Genital Anaerobes, Inflammation and HIV Risk; Rakai

描述（由申请人提供）：“经典”性传播感染（STI）和细菌性阴道病（BV）与HIV易感性和传染性相关。然而，为预防艾滋病毒而进行的性传播感染控制试验基本上是消极的。我们假设其他生殖器微生物可能会引起炎症，并导致艾滋病毒的风险增加。我们将使用男性包皮环切术（MC）试验期间收集的数据和样本来评估这一假设。HIV阴性的男性被随机分配到立即MC（干预）或MC延迟2年（对照）。男性在0、6、12和24个月时提供血液和阴茎拭子。包皮在MC时被保存。这些男性的女性伴侣在0、12和24个月时提供了血液和阴道拭子。使用这些材料，我们将解决以下目标：目标1。1a）评估100名干预和100名对照男性及其200名女性伴侣的生殖器厌氧菌负荷与生殖器细胞因子/趋化因子水平之间的相关性。1.b）评估这些暴露与包皮组织中组织学炎症之间的关联。1.c.使用目标1.a中的数据，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.评估生殖器厌氧菌的负担和与艾滋病毒获得相关的细胞因子/趋化因子水平，使用病例对照分析105名男性和89名女性艾滋病毒血清转换者，以及388名艾滋病毒未感染的对照。将通过多变量logistic回归估计调整后的比值比。目标3。在310对HIV不一致夫妇中，我们将估计与生殖器厌氧菌负担和细胞因子/趋化因子水平相关的HIV传播/获得率。将通过Poisson多变量回归估计HIV感染/传播的发生率比。在另一个目标4中，我们将利用目标1-3的数据评估厌氧菌和炎症对MC预防男性HIV疗效的介导作用。实验室方法：将通过16 S rRNA qPCR和基于16 SrRNA基因的焦磷酸测序评估总细菌负荷以及相对和绝对厌氧菌负荷。将使用多重夹心ELISA（SearchLight，Aushon Biosystems，MA）测定TNF α、IFN？、IL-1、IL-8、IL-6、IL-12、RANTES、IFN诱导的单核因子？(MIG/CXCL 9）、巨噬细胞趋化蛋白（MCP 1/CCL 2）和干扰素诱导蛋白-10（IP 10/CXCL 10）。 这是流行病学家，分子生物学家和免疫学家之间的多学科合作，并包括一个新的研究者（博士L价格，共同PI）。 公共卫生相关性：如果生殖器厌氧菌引起粘膜炎症并与HIV获得/传播相关，则该研究可以通过控制促炎性生殖器厌氧菌（例如，通过使用局部防腐剂或抗生素），或调节男性和女性生殖器炎症反应的试剂。这可以为杀微生物剂的开发提供信息。对粘膜免疫学的深入了解也可能有助于未来粘膜HIV疫苗的开发。

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.