VAGINAL TRANSMISSION OF A PATHOGENIC RT-SHIVTC IN CYNOMOLGUS MACAQUES

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 中心，不一定是研究者的机构。 本项目的目的是开发一种经验证的猕猴模型，用于通过阴道内（ivag）和直肠内（ir）途径使用含有HIV-1 RT的充分表征的RT-SHIV进行HIV的性传播。本项目的基本原理和意义在"背景和意义"一节中说明。考虑到预算限制，本项目将侧重于食蟹猴（Macaca fascicularis）中RT-SHIV的ivag传播。我们将确定ivag暴露后病毒对全身感染性的剂量反应。感染的主要参数将包括外周血和外宫颈、阴道、直肠和肠道粘膜中的病毒RNA、病毒DNA和感染细胞。我们将表征外周血和阴道、宫颈外口和直肠（通过定期灌洗）以及小肠（尸检时）和周围淋巴结的粘膜表面中RT-SHIV的全身和局部感染的性质。感染的这些次要参数是为了鉴定不同粘膜部位（外宫颈、阴道、直肠和小肠（空肠和回肠））的感染细胞群和辅助因子（即促进病毒感染的细胞因子和趋化因子）。感染的其他次要参数是比较粘膜和全身对RT-SHIV的免疫应答;因此我们将测量阴道和直肠分泌物中以及从外周血获得的血清中SIV抗体的滴度，并检测粘膜和/或血清中产生的病毒蛋白的特异性抗体。总之，本研究的目的是最终通过阴道传播在食蟹猴中建立成功的RT-SHIV感染模型。此外，这种建立的RT-SHIV猕猴模型不仅可用于基于NNRTI的杀微生物剂候选物的疗效评价，而且可用于NNRTI和NRTI的全身化疗。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The objective of this project is to develop a validated macaque model for sexual transmission of HIV by intravaginal (ivag) and intrarectal (ir) routes using a well-characterized RT-SHIV containing the RT of HIV-1. The rationale and significance of this project are described in the "Background and Significance" section. Considering the budget restriction, this project will focus on an ivag transmission of RT-SHIV in cynomolgus macaques (Macaca fascicularis). We will determine the virus-dose response to systemic infectivity after ivag exposure. The primary parameters of infection will include viral RNA, viral DNA and infectious cells in peripheral blood and the mucosa of the ectocervix, vagina, rectum and gut. We will characterize the nature of systemic and local infection of RT-SHIV in peripheral blood and mucosal surfaces of vagina, ectocervix, and rectum (by periodic lavage) and small intestines (at necropsy) and surrounding lymph nodes. These secondary parameters of infection are to identify the infected cell population and accessory factors (i.e. cytokines and chemokines which facilitate virus infection) at different mucosal sites: ectocervix, vagina, rectum and small intestine (jejunum and ileum). Additional secondary parameters of infection are to compare mucosal and systemic immune responses to RT- SHIV; so we will measure titers of SIV-antibodies in vaginal and rectal secretions and in serum obtained from peripheral blood, and detect specific antibodies to viral proteins produced in mucosa and/or serum. In summary, the objective of this study is ultimately to develop the successful RT-SHIV infection model in the cynomolgus macaque by vaginal transmission. Moreover, this established RT-SHIV macaque model will be useful for the efficacy evaluations of not only NNRTI-based microbicide candidates but also NNRTI and NRTI for systemic chemotherapy.