IN VITRO MODEL OF HUMAN VAGINAL EPITHELIUM

描述(改编自申请者的描述)：人的阴道 上皮位于一个关键的解剖位置，在外部 在上生殖道的环境和受保护的地点，并形成一个 防止感染病原体的屏障。一个健康的阴道生态系统对 生殖健康。雌激素替代疗法和多种避孕药 产品和设备是经阴道交付的，可能会影响阴道 上皮组织。尽管阴道上皮在生殖系统中的重要性 健康与疾病、阴道生理及影响分化的因素 对阴道上皮的研究很少，而且体外模型 这种组织还没有得到广泛的开发。申请人已被卷入其中 自1995年在HD33202中，一项临床研究定义了未探索的参数 正常的阴道生理，定义了由普通阴道产品引起的变化， 并调查了这些产品对性易感性的影响 传播疾病。作为这项研究的结果，申请者已经 开发了一种原代培养的阴道细胞(VECs)模型并进行了 细菌黏附及外源性雌激素对其影响的初步研究 细胞的生长和分化。她的初步数据显示， 培养的血管内皮细胞对细菌黏附的敏感性因(A)而不同 分化的形态参数；(B)表达 神经鞘糖脂(GSLS)，作为分化标志物和 细菌结合部位；及(C)外源性雌激素的剂量和时间 曝光。这一小笔赠款提案的目标是开发这一前景看好的 上皮分化的基础研究模型和 对细菌感染的敏感性，在体外暴露于 荷尔蒙和阴道产品。申请人假设这种区别 阴道上皮的变化影响其对感染的敏感性和反应 外源激素和避孕产品的应用。这个 申请者将追求以下目标：(1)初级血管内皮细胞将在 并将进一步特色化，以表达差异化 标志物，如角蛋白，上皮分化标志物广泛存在 在相关的上皮细胞中进行了研究，并选择了已知参与的GSLs 细菌粘连。正常年轻女性的阴道组织切片， 收集作为HD33202的一部分，将被染色为相同的特征； (2)外源性雌激素应用于体外培养的血管内皮细胞及参数 上皮分化，如角蛋白和GSLS的表达， 将被调查；(3)乳杆菌和沙门氏菌黏附的研究 细菌病原体，如大肠杆菌，可能与早产风险有关 出生，将扩大到包括表达之间的关系 分化标志物与外源性雌激素对易感性的影响 细菌附着物。建立这一模式将为 研究妇女泌尿生殖健康的多个方面，包括检测 阴道产品、益生菌和避孕药.了解防护 生物体在正常菌群中的作用；以及研究 荷尔蒙替代疗法的关键目标组织是阴道。

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.