Elucidating the mechanism of progesterone-induced permissivity in the upper female reproductive tract

项目摘要/摘要： 在世界范围内，妇女首当其冲地受到艾滋病毒流行的影响，而预防艾滋病毒传播的有效手段 女性仍然是一个难以捉摸的目标。实现这一目标受阻于我们对早期 性传播期间女性生殖道(FRT)的事件。特别是，居民艾滋病毒是如何- 敏感和不敏感细胞影响病毒复制，并对艾滋病毒感染做出感知和反应 很好理解。尽管上面的FRT含有艾滋病毒敏感细胞，但暴露在精液成分中， 并在SIV传播模型中早期感染，但作为艾滋病毒进入的门户研究不足。在自行车比赛中 女性，FRT上段的子宫内膜对性激素雌激素的反应广泛重塑 还有黄体酮。孕酮和孕酮是孕酮的合成模拟物，与 人类和非人类灵长类动物对艾滋病毒感染的易感性增加。因此， 了解这些化合物是如何影响子宫内膜的，可能会对HIV是如何建立的有深入的了解 通过这种组织感染。在我们的初步研究中，孕酮降低了子宫内膜的屏障功能 上皮细胞，它可以促进艾滋病毒从管腔进入子宫的间质隔室 子宫内膜。间质内有密集的子宫内膜间质成纤维细胞。尽管这些 细胞对HIV感染是不允许的，在与CD4T细胞共培养时，它们将感染增加高达81- 收牌。相反，当没有黄体酮时，上皮屏障的完整性保持不变，完好层 子宫内膜上皮细胞通过在共培养的CD4T细胞中诱导抗病毒状态来限制感染。vbl.使用 一个由子宫内膜活检组织的细胞重组的体外系统，我们将检查分子 黄体酮导致漏尿的机制(特异性目标1)，增加对 间质成纤维细胞介导的感染(特异性目标2)和上皮细胞诱导的病毒限制(特异性目标 3)FRT上段。由于孕酮诱导的上皮通透性也可能发生在较低的FRT， 我们还将使用从宫颈外活检中分离出的细胞来测试这一模型。拟议的研究将 加深我们对HIV在首次RT中传播和限制的理解，并提供一个强大的体外模型 在艾滋病毒背景下检查性类固醇和组织驻留细胞之间复杂相互作用的系统 敏感度。该系统和我们建立的表型分析工具也适用于HIV的研究 在其他粘膜入口的传播和限制，如肠粘膜和男性包皮。

PROJECT SUMMARY / ABSTRACT: Worldwide, women bear the brunt of the HIV epidemic, and an effective means to prevent HIV transmission to women remains an elusive goal. Achieving this goal is hampered by our incomplete understanding of the early events in the female reproductive tract (FRT) during sexual transmission. In particular, how resident HIV- susceptible and non-susceptible cells affect viral replication, and sense and respond to HIV infection, are not well understood. Although the upper FRT harbors HIV-susceptible cells, is exposed to semen components, and is infected early in SIV transmission models, it is understudied as a portal of entry for HIV. In cycling women, the endometrium of the upper FRT is extensively remodeled in response to the sex steroids estrogen and progesterone. Progesterone and progestins, synthetic mimics of progesterone, are associated with increased susceptibility to HIV infection in humans and in non-human primate models. Therefore, understanding how these compounds affect the endometrium may yield insights into how HIV establishes infection through this tissue. In our preliminary studies, progesterone decreased barrier function of endometrial epithelial cells, which can facilitate entry of HIV from the luminal cavity into the stromal compartment of the endometrium. Within the stroma resides a dense population of endometrial stromal fibroblasts. Although these cells were not permissive to HIV infection, in co-culture with CD4+ T cells they increased infection by up to 81- fold. In contrast, when progesterone was absent, epithelial barrier integrity was maintained, and the intact layer of endometrial epithelial cells restricted infection by eliciting an antiviral state in co-cultured CD4+ T cells. Using an ex vivo system reconstituted from cells from endometrial biopsies, we will examine the molecular mechanisms responsible for progesterone-induced leakiness (Specific Aim 1), the increased susceptibility to infection mediated by stromal fibroblasts (Specific Aim 2), and epithelia-induced viral restriction (Specific Aim 3) in the upper FRT. Because progesterone-induced epithelial permeability may also occur in the lower FRT, we will additionally test this model using cells isolated from ectocervical biopsies. The proposed studies will deepen our understanding of HIV transmission and restriction in the FRT and provide a robust ex vivo model system to examine the complex interplay between sex steroids and tissue-resident cells in the context of HIV susceptibility. This system and the phenotyping tools we have established can also be adapted to study HIV transmission and restriction in other mucosal portals of entry such the intestinal mucosa and the male foreskin.