A well characterized simian immunodeficiency virus (SIV)/macaque model of HIV was used to profile extracellular vesicle enriched blood plasma fractions harvested during preinfection, acute infection, latent infection/ART treatment, and rebound after ART interruption.
Measurement of extracellular vesicle concentration, size distribution, and morphology was complemented with qPCR array for small RNA expression, followed by individual qPCR validations. Iodixanol density gradients were used to separate extracellular vesicle subtypes and virions.
Plasma extracellular vesicle particle counts correlated with viral load and peaked during acute infection. However, SIV gag RNA detection showed that virions did not fully explain this peak. Extracellular vesicle microRNAs miR-181a, miR-342-3p, and miR-29a decreased with SIV infection and remained downregulated in latency. Interestingly, small nuclear RNA U6 had a tight association with viral load peak.
This study is the first to monitor how extracellular vesicle concentration and extracellular vesicle small RNA expression change dynamically in acute viral infection, latency, and rebound in a carefully controlled animal model. These changes may also reveal regulatory roles in retroviral infection and latency.
方法:使用特征良好的猿猴免疫缺陷病毒(SIV)/猕猴模型对感染前收获的富含EV的血浆部分进行分析,急性感染,潜伏感染/ART治疗,ART中断后反弹。
方法:EV浓度的测量,大小分布,和形态学用qPCR阵列补充小RNA表达,其次是单独的qPCR验证。碘克沙醇密度梯度用于分离EV亚型和病毒体。
结果:血浆EV颗粒计数与病毒载量相关,并在急性感染期间达到峰值。然而,SIVgagRNA检测显示病毒体不能完全解释该峰。EVmicroRNAsmiR-181a,miR-342-3p,miR-29a随SIV感染而降低,并在潜伏期保持下调。有趣的是,小核RNAU6与病毒载量峰值密切相关.
结论:本研究首次监测急性病毒感染中EV浓度和EV小RNA表达的动态变化,延迟,在精心控制的动物模型中反弹。这些变化也可能揭示在逆转录病毒感染和潜伏期中的调节作用。