The Role of Uncultivated Bacteria in Bacterial Vaginosis

描述（由申请人提供）：人类阴道的微生物群可以深刻影响妇女和新生儿的健康。例如，患有细菌性阴道病（BV）的妇女获得性传播感染（如艾滋病毒）、患上盆腔炎和早产的风险增加。无论是考虑到每年数百万因阴道分泌物就诊的直接费用，还是考虑到照顾早产儿等间接费用，与细菌性阴道炎相关的医疗费用都是巨大的。尽管最近的进展已经使用分子方法（PCR）来更好地描述细菌性阴道炎的微生物学，但对于为什么有些妇女会出现这种情况，人们缺乏了解。女性发生细菌性阴道炎后阴道微生物群的变化已被很好地描述，但细菌性阴道炎的微生物、生理和行为前因却知之甚少。这项建议旨在通过一项纵向研究来收集有感染细菌性阴阳炎风险的妇女的数据，从而克服这一知识差距。广泛范围16S rRNA基因PCR与焦磷酸测序和细菌特异性定量PCR将用于表征阴道微生物。研究导致细菌性阴道炎的因素对了解其发病机制至关重要。这些知识可用于改进诊断、预防和治疗方法。虽然可以使用甲硝唑和克林霉素等抗生素治疗细菌性肠胃炎，但复发是常见的，3个月复发率高达30%，1年复发率为50%。复发率如此之高的原因尚不清楚。高复发率使改善细菌性阴道炎不良健康后果的努力复杂化。这一建议旨在通过研究阴道外可能允许阴道播种的细菌库来描述与细菌性阴道炎复发相关的微生物因素。在抗生素治疗期间和之后，将对感染细菌性阴道炎的妇女（以及一部分性伴侣）进行随访，使用聚合酶链反应评估阴道和阴道外部位的阴道细菌浓度。预计阴道外蓄水池的定植将是BV复发的主要危险因素。生物膜在细菌性肠胃炎的发病机制中是否重要仍在争论中，基于一些小型横断面研究的有趣数据支持了这一概念。荧光原位杂交（FISH）将用于评估从患有和不患有细菌性阴道炎的妇女获得的阴道样本中细菌生物膜的患病率。这里将研究非活检样本的价值。抗生素治疗后将评估阴道细菌生物膜的变化，以确定细菌性阴道炎的复发是否与生物膜群落中细菌的持续存在有关。如果生物膜在细菌性胃炎中发挥重要作用，这将为基于破坏生物膜形成和细胞信号传导的治疗和预防开辟许多新的途径。

DESCRIPTION (provided by applicant): The microbiota of the human vagina can profoundly affect the health of women and neonates. For instance, women with the condition bacterial vaginosis (BV) have increased risks of acquiring sexually transmitted infections such as HIV, developing pelvic inflammatory disease, and experiencing preterm birth. Healthcare costs associated with BV are enormous, whether considering the direct costs related to the millions of healthcare visits annually for vaginal discharge, or the indirect costs such as care of pre-term infants. Despite recent advances that have used molecular methods (PCR) to better describe the microbiology of BV, there is a lack of understanding regarding why some women develop this condition. Changes in the vaginal microbiota after women develop BV are well described, but the microbiological, physiological, and behavioral antecedents to BV are poorly understood and connected. This proposal seeks to overcome this knowledge gap by collecting data on women at risk for BV in a longitudinal study designed to capture incident and relapsing cases. Broad range 16S rRNA gene PCR with pyrosequencing and bacterium-specific quantitative PCR will be used to characterize vaginal microbiotas. Changes in vaginal bacterial communities will be linked to physiological factors (menses), medication use (hormonal contraceptives, antibiotics), and behavioral factors (sexual practices, condom use). Studying the factors that lead to BV is critical for understanding pathogenesis. This knowledge can be used to develop improved methods for diagnosis, prevention, and treatment. Although antibiotics such as metronidazole and clindamycin can be used to treat BV, recurrence is common, with recurrence rates of up to 30% at 3 months and >50% at one year. The reasons for this high rate of recurrence are not known. High relapse rates complicate efforts to ameliorate the adverse health consequences of BV. This proposal seeks to describe the microbiological factors associated with recurrence of BV by studying extra-vaginal reservoirs of bacteria that may allow seeding of the vagina. Women with BV (and a subset of sex partners) will be followed during and after antibiotic treatment to assess concentrations of vaginal bacteria in vaginal and extra-vaginal sites using PCR. It is anticipated that colonization of extra-vaginal reservoirs will be a major risk factor for recurrence of BV. There is ongoing debate whether biofilms are important in the pathogenesis of BV, with intriguing data supporting this concept based on some small cross-sectional studies. Fluorescence in situ hybridization (FISH) will be used to assess the prevalence of bacterial biofilms in vaginal samples obtained from women with and without BV. The value of non-biopsy samples will be studied here. Changes in vaginal bacterial biofilms will be assessed after antibiotic treatment to determine if recurrence of BV is associated with persistence of bacteria in biofilm communities. If biofilms are shown to be important in BV, this would open many new avenues for treatment and prevention based on disruption of biofilm formation and cell signaling. PUBLIC HEALTH RELEVANCE: Bacterial vaginosis affects 29% of women in the United States and is associated with increased risk of sexually transmitted diseases, including HIV, and increased risk of preterm birth, as well as other adverse outcomes. It is unclear why some women develop BV and why affected women have a high rate of recurrence after antibiotic treatment. This proposal will use newly developed molecular tools to study the microbial and host factors that lead to BV, which will create new opportunities for prevention and treatment of this extremely common condition.