Mechanisms of Sexual HIV-1 Transmission

个人描述(申请人提供)：自从我开始关心艾滋病病毒-L 1993年感染的患者，令我印象深刻的是尽管 粘膜传播HIV-1的相对低效，这主要是 异性传播的全球疫情继续有增无减 有效的疫苗或杀微生物剂。设计中的一个基本障碍 预防策略是我们缺乏关于传播的知识 整个人类生殖器粘膜的事件。此K08应用程序将支持我的 努力阐明L艾滋病病毒的性传播机制 加强我在分子免疫学和病毒学方面的培训。我感觉到了这一点 培训对于实现我的长期研究目标是必不可少的，那就是 翻译人类泌尿生殖免疫系统的许多方面的知识 及其与艾滋病病毒的互动-L成为中断艾滋病病毒-L的战略 变速箱。 树突状细胞(DC)和CD4+T细胞被认为与 HIV-I在性传播过程中的重要目标，但直接证据 因为这在人类生殖器粘膜中是缺乏的。此外，这些机制 病毒株的选择，特别是病毒的优先传播 R5嗜性HIV-I变异体，人们对此知之甚少。我们是第一个 从人生殖器粘膜中成功分离DC和T细胞 但我们对HIV-L共受体表达和体外感染性的研究 没有解释传播过程中的病毒选择。潜在的陷阱是 在长达36小时的细胞分离所需的体外迁移程序中， 这可能导致了 人类免疫缺陷病毒-L共受体表达的变化及无法区分 来自基质细胞的上皮内。为了绕过这些限制，我们有 建立了从阴道、外周和外周分离单个核细胞的新程序 术后3小时内行宫颈腔内注射，并清楚地勾画出 来源于上皮和下层基质的细胞。在特定的目标中 1和2的应用，我们将研究表型和功能 T细胞和树突状细胞在人体粘膜中的浸润特性 生殖道。在目标1中，我们将比较归巢受体的配置文件， 免疫分化和激活状态，以及抗原特异性 阴道上皮、间质、外膜和间质中的T细胞 子宫内膜，以及循环中的T细胞。在目标2中，我们将定义阶段 DC成熟和描绘DC谱系和分化途径的研究 人类生殖器粘膜。在目标3中，我们将讲述HIV-L的表达谱 新鲜分离的人上皮内和间质白细胞上的辅受体 生殖器粘膜具有传染性，具有R5和X4嗜性的艾滋病毒-L变异株。这些 研究将提高我们对人类生殖器免疫系统的了解 HIV-L在女性体内的靶向和毒株选择机制 生殖道。

DESCRIPTION (provided by applicant): Since I first started caring for HIV-l infected patients in 1993, I am impressed by the fact that despite the relative inefficiency of mucosal HIV-1 transmission, this mostly heterosexually transmitted global epidemic continues unabated without an effective vaccine or microbicide. One fundamental hurdle in designing prevention strategies is our lack of knowledge concerning the transmission events across the human genital mucosa. This K08 application will support my endeavor to elucidate the mechanisms of sexual HIV-l transmission while enhancing my training in molecular immunology and virology. I perceive this training as essential to accomplish my long-term research goal, which is to translate knowledge of the many facets of the human urogenital immune system and its interaction with HIV-l into strategies to interrupt HIV-l transmission. Dendritic cells (DC) and CD4+ T cells have been strongly implicated as important targets for HIV-I during sexual transmission, but direct evidence for this in the human genital mucosa is lacking. In addition, the mechanisms of viral strain selection, and particularly the preferential transmission of R5-tropic HIV-I variants, are poorly understood. We were the first to successfully isolate DC and T cells from the human genital mucosa but our studies on HIV-l coreceptor expression and in vitro infectivity did not explain viral selection during transmission. Potential pitfalls were the 36-hour long in vitroemigration procedure required for cell isolation, which may have resulted in changes of HIV-l coreceptor expression, and the inability to distinguish intraepithelial from stromal cells. To circumvent these limitations, we have established new procedures to isolate mononuclear cells from the vagina, ecto-and endocervix within three hours after surgery, and to clearly delineate cells derived from the epithelium and the underlying stroma. In Specific Aims 1 and 2 of this application, we will investigate the phenotypic and functional properties of T cells and DC infiltrating the mucosa of the human female genital tract. In Aim 1, we will compare the homing receptor profile, the state of immune differentiation and activation, and the antigen-specific repertoire of T cells among the epithelium and stroma of vagina, ecto- and endocervix, and to T cells in circulation. In Aim 2, we will define the stages of DC maturation and delineate DC lineage and differentiation pathways in the human genital mucosa. In Aim 3, we will relate the expression profile of HIV-l coreceptors on intraepithelial and stromal leukocytes isolated freshly from the genital mucosa to infectivity with R5- and X4-tropic HIV-l variants. These studies will improve our knowledge of the human genital immune system and shed light on the mechanisms of HIV-l targeting and strain selection in the female genital tract.