Cataloguing of Potential HIV Susceptibility Factors during the Menstrual Cycle of Pigtail Macaques. Macaca nemestrina

Our earlier studies in pig-tailed macaques demonstrated varying SHIV susceptibility during the menstrual cycle, likely caused by cyclic variations in immune responses in the female genital tract. There is concern that high-dose, long-lasting, injectable progestin-based contraception could mimic the luteal phase and predispose women to HIV-1 acquisition and transmission. In this study, we adopted a systems biology approach employing proteomics (tandem mass spectrometry), transcriptomics (RNA microarray hybridization), and other specific protein assays (enzyme-linked immunosorbent assays and multiplex chemokine-cytokine measurements) to characterize the effects of hormonal changes on the expression of innate factors and secreted proteins in the macaque vagina. Several antiviral factors and pathways (including acute phase response signaling and complement system) were overexpressed in the follicular phase. Conversely, during the luteal phase there were factors overexpressed (including moesins, syndecans, integrins, among others) that could play direct or indirect roles in enhancing HIV-1 infection. Thus, our study showed that specific pathways and proteins/genes might be working in tandem to regulate innate immunity, thus fostering further investigation and future design of approaches to help counter HIV-1 acquisition in the female genital tract. Overall design: Samples were hybridized to Affymetrix GeneChip® Rhesus Macaque Genome Arrays. Vaginal pinch biopsies were collected from 12 pig-tailed macaques at both the follicular and luteal phases. The data from one animal suggested low RNA quality and was excluded.

我们此前在豚尾猴（pig-tailed macaque）中的研究表明，其在月经周期内对猴-人免疫缺陷病毒（SHIV）的易感性存在差异，该差异大概率由雌性生殖道免疫应答的周期性变化所引发。
有研究担忧，高剂量长效注射型孕激素类避孕药可能模拟黄体期状态，从而增加女性感染并传播1型人类免疫缺陷病毒（HIV-1）的风险。
本研究采用系统生物学（systems biology）方法，结合采用串联质谱（tandem mass spectrometry）技术的蛋白质组学（proteomics）、采用RNA微阵列杂交（RNA microarray hybridization）技术的转录组学（transcriptomics），以及酶联免疫吸附试验（enzyme-linked immunosorbent assays）、多重趋化因子-细胞因子检测（multiplex chemokine-cytokine measurements）等特异性蛋白质检测技术，解析激素变化对豚尾猴阴道内先天因子与分泌蛋白表达的影响。
在卵泡期，多种抗病毒因子及信号通路（包括急性期应答信号通路与补体系统）呈现过表达状态。
与之相反，黄体期内多种因子（包括膜突蛋白（moesins）、黏结蛋白聚糖（syndecans）、整合素（integrins）等）呈现过表达，这些因子可通过直接或间接途径促进HIV-1感染。
综上，本研究表明特定的信号通路与蛋白质/基因可能协同调控先天免疫，这为后续开展相关研究以及设计抵御女性生殖道HIV-1感染的干预策略提供了依据。
实验整体设计：样本与Affymetrix GeneChip®恒河猴基因组芯片进行杂交。
本研究从12只豚尾猴的卵泡期与黄体期分别采集阴道钳夹活检标本。
其中1只实验动物的样本因RNA质量过低被排除。