Cervicovaginal secretions, or their components collected, are referred to as cervicovaginal lavage (CVL). CVL constituents have utility as biomarkers and play protective roles in wound healing and against HIV‐1 infection. However, several components of cervicovaginal fluids are less well understood, such as extracellular RNAs and their carriers, for example, extracellular vesicles (EVs). EVs comprise a wide array of double‐leaflet membrane extracellular particles and range in diameter from 30 nm to over one micron. The aim of this study was to determine whether differentially regulated CVL microRNAs (miRNAs) might influence retrovirus replication. To this end, we characterized EVs and miRNAs of primate CVL during the menstrual cycle and simian immunodeficiency virus (SIV) infection of macaques. EVs were enriched by stepped ultracentrifugation, and miRNA profiles were assessed with a medium‐throughput stem‐loop/hydrolysis probe qPCR platform. Whereas hormone cycling was abnormal in infected subjects, EV concentration correlated with progesterone concentration in uninfected subjects. miRNAs were present predominantly in the EV‐depleted CVL supernatant. Only a small number of CVL miRNAs changed during the menstrual cycle or SIV infection, for example, miR‐186‐5p, which was depleted in retroviral infection. This miRNA inhibited HIV replication in infected macrophages in vitro. In silico target prediction and pathway enrichment analyses shed light on the probable functions of miR‐186‐5p in hindering HIV infections via immunoregulation, T‐cell regulation, disruption of viral pathways, etc. These results provide further evidence for the potential of EVs and small RNAs as biomarkers or effectors of disease processes in the reproductive tract.

中文翻译：

---

灵长类动物宫颈阴道灌洗液和细胞外囊泡的 miRNA 分析揭示 miR-186-5p 作为巨噬细胞中潜在的抗逆转录病毒因子

宫颈阴道分泌物或其收集的成分被称为宫颈阴道灌洗液 (CVL)。CVL 成分可用作生物标志物，并在伤口愈合和抵抗 HIV-1 感染中发挥保护作用。然而，人们对宫颈阴道液的几种成分还不太了解，例如细胞外 RNA 及其载体，例如细胞外囊泡 (EV)。EV 包含多种双叶膜细胞外颗粒，直径范围从 30 nm 到超过 1 微米。本研究的目的是确定差异调节的 CVL microRNA (miRNA) 是否可能影响逆转录病毒复制。为此，我们对灵长类CVL在月经周期和猴免疫缺陷病毒（SIV）感染期间的EV和miRNA进行了表征。通过逐步超速离心富集 EV，并使用中等通量的茎环/水解探针 qPCR 平台评估 miRNA 图谱。尽管感染者的激素循环异常，但未感染者的 EV 浓度与孕酮浓度相关。miRNA 主要存在于 EV 耗尽的 CVL 上清液中。只有少量 CVL miRNA 在月经周期或 SIV 感染期间发生变化，例如 miR-186-5p，它在逆转录病毒感染中被耗尽。这种 miRNA在体外抑制受感染巨噬细胞中的 HIV 复制。计算机靶点预测和通路富集分析揭示了 miR-186-5p 通过免疫调节、T 细胞调节、病毒通路破坏等阻碍 HIV 感染的可能功能。这些结果为 EV 和病毒通路的潜力提供了进一步的证据。小RNA作为生殖道疾病过程的生物标志物或效应物。