The impact of vaginal microbiota on cervical dendritic cells: an observational study of women from sub-Saharan Africa at high risk for HIV acquisition.

摘要 摘要：该提案支持Sabo博士的五年培训计划，以获得她所需的技能， 成为一名独立的医学科学家和生殖免疫学专家。萨博博士有一个 在基础科学和免疫学的背景，并计划将她的重点转移到临床导向，翻译 research.她制定了一套与她的研究兴趣相一致的目标，以及六个核心学习目标。 帮她完成这个转变研究计划：细菌性阴道病（BV）和次优阴道细菌分类群 与艾滋病感染有关。发生这种情况的确切机制尚不清楚，但可能 与宫颈炎症增加有关。树突状细胞（DC）在调节炎症反应中起关键作用。 粘膜组织的状态，并可能在生殖器艾滋病毒的获得中发挥作用。然而，发展中国家仍然很少 以子宫颈为特征。这项建议的目的是检查次优阴道细菌和 增加的阴道细菌物种多样性与增加的总宫颈DC相关。我们有 提出了三个目的来回答这个问题。对于目标1和2，我们将进行一项横断面研究， 女性性工作者（FSW）参加了一项在肯尼亚蒙巴萨进行的纵向开放队列研究。阴道拭子 并在单次访视时采集宫颈活检样本。为了确定高风险细菌分类群是否 与DC数量增加（AIM 1）相关，我们将评估浓度之间的关联， 阴道细菌（通过定量PCR [qPCR]测量）和宫颈DC的总数（通过流式细胞仪测量） 来自宫颈活检组织切片的细胞计数）。为了检测阴道细菌与 物种多样性和宫颈DC（AIM 2），我们将进行大范围PCR和高通量测序 计算每例患者的Shannon多样性指数（SDI），并将SDI进行比较 与通过流式细胞术分离的DC的数量有关。在AIM 3中，我们将检验BV治疗 减少宫颈DC的数量。作为探索性结局，将通过测量评估DC激活 细胞表面共刺激标志物和细胞内促炎细胞因子的所有目的。所有这些 目的有可能为次优阴道微生物群之间的机械联系提供证据， 宫颈炎和女性艾滋病易感性。培训计划：六个核心学习目标 K23奖学金期间包括流行病学，临床研究，全球健康，高维数据分析， 粘膜免疫学和阴道微生物组。选择这些目标是为了确保Sabo博士获得 领导自己的临床研究和开展独立研究生涯所需的技能， 翻译研究，以阐明生殖免疫系统在健康和疾病中的作用。培训 完成六个关键的学习目标将通过指导，课程， 执行拟议的研究计划、举办地方研讨会以及国家和国际会议。 .

ABSTRACT Summary: This proposal supports a five-year training program for Dr. Sabo to gain the skills she needs to become an independent physician-scientist and an expert in reproductive immunology. Dr. Sabo has a background in basic science and immunology, and plans to shift her focus to clinically-oriented, translational research. She has developed a set of aims that align with her research interests, and six core learning objectives to help her make this transition. Research plan: Bacterial vaginosis (BV) and sub-optimal vaginal bacterial taxa have been associated with HIV acquisition. The precise mechanism by which this occurs is unknown, but may be related to increased cervical inflammation. Dendritic cells (DCs) are critical for regulating the inflammatory state of mucosal tissues, and likely play a role in genital acquisition of HIV. However, DCs remain minimally characterized in the cervix. The purpose of this proposal is to examine if sub-optimal vaginal bacteria and increased vaginal bacterial species diversity are associated with increased total cervical DCs. We have proposed three aims to answer this question. For AIMS 1 and 2, we will perform a cross sectional study of female sex workers (FSWs) enrolled in a longitudinal, open cohort study in Mombasa, Kenya. Vaginal swabs and cervical biopsy samples will be collected at a single visit. To determine if high-risk bacterial taxa are associated with increased DC number (AIM 1), we will evaluate the association between concentrations of vaginal bacteria (measured by quantitative PCR [qPCR]) and total numbers of cervical DCs (measured by flow cytometry from tissue digests of cervical biopsies). To examine the relationship between vaginal bacterial species diversity and cervical DCs (AIM 2), we will perform broad range PCR with high throughput sequencing of vaginal swabs to calculate the Shannon Diversity Index (SDI) for each patient, and the SDI will be compared to the number of DCs isolated by flow cytometry. In AIM 3, we will test the hypothesis that treatment of BV reduces the number of cervical DCs. As an exploratory outcome, DC activation will be assessed by measurement of cell surface co-stimulatory markers and intracellular pro-inflammatory cytokines for all aims. Together, these aims have the potential to provide evidence for a mechanistic link between sub-optimal vaginal microbiota, cervical inflammation, and HIV susceptibility in women. Training plan: The six core learning objectives for this K23 award period include training in epidemiology, clinical studies, global health, high dimensional data analysis, mucosal immunology, and the vaginal microbiome. These goals were selected to ensure that Dr. Sabo acquires the skills necessary to lead her own clinical studies and launch an independent research career performing translational studies to elucidate the role of the reproductive immune system in health and disease. Training to complete the six key learning objectives will be achieved through a combination of mentorship, coursework, implementation of the proposed research plan, local seminars, and national and international meetings. .