2026-07-21T19:38:18Zhttps://repisalud.isciii.es/rest/oai/requestoai:repisalud.isciii.es:20.500.12105/110022024-11-28T18:55:59Zcom_20.500.12105_2052com_20.500.12105_2051com_20.500.12105_15322col_20.500.12105_19609col_20.500.12105_16963
Repisalud
author
Martinez-Gonzalez, Elena
author
Martín-Carbonero, Luz
author
Brochado-Kith, Oscar
author
Gomez-Sanz, Alicia
author
Jimenez-Sousa, Maria Angeles
author
Martinez-Roman, Paula
author
Resino, Salvador
author
Briz, Veronica
author
Fernandez-Rodriguez, Amanda
funder
Instituto de Salud Carlos III
2020-09-14T07:20:14Z
2020-09-14T07:20:14Z
2020-07-07
Sci Rep . 2020 Jul 7;10(1):11140.
10.1038/s41598-020-67935-1
2045-2322
Scientific reports
32636456
http://hdl.handle.net/20.500.12105/11002
Hepatitis C virus (HCV) and human immunodeficiency virus (HIV) hijack the host exosomal machinery as an additional mechanism of infection and evasion of the immune system, modifying the small RNA (smRNA) cargo during infection. We characterized the surface epitopes of extracellular vesicles (EVs) from plasma HIV/HCV-coinfected patients and their smRNA cargo profile, by comparing different isolation procedures. Six EVs isolation procedures were compared: ultracentrifugation, and five different polyethylene glycol-based methods (commercial, combined with a column purification step and two custom); and two RNA commercial kits (phenol and non-phenol based) were used. High-throughput sequencing of smRNAs was performed. Exosomal surface epitopes were analyzed by the MACSPlex Exosome Kit. Four miRNAs displayed differences among protocols (hsa-miR-205-5p and hsa-let-7a/b/f-5p). The selection of RNA isolation kit impacted on the detection of miRNAs and other smRNAs, where the phenol-based RNA isolation kit performed acceptably. EVs surface was enriched with HLA-DR/DP/DQ, CD81, and CD8. There were three liver-specific miRNAs overexpressed (let-7a-5p, miR-21-5p and hsa-miR-122-5p), thus, EVs cargo might reflect liver disease evolution. Other smRNAs such as piwi-interacting RNAs were also detected for the first time. Custom polyethylene glycol precipitation-based methods combined with an RNA phenol-based kit yielded the higher number of smRNAs for EVs isolated from plasma HIV/HCV patients.
eng
Comparison of methods and characterization of small RNAs from plasma extracellular vesicles of HIV/HCV coinfected patients.
journal article
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ComparisonOfMethodsAnd_2020.pdf
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https://repisalud.isciii.es/bitstreams/5bc178ea-13ec-46fe-bdfa-7c3d6e66f32f/download
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ComparisonOfMethodsAnd_2020.pdf.txt