behavior and exposure. Previous studies of HESN subjects exposed to HIV-1 through IV-drug use and needle-sharing (HESN-IDU) have identified several potential innate and intrinsic mechanisms of protection, including heightened Natural Killer (NK) cell function and increased resistance of CD4+ T cells to HIV-1 infection. Our preliminary data now provide the basis to test an innovative model for how these innate and intrinsic mechanisms of resistance may cooperate to provide a sustained barrier against HIV-1 infection in HESN-IDU subjects. Using a well-described cohort of high-risk HESN-IDU subjects from Philadelphia, we have identified a unique proteomic signature on NK cells from HESN-IDU subjects including the elevated expression of multiple interferon-induced proteins such as ISG-15, MHC-Class I, Granzyme, STAT1/2, as well as the increased expression of several members of the S100 family of immuno-modulatory proteins not previously identified in HESN subjects. Specifically, we identified the increased expression of two cytoplasmic S100 proteins, S100A4 and S100A6, that may stimulate NK degranulation capacity against virally infected cells and a secreted S100 protein, S100A14, that may augment HIV-1 resistance in CD4+ T cells. These results indicate that high-risk needle sharing in protected HESN-IDU subjects may trigger an anti-viral environment involving secreted Interferon and/or S100 proteins that can lead to greater NK activity against virally infected cells and increased CD4+ target cell resistance to HIV-1. In Specific Aim 1, we will measure the ability of NK cells from HESN-IDU subjects and NS-IDU controls to limit the replication capacity of Autologous HIV-1 infected CD4+ primary T cells using an HIV Suppression Assay. We will also investigate if NK cells from HESN-IDU subjects possess increased CD107a degranulation against HIV-1 infected heterologous SupT1 cells and if this correlates with enhanced S100A4 and S100A6 recruitment into the immunological synapse. In Specific Aim 2, we will test the resistance of purified CD4+ T cells from HESN-IDU subjects to HIV-1 infection and investigate the ability of the secreted S100A14 protein to further limit HIV-1 replication capacity. We will investigate the expression of known and uncharacterized host restriction factors in CD4+ T cells from HESN-IDU subjects by proteome analysis and correlate them with infectivity. Together, the Specific Aims proposed in this R21 will test the novel hypothesis that increased expression of interferon-induced factors and S100 proteins in HESN-IDU subjects augment innate and intrinsic mechanisms of resistance by increasing NK-mediated clearance of virally infected cells and reducing the efficiency of HIV-1 replication in CD4+ T cells.

 描述（由适用提供）：缺乏明确的免疫力来防止HIV-1突出显示识别感染宿主抵抗的新途径的关键需求。该R21提案的总体目标是在艾滋病毒暴露的个体中确定新型的保护机制，这些人保持血清阴性（HESN）的高风险行为和暴露。先前对通过IV-Crug使用和共享针刺（HESN-IDU）暴露于HIV-1受试者的HESN受试者的研究已经确定了几种潜在的先天和内在保护机制，包括增强的天然杀伤（NK）细胞功能以及CD4+ T细胞对HIV-1感染的耐药性的提高。现在，我们的初步数据为测试这些抗药性的先天和内在机制如何协调以提供针对HIV-1感染HES-IDU受试者的持续障碍的创新模型提供了基础。 Using a well-described cohort of high-risk HESN-IDU subjects from Philadelphia, we have identified a unique proteinomic signature on NK cells from HESN-IDU subjects including the elevated expression of multiple interferon-induced proteins such as ISG-15, MHC-Class I, Granzyme, STAT1/2, as well as the increased expression of several members of the S100 family of immuno-modulatory以前在HESN受试者中未鉴定的蛋白质。具体而言，我们确定了两种细胞质S100蛋白S100A4和S100A6的表达增加，这可能会刺激针对实际感染的细胞的NK脱粒能力和一个分泌的S100蛋白S100A14，这些蛋白可能会增强CD4+ T细胞中HIV-1的耐药性。这些结果表明，受保护的HESN-IDU受试者中的高风险针共享可能会触发涉及分泌干扰素和/或S100蛋白的抗病毒环境，这些蛋白可能导致对实际上感染细胞的NK活性更大，并增加CD4+靶细胞对HIV-1的抗性。在特定的目标1中，我们将使用HIV抑制测定法测量NK细胞和NS-IDU对照中NK细胞的能力限制自体HIV-1感染CD4+原代T细胞的复制能力。我们还将研究HESN-IDU受试者的NK细胞是否对HIV-1感染的异源SUPT1细胞具有增加的CD107A脱粒化，并且这是否与增强的S100A4和S100A6募集到免疫突触中。在特定的目标2中，我们将测试纯化的CD4+ T细胞从HESN-IDU受试者对HIV-1感染的抗性，并研究分泌的S100A14蛋白进一步限制HIV-1复制能力的能力。我们将通过蛋白质组分析研究来自HESN-IDU受试者的CD4+ T细胞中已知和未表征的宿主限制因子的表达，并将其与感染相关联。总之，在本R21中提出的具体目的将测试新的假设，即在HESN-IDU受试者中增加了干扰素诱导的因子和S100蛋白的表达，从而增强了先天性和内在的抗药性机制，通过增加NK介导的差异的清除率，从而增加了对实际感染的细胞的清除，并减少了CD4+ T细胞中HIV-1复制的含量。