Detection of HIV-1 and Human Proteins in Urinary Extracellular Vesicles from HIV+ Patients.

IF 1.1 Q4 VIROLOGY Advances in Virology Pub Date : 2018-03-12 eCollection Date: 2018-01-01 DOI:10.1155/2018/7863412

Samuel I Anyanwu, Akins Doherty, Michael D Powell, Chamberlain Obialo, Ming B Huang, Alexander Quarshie, Claudette Mitchell, Khalid Bashir, Gale W Newman

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引用次数: 21

Abstract

Background: Extracellular vesicles (EVs) are membrane bound, secreted by cells, and detected in bodily fluids, including urine, and contain proteins, RNA, and DNA. Our goal was to identify HIV and human proteins (HPs) in urinary EVs from HIV+ patients and compare them to HIV- samples.

Methods: Urine samples were collected from HIV+ (n = 35) and HIV- (n = 12) individuals. EVs were isolated by ultrafiltration and characterized using transmission electron microscopy, tandem mass spectrometry (LC/MS/MS), and nanoparticle tracking analysis (NTA). Western blots confirmed the presence of HIV proteins. Gene ontology (GO) analysis was performed using FunRich and HIV Human Interaction database (HHID).

Results: EVs from urine were 30-400 nm in size. More EVs were in HIV+ patients, P < 0.05, by NTA. HIV+ samples had 14,475 HPs using LC/MS/MS, while only 111 were in HIV-. HPs in the EVs were of exosomal origin. LC/MS/MS showed all HIV+ samples contained at least one HIV protein. GO analysis showed differences in proteins between HIV+ and HIV- samples and more than 50% of the published HPs in the HHID interacted with EV HIV proteins.

Conclusion: Differences in the proteomic profile of EVs from HIV+ versus HIV- samples were found. HIV and HPs in EVs could be used to detect infection and/or diagnose HIV disease syndromes.

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HIV+患者尿细胞外囊泡中HIV-1和人蛋白的检测

背景:细胞外囊泡(EVs)是膜结合的，由细胞分泌，可在体液(包括尿液)中检测到，含有蛋白质、RNA和DNA。我们的目的是鉴定HIV+患者尿ev中的HIV和人蛋白(hp)，并将其与HIV-样本进行比较。方法:收集HIV阳性(n = 35)和HIV阴性(n = 12)患者的尿液样本。采用超滤分离ev，并采用透射电镜、串联质谱(LC/MS/MS)和纳米颗粒跟踪分析(NTA)对ev进行表征。Western blots证实了HIV蛋白的存在。使用FunRich和HIV Human Interaction database (HHID)进行基因本体(GO)分析。结果:尿液中ev大小为30 ~ 400 nm。经NTA分析，HIV阳性患者中EVs较多，P < 0.05。LC/MS/MS检测HIV+样品中有14475个HPs，而HIV-样品中只有111个HPs。EVs中的hp来源于外泌体。LC/MS/MS显示所有HIV+样品至少含有一种HIV蛋白。GO分析显示HIV+和HIV-样品之间的蛋白质存在差异，并且HHID中超过50%的已发表的hp与EV HIV蛋白相互作用。结论:HIV+和HIV-样本中EVs的蛋白质组学特征存在差异。EVs中的HIV和HPs可用于检测感染和/或诊断HIV疾病综合征。

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