Genital Anaerobes, Inflammation and HIV Risk; Rakai

描述（由申请人提供）：“经典”性传播感染（STIs）和细菌性阴道病（BV）与HIV易感性和传染性有关。然而，控制性传播感染预防艾滋病毒的试验基本上是阴性的。我们假设，其他生殖器官可能会引起炎症，并增加艾滋病毒的风险。我们将使用在男性包皮环切术（MC）试验中收集的数据和样本来评估这一假设。hiv阴性男性随机分为立即MC组（干预组）和延迟2年MC组（对照组）。男性在0、6、12和24个月时提供血液和阴茎拭子。在MC时保存包皮。这些男性的女性伴侣在0、12和24个月时提供血液和阴道拭子。使用这些材料，我们将解决以下目标：1a)评估100名干预组和100名对照组男性及其200名女性伴侣中厌氧菌的生殖负担与生殖细胞因子/趋化因子水平之间的相关性。b)评估这些暴露与包皮组织组织学炎症之间的关系。1.摄氏度。使用Aim 1的数据。a，我们将评估MC对干预组与对照组男性及其伴侣的生殖器厌氧菌和炎症标志物的影响。目标2。通过对105名男性和89名女性艾滋病毒感染者以及388名未感染艾滋病毒的对照组进行病例对照分析，评估生殖器厌氧菌负担和与艾滋病毒感染相关的细胞因子/趋化因子水平。调整后的优势比将通过多变量逻辑回归估计。目标3。在310对HIV不一致的夫妇中，我们将估计HIV传播/获得率与生殖器厌氧菌负担和细胞因子/趋化因子水平的关系。HIV感染/传播的发生率比值将通过泊松多变量回归估计。在另一个目标4中，我们将利用目标1-3的数据来评估厌氧菌和炎症对MC预防男性HIV疗效的中介作用。实验室方法：通过16SrRNA qPCR和16SrRNA基因焦磷酸测序评估细菌总负荷、相对和绝对厌氧菌负荷。细胞因子和趋化因子浓度将使用多重夹心ELISA （SearchLight, Aushon Biosystems， MA）检测TNFa、IFN？， IL-1, IL-8, IL-6, IL-12， RANTES， IFN诱导的单因子？（MIG/CXCL9）、巨噬细胞趋化蛋白（MCP1/CCL2）和干扰素诱导蛋白-10 （IP10/CXCL10）。

DESCRIPTION (provided by applicant): "Classical" sexually transmitted infections (STIs) and bacterial vaginosis (BV) are associated with HIV susceptibility and infectivity. However, trials of STI control for HIV prevention have been largely negative. We hypothesize that other genital organisms may induce inflammation and contribute to increased risk of HIV. We will use data and samples collected during a trial of male circumcision (MC) to assess this hypothesis. HIV-negative men were randomized to immediate MC (intervention) or MC delayed by 2 years (controls). Men provided blood and penile swabs at 0, 6, 12 and 24 months. Foreskins were preserved at time of MC. Female partners of these men provided blood and vaginal swabs at 0, 12 and 24 months. Using these materials, we will address the following aims: Aim1. 1a) Assess the correlation between the genital burden of anaerobes and levels of genital cytokines/chemokines in 100 intervention and 100 control men and their 200 female partners.1.b) Assess the association between these exposures and histologic inflammation in foreskin tissues. 1.c. Using the data from Aim 1.a, we will assess the effects of MC on genital anaerobes and inflammatory markers in intervention versus control arm men and their partners. Aim 2. Assess the burden of genital anaerobes and levels of cytokines/chemokines associated with HIV acquisition using a case-control analysis of 105 male and 89 female HIV seroconverters, and 388 HIV-uninfected controls. Adjusted odds ratios will be estimated by multivariable logistic regression. Aim 3. In 310 HIV discordant couples we will estimate rates of HIV transmission/acquisition associated with genital anaerobe burden and cytokine/ chemokine levels. Incidence rate ratios of HIV acquisition/transmission will be estimated by Poisson multivariable regression. In an additional Aim 4, we will utilize the data from Aims 1-3 to assess the mediating role of anaerobes and inflammation on MC efficacy for HIV prevention in men. Laboratory methods: Total bacterial load, and the relative and absolute anaerobe burdens will be assessed by16S rRNA qPCR and 16SrRNA gene-based pyrosequencing. Cytokine and chemokine concentrations will be assayed using multiplex sandwich ELISA (SearchLight, Aushon Biosystems, MA) for TNFa, IFN?, IL-1¿, IL-8, IL-6, IL-12, RANTES, Monokine Induced by IFN? (MIG/CXCL9), Macrophage Chemotactic Protein (MCP1/CCL2), and Interferon Inducible Protein-10 (IP10/CXCL10). This is a multidisciplinary collaboration between epidemiologists, molecular biologists and immunologists, and includes a new investigator (Dr. L Price, co-PI). PUBLIC HEALTH RELEVANCE: If genital anaerobes cause mucosal inflammation and are associated with HIV acquisition/transmission, the study could facilitate development of novel approaches to HIV prevention by control of pro-inflammatory genital anaerobes (e.g., by use of topical antiseptics or antibiotics), or agents to modulate genital inflammatory responses in men and women. This could inform microbicide development. Insights into mucosal immunology may also contribute to development of future mucosal HIV vaccines.