Enhancement of HIV transmission by hormones and bacterial metabolites

描述（由申请人提供）：本提案旨在提高我们对激素水平和生殖道微生物变化如何影响女性感染艾滋病毒风险的理解。这些都是重要的研究领域，因为一半以上新的艾滋病毒感染是妇女获得的，主要是通过异性传播。本研究的目的是确定正常月经周期（内源性激素）、DMPA避孕药（合成黄体酮）和细菌性阴道病期间的激素变化如何影响宫颈外CD4+ T淋巴细胞的蛋白表达及其对HIV融合和生产性感染的易感性。这些研究有助于确定激素效应、微生物群效应以及激素与阴道微生物之间的相互作用效应。我们发现，细菌代谢物短链脂肪酸（SFCAs）存在于健康的宫颈阴道液中，并在BV期间急剧增加，通过抑制组蛋白去乙酰化酶（hdac），强烈提高CD4+ T细胞对生产性HIV感染的易感性。最后，我们将研究一种通过使细胞对HIV感染更有抵抗力来减少HIV传播的新策略。本项目要调查的三个重叠区域是：1。测定激素水平和BV对宫颈外活检CD4+ T淋巴细胞易感性和激活状态的影响。使用我们的联合报告病毒系统，流式细胞术将直接测量宫颈外CD4+ T细胞对HIV融合和生产性感染的易感性。CD4+ T细胞的激活状态将通过流式细胞术和无标记蛋白质组学进行量化，这是一种确定特定激活途径的公正而强大的策略。先进的生物信息学将用于识别有助于子宫颈外CD4+ T细胞中HIV复制的特定激活途径。2. 量化存在于宫颈阴道灌洗（CVL）样品中的细菌种类和代谢物，并将代谢物与特定种类或细菌群相关联。CVL样本将被分析细菌种类和细菌代谢物——包括短链脂肪酸（SCFAs），它存在于健康女性生殖道中，在细菌性阴道炎中升高。我们最近发现SCFAs通过抑制hdac的机制增强HIV对CD4+ T细胞的感染。先进的生物信息学将用于确定细菌及其产物与宫颈外CD4+ T细胞蛋白表达和HIV易感性之间的相关性。3. 确定garcinol在不同激素模式和宫颈阴道微生物群的女性体内降低宫颈外CD4+ T细胞对HIV易感性的能力。Garcinol，一种组蛋白乙酰转移酶抑制剂，可以逆转

DESCRIPTION (provided by applicant): This proposal aims to improve our understanding of how variations in hormone levels and reproductive tract microbes contribute to HIV acquisition risk in women. These are significant areas of research because more than half of new HIV infections are acquired by women, predominantly through heterosexual transmission. The goal of the proposed research project is to define how hormonal variation during the normal menstrual cycle (endogenous hormones) and with DMPA contraceptives (synthetic progestin) and bacterial vaginosis affect protein expression in ectocervical CD4+ T lymphocytes and their susceptibility to fusion and productive infection with HIV. These studies are powered to allow determination of hormonal effects, microbiota effects and interaction effects between hormones and vaginal microbes. We have found that bacterial metabolites called short chain fatty acids (SFCAs) that are present in healthy cervicovaginal fluid and increase dramatically during BV, strongly heighten the susceptibility of CD4+ T cells to productive HIV infection through the inhibition of histone deacetylases (HDACs). Finally, we will investigate a novel strategy to reduce HIV transmission by making cells more resistant to HIV infection. The three overlapping areas to be investigated in this project are: 1. Determine the effects of hormone levels and BV on susceptibility and activation state of CD4+ T lymphocytes from ectocervical biopsies. Ectocervical CD4+ T cell susceptibility to fusion and productive infection by HIV will be directly measured using our combination reporter virus system by flow cytometry. Activation states of CD4+ T cells will be quantified by flow cytometry and label-free proteomics, an unbiased and powerful strategy for identifying specific activation pathways. Advanced bioinformatics will be used to identify specific activation pathways that contribute to HIV replication in ectocervical CD4+ T cells. 2. Quantify bacterial species and metabolites present in cervicovaginal lavage (CVL) samples and correlate metabolites with specific species or groups of bacteria. CVL samples will be analyzed for bacterial species and for bacterial metabolites-including short chain fatty acids (SCFAs) that are present in the healthy female reproductive tract and elevated in BV. We have recently found that SCFAs enhance HIV infection of CD4+ T cells via a mechanism involving inhibition of HDACs. Advanced bioinformatics will be used to identify correlations between bacteria and their products and ectocervical CD4+ T cell protein expression and susceptibility to HIV. 3. Determine the ability of garcinol to reduce the susceptibility of ectocervical CD4+ T cells to HIV ex vivo in women with different hormone patterns and cervicovaginal microbiota. Garcinol, a histone acetyltransferase inhibitor, reverses the effects of SCFA-mediated HDAC inhibition of HIV susceptibility in vitro, and actually reduces infection even in unstimulated cells. These findings suggest that garcinol, or similar compounds, may represent a novel strategy to reduce heterosexual transmission of HIV by inhibiting productive infection of ectocervical CD4+ T cells.