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：该提案支持为萨博博士提供为期五年的培训计划，以获得她所需的技能 成为一名独立的内科科学家和生殖免疫学专家。萨博医生有一个 具有基础科学和免疫学背景，并计划将重点转向面向临床的翻译 研究。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 来帮助她完成这一转变。研究计划：细菌性阴道病(BV)和次优阴道细菌分类群 与艾滋病毒感染有关。发生这种情况的确切机制尚不清楚，但可能 与宫颈炎症加重有关。树突状细胞(DC)是调节炎症反应的关键细胞。 粘膜组织的状态，并可能在生殖器感染艾滋病毒方面发挥作用。然而，DC保持最低限度的 以宫颈为特征的。这项建议的目的是检查次佳的阴道细菌和 阴道细菌物种多样性的增加与宫颈DC总数的增加有关。我们有 提出三点旨在回答这一问题。对于目标1和目标2，我们将进行横断面研究 在肯尼亚蒙巴萨，女性性工作者(FSWs)参加了一项纵向开放队列研究。阴道拭子 宫颈活检样本将在一次访问中收集。以确定高危细菌分类群是否 随着DC数量的增加(AIM 1)，我们将评估浓度与 阴道细菌(用定量聚合酶链式反应[qPCR]测量)和宫颈DC总数(用流量测量 来自宫颈活检组织消化的细胞术)。检查阴道细菌与细菌之间的关系 物种多样性和宫颈DC(AIM 2)，我们将进行广泛的聚合酶链式反应和高通量测序 以计算每个患者的香农多样性指数(SDI)，并将SDI进行比较 用流式细胞仪检测DC的数量。在目标3中，我们将检验BV治疗的假设 减少宫颈DC的数量。作为探索性结果，DC的激活将通过测量进行评估 细胞表面共刺激标记物和细胞内促炎细胞因子用于所有AIMS。加在一起，这些 AIMS有可能为次优阴道微生物区系之间的机械联系提供证据， 宫颈炎症和女性对艾滋病毒的易感性。培训计划：培训的六个核心学习目标 K23获奖期包括流行病学、临床研究、全球卫生、高维度数据分析、 粘膜免疫学和阴道微生物群。选择这些目标是为了确保萨博博士获得 领导自己的临床研究并开始独立研究生涯所需的技能 旨在阐明生殖免疫系统在健康和疾病中的作用的翻译研究。培训以达到 完成六个主要学习目标将通过指导、课程作业、 实施拟议的研究计划、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. .