IN VITRO MODEL OF HUMAN VAGINAL EPITHELIUM

描述（改编自申请人的描述）：人类阴道 上皮位于关键的解剖位置，在外部 在上生殖道的环境和受保护的网站，并形成一个 感染病原体的屏障。健康的阴道生态系统对 生殖健康。雌激素替代疗法和许多避孕药 产品和器械是经阴道输送的， 上皮尽管阴道上皮在生殖过程中的重要性 健康和疾病，阴道生理学和影响分化的因素 阴道上皮细胞的研究很少， 这种组织还没有被广泛开发。申请人曾参与 自1995年以来，在HD 33202中，一项临床研究定义了 正常的阴道生理学，定义了由普通阴道产品引起的变化， 并调查这些产品对性敏感性的影响， 传播疾病作为这项研究的结果，申请人 开发了原代培养阴道细胞（VEC）模型，并进行了 细菌粘附的初步研究和外源性雌激素对 细胞生长和分化。她的初步数据显示， 培养的VEC对细菌粘附的敏感性随（a） 分化的形态学参数;（B） 鞘糖脂（GSL），其用作分化标志物， 细菌结合位点;和（c）外源性雌激素的剂量和时间 exposure.这项小额赠款提案的目标是发展这种有前途的 上皮分化的基础研究模型， 在体外暴露于 激素和阴道产品。申请人假设， 阴道上皮细胞对感染的敏感性及其反应 外源激素和避孕产品的应用。的 申请人将追求以下目标：（1）初级VEC将在 并将进一步表征分化的表达 标记物，如角蛋白，上皮分化标记物，广泛地 在相关上皮中进行了研究，并选择了已知参与 细菌粘附。正常年轻女性的阴道组织切片 作为HD 33202的一部分采集，将针对相同特征进行染色; (2)将外源性雌激素应用于培养中生长的VEC， 上皮分化，如角蛋白和GSL的表达， （3）乳酸杆菌粘附性研究， 细菌病原体如E.大肠杆菌，这可能与早产的风险有关。 出生，将扩大到包括表达之间的关系， 分化标志物和外源性雌激素对易感性的影响 到细菌附着。建立这种模式将为以下方面提供机会： 研究女性泌尿生殖健康的许多方面，包括测试 阴道产品，益生菌和避孕药;了解保护 生物体在正常植物群中的作用;以及研究 激素替代疗法的一个关键目标组织，阴道。

DESCRIPTION (Adapted from applicant's description): The human vaginal epithelium is located in a key anatomical location, between the external environment and protected sites in the upper genital tract, and forms a barrier to infecting pathogens. A healthy vaginal ecosystem is critical to reproductive health. Estrogen replacement therapy and many contraceptive products and devices are vaginally delivered and may affect the vaginal epithelium. Despite the importance of the vaginal epithelium in reproductive health and disease, vaginal physiology and factors affecting differentiation of the vaginal epithelium are infrequently studied, and in vitro models of this tissue have not been widely developed. The applicant has been involved since 1995 in HD33202, a clinical study that defines unexplored parameters of normal vaginal physiology, defines changes induced by common vaginal products, and investigates these products effects on susceptibility to sexually transmitted diseases. As an outgrowth of this study, the applicant has developed a model of primary cultured vaginal cells (VECs) and has performed pilot studies of bacterial adherence and the effects of exogenous estrogen on cell growth and differentiation. Her preliminary data indicate that susceptibility of cultured VEC to bacterial adherence varies with (a) morphological parameters of differentiation; (b) the expression of glycosphingolipids (GSLs), which serve as differentiation markers and bacterial binding sites; and (c) dose and timing of exogenous estrogen exposure. The goal of this Small Grant proposal is to develop this promising model for basic investigations of epithelial differentiation and susceptibility to bacterial infection, before and after in vitro exposure to hormonal and vaginal products. The applicant hypothesizes that differentiation of vaginal epithelium affects its susceptibility to infection and its response to the application of exogenous hormones and contraceptive products. The applicant will pursue the following aims: (1) Primary VECs will be grown in culture and will be further characterized for expression of differentiation markers, such as keratins, epithelial differentiation markers extensively studied in related epithelia, and selected GSLs known to be involved in bacterial adherence. Vaginal tissue sections from normal young women, collected as part of HD33202, will be stained for the same characteristics; (2) Exogenous estrogen will be applied to VECs grown in culture and parameters of epithelial differentiation, such as the expression of keratins and of GSLs, will be investigated; (3) Studies of the adherence of Lactobacilli and of bacterial pathogens such as E. coli, which may relate to risk of premature birth, will be expanded to include the relationship between expression of differentiation markers and the effect of exogenous estrogen on susceptibility to bacterial attachment. Establishing this model will open opportunities for studying numerous aspects of women's urogenital health, including testing vaginal products, probiotics, and contraceptives; understanding protective roles of organisms in the normal flora; and studying the cellular effects of hormone replacement therapy on a key target tissue, the vagina.