Gardnerella vaginalis small colony variants

项目摘要 细菌性阴道病（BV）是育龄妇女最常见的妇科疾病， 与不良妊娠结局和性传播疾病传播增加有关。 阴道加德纳菌是与BV相关的标志性细菌，但其感染机制 和持久性是不太了解的。此外，还探讨了G.迷走神经感染复发 抗生素治疗后的情况尚不清楚。我们发现G.迷走神经以两种不同的形式生长， 生长缓慢的小菌落变体（SCV）和生长较快的大菌落变体（LCV）。SCV形式 产生更多的毒素阴道溶素，更具抗生素抗性，并且对 乳杆菌属LCV产生更多的生物膜，对奈瑟菌的生长具有更强的抑制作用 淋病初步的蛋白质组学分析表明，SCV下调DNA复制和蛋白质表达， 合成并上调阴道溶素和其他推定的毒力因子。我们假设SCV是一个 LCV是细菌的一种形式，它启动了感染或在不利条件下持续存在，而LCV是一种细菌， 提供更快的增殖并建立支持多种BV相关病原体的生物膜。为这些 研究，我们将描述SCV感染能力的差异，并使突变体表征 与变异形式的毒性增加和代谢改变有关的关键因素。发病机制 研究将包括在器官培养中感染人类宫颈组织。我们将确定涉及的机制， 使用蛋白质组学和抗生素耐药性测量， 并且我们将识别刺激相位变化的因素。

PROJECT SUMMARY Bacterial vaginosis (BV) is the most common gynecological disorder in women of childbearing age and is associated with adverse pregnancy outcomes and enhanced transmission of sexually-transmitted diseases. Gardnerella vaginalis is the signature bacterial species associated with BV, but its mechanisms of infection and persistence are not well understood. Furthermore, the mechanisms by which G. vaginalis infection recurs after antibiotic treatment are not known. We have discovered that G. vaginalis grows in two different forms, as a slow-growing small colony variant (SCV) and as a faster-growing large colony variant (LCV). The SCV form produces more of the toxin vaginolysin, is more antibiotic resistant, and is more inhibitory to the growth of Lactobacillus spp. The LCV produces more biofilms and is more inhibitory to the growth of Neisseria gonorrhoeae. Preliminary proteomic analyses indicate that SCVs downregulate DNA replication and protein synthesis and upregulate vaginolysin and other putative virulence factors. We hypothesize that the SCV is a form of the bacteria primed for initiating infection or for persisting under adverse conditions, while the LCV gives faster multiplication and builds the biofilm that supports multiple BV-associated pathogens. For these studies, we will characterize differences in infection abilities of the SCVs and make mutants to characterize the key factors involved in the increased virulence and altered metabolism of the variant form. Pathogenesis studies will include infection of human cervical tissue in organ culture. We will identify mechanisms involved in persistence and survival in adverse conditions using proteomics and measurements of antibiotic resistance, and we will identify factors stimulating phase variation.