Impact of HSV-2 shedding on the vaginal microbiome in the female genital tract

HSV-2 脱落对女性生殖道阴道微生物组的影响

基本信息

批准号：
9308822
负责人：
CHRISTINE MICHELLE JOHNSTON
金额：
$ 23.2万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/9308822
关键词：
Acyclovir Address Antiviral Agents Bacteria Bacterial Vaginosis Biological Biological Assay CCR5 gene CD4 Positive T Lymphocytes CD8-Positive T-Lymphocytes Cervix Uteri Chronic Clinical Communities Complex Coupled Data Detection Development Diagnosis Disease Economic Burden Enrollment Epidemic Epidemiology Female Frequencies Genital system Goals HIV HIV Infections HIV Seronegativity High-Throughput Nucleotide Sequencing Human Herpesvirus 2 Immune Immune Evasion Immune response Immunologics Infection Inflammation Inflammatory Inflammatory Response Interferon Type II Interleukin-1 beta Intervention Knowledge Lactobacillus Lesion Link Maintenance Measures Mediating Methods Molecular Morbidity - disease rate Mucous Membrane Natural History Nature Participant Phenotype Proteins Recording of previous events Recurrence Reproductive Health Research Risk Risk Factors SLPI gene Sampling Sexually Transmitted Diseases Simplexvirus Site Skin Specimen Surface Swab T-Lymphocyte Time Tissues Universities Vagina Virus Washington Woman antileukoprotease cytokine experience genital herpes genital secretion improved inflammatory marker innovation insight interest microbial microbiome microbiota mortality novel pathogen pyrosequencing rRNA Genes reproductive tract response risk sharing routine Bacterial stain sample collection seropositive therapy design vaginal fluid vaginal microbiome

项目摘要

Project Summary Herpes simplex virus type 2 (HSV-2) and bacterial vaginosis (BV) are two of the most prevalent infections of the female genital tract. Both infections are associated increased risk for HIV acquisition. While HSV-2 and BV have been epidemiologically linked in many studies, the direction and mechanism of their interaction is not known. Our group has pioneered daily participant-collected sampling to show that HSV-2 is detected ("shed") frequently, on 10-28% of days, from genital surfaces, and that HSV-2 shedding is asymptomatic the majority of the time. More recent data has demonstrated that HSV-2 genital lesions and asymptomatic shedding episodes are associated with a persistent inflammatory cellular response, with IFN-γ producing HSV-specific CD4+ and tissue resident CD8+ T-cells. These data suggest that HSV-2 infection chronically and fundamentally alters inflammation the genital tract. Drs. Marrazzo and Fredricks have similarly used frequent vaginal sampling and molecular methods to document rapid shifts in the vaginal microbiome, and to discover novel BV-associated bacteria (BVAB). This proposal seeks to elucidate the temporal and mechanistic nature of the HSV-2/BV association and to identify the components of the inflammatory response to HSV-2 that may increase the risk of BV. We hypothesize that frequent HSV-2 genital shedding causes increased levels of inflammatory cytokines, such as IL-1ß and IFN-γ, and decreased levels of antiviral innate immune molecules (i.e. secretory leukocyte protease inhibitor, SLPI) and that these changes result in fluctuations in the vaginal microbiome which predispose women to develop BV. To address these hypotheses, we will enroll 40 HSV-2 seropositive, HIV seronegative women with a history of BV in the past year who will self-collect genital and vaginal specimens daily for 30 days. We will also enroll 20 HSV-2 seropositive women without a clinical history of BV. We will quantify HSV using a validated PCR assay, and we will measure changes in the microbiome using gram stain (Nugent score), quantitative PCR (qPCR) and broad range 16S rRNA gene PCR with high throughput sequencing during HSV shedding episodes. Daily vaginal levels of innate immune proteins and cytokines associated with HSV-2 shedding will be measured. Relationships between HSV-2 shedding and Nugent score, quantity and presence of BVAB, and cytokine levels will be determined. These women will then be placed on suppressive acyclovir, which decreases HSV-2 shedding by 70-80%. We will determine whether suppression of HSV-2 shedding results in improved vaginal flora as measured by a decrease in Nugent score and increased concentration of lactobacilli. This integrated proposal will determine the relationship between HSV-2 shedding and the vaginal microbiome, and will give insight into the mechanistic basis of the association to provide a framework for appreciating how these infections influence inflammation within the genital tract. Ultimately, this proposal will contribute vital and novel information to the design of interventions that can modulate BV, and will provide us with knowledge to better promote women's reproductive health.

项目概要单纯疱疹病毒 2 型 (HSV-2) 和细菌性阴道病 (BV) 是两种最常见的感染女性生殖道感染会增加感染 HIV 的风险。 BV在许多研究中已被流行病学联系起来，但它们相互作用的方向和机制尚不清楚。我们的团队率先每天对参与者进行采样，以显示检测到 HSV-2（“脱落”）。 10-28% 的日子里，频繁地从生殖器表面传播，并且 HSV-2 的脱落大多数是无症状的最近的数据表明，HSV-2 生殖器病变和无症状脱落事件。与持续的炎症细胞反应有关，其中 IFN-γ 产生 HSV 特异性 CD4+ 和这些数据表明 HSV-2 感染会长期且从根本上改变。 Marrazzo 和 Fredricks 博士也同样采用了频繁的阴道取样方法。分子方法记录阴道微生物群的快速变化，并发现新的 BV 相关该提案旨在阐明 HSV-2/BV 的时间和机制性质。关联并确定可能增加风险的 HSV-2 炎症反应的组成部分我们认为频繁的 HSV-2 生殖器脱落会导致炎症水平增加。细胞因子，如 IL-1ß 和 IFN-γ，以及抗病毒先天免疫分子（即分泌性免疫分子）水平降低白细胞蛋白酶抑制剂，SLPI），这些变化导致阴道微生物群的波动为了验证这些假设，我们将招募 40 名 HSV-2 血清阳性的患者，过去一年内有 BV 病史的 HIV 血清阴性女性将自行收集生殖器和阴道我们还将招募 20 名没有 BV 临床病史的 HSV-2 血清阳性女性，持续 30 天。我们将使用经过验证的 PCR 检测来量化 HSV，并且我们将使用以下方法来测量微生物组的变化革兰氏染色（Nugent 评分）、定量 PCR (qPCR) 和广泛的 16S rRNA 基因 PCR，具有高 HSV 脱落期间的每日阴道先天免疫蛋白水平和通量测序。将测量与 HSV-2 脱落相关的细胞因子和 HSV-2 脱落之间的关系。然后将确定 Nugent 评分、BVAB 的数量和存在以及细胞因子水平。服用抑制性阿昔洛韦，可减少 70-80% 的 HSV-2 脱落。抑制 HSV-2 脱落可改善阴道菌群（通过 Nugent 评分降低来衡量）和乳酸菌浓度增加这一综合建议将确定两者之间的关系。 HSV-2 脱落和阴道微生物组，并将深入了解这种关联的机制基础提供一个框架来了解这些感染如何影响生殖道内的炎症。最终，该提案将为干预措施的设计提供重要且新颖的信息，这些信息可以调节BV，将为我们更好地促进女性生殖健康提供知识。