Velocity Gradient Separation Reveals a New Extracellular Vesicle Population Enriched in miR-155 and Mitochondrial DNA Article Swipe
Myriam Vaillancourt
,
Audrey Hubert
,
Caroline Subra
,
Julien Boucher
,
Wilfried Wenceslas Bazié
,
Julien Vitry
,
Sofiane Berrazouane
,
Jean‐Pierre Routy
,
Sylvie Trottier
,
Cécile Tremblay
,
Mohammad‐Ali Jenabian
,
Abderrahim Benmoussa
,
Patrick Provost
,
Philippe A. Tessier
,
Caroline Gilbert
·
YOU?
·
· 2021
· Open Access
·
· DOI: https://doi.org/10.3390/pathogens10050526
YOU?
·
· 2021
· Open Access
·
· DOI: https://doi.org/10.3390/pathogens10050526
Extracellular vesicles (EVs) and their contents (proteins, lipids, messenger RNA, microRNA, and DNA) are viewed as intercellular signals, cell-transforming agents, and shelters for viruses that allow both diagnostic and therapeutic interventions. EVs circulating in the blood of individuals infected with human immunodeficiency virus (HIV-1) may provide insights into pathogenesis, inflammation, and disease progression. However, distinguishing plasma membrane EVs from exosomes, exomeres, apoptotic bodies, virions, and contaminating proteins remains challenging. We aimed at comparing sucrose and iodixanol density and velocity gradients along with commercial kits as a means of separating EVs from HIV particles and contaminating protein like calprotectin; and thereby evaluating the suitability of current plasma EVs analysis techniques for identifying new biomarkers of HIV-1 immune activation. Multiple analysis have been performed on HIV-1 infected cell lines, plasma from HIV-1 patients, or plasma from HIV-negative individuals spiked with HIV-1. Commercial kits, the differential centrifugation and density or velocity gradients to precipitate and separate HIV, EVs, and proteins such as calprotectin, have been used. EVs, virions, and contaminating proteins were characterized using Western blot, ELISA, RT-PCR, hydrodynamic size measurement, and enzymatic assay. Conversely to iodixanol density or velocity gradient, protein and virions co-sedimented in the same fractions of the sucrose density gradient than AChE-positive EVs. Iodixanol velocity gradient provided the optimal separation of EVs from viruses and free proteins in culture supernatants and plasma samples from a person living with HIV (PLWH) or a control and revealed a new population of large EVs enriched in microRNA miR-155 and mitochondrial DNA. Although EVs and their contents provide helpful information about several key events in HIV-1 pathogenesis, their purification and extensive characterization by velocity gradient must be investigated thoroughly before further use as biomarkers. By revealing a new population of EVs enriched in miR-155 and mitochondrial DNA, this study paves a way to increase our understanding of HIV-1 pathogenesis.
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Metadata
- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.3390/pathogens10050526
- https://www.mdpi.com/2076-0817/10/5/526/pdf?version=1619522303
- OA Status
- gold
- Cited By
- 14
- References
- 68
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W3158910500
All OpenAlex metadata
Raw OpenAlex JSON
- OpenAlex ID
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https://openalex.org/W3158910500Canonical identifier for this work in OpenAlex
- DOI
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https://doi.org/10.3390/pathogens10050526Digital Object Identifier
- Title
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Velocity Gradient Separation Reveals a New Extracellular Vesicle Population Enriched in miR-155 and Mitochondrial DNAWork title
- Type
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articleOpenAlex work type
- Language
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enPrimary language
- Publication year
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2021Year of publication
- Publication date
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2021-04-27Full publication date if available
- Authors
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Myriam Vaillancourt, Audrey Hubert, Caroline Subra, Julien Boucher, Wilfried Wenceslas Bazié, Julien Vitry, Sofiane Berrazouane, Jean‐Pierre Routy, Sylvie Trottier, Cécile Tremblay, Mohammad‐Ali Jenabian, Abderrahim Benmoussa, Patrick Provost, Philippe A. Tessier, Caroline GilbertList of authors in order
- Landing page
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https://doi.org/10.3390/pathogens10050526Publisher landing page
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https://www.mdpi.com/2076-0817/10/5/526/pdf?version=1619522303Direct link to full text PDF
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YesWhether a free full text is available
- OA status
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goldOpen access status per OpenAlex
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https://www.mdpi.com/2076-0817/10/5/526/pdf?version=1619522303Direct OA link when available
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Microvesicles, Population, Western blot, Biology, Density gradient, Differential centrifugation, Extracellular, Cell biology, Molecular biology, Chemistry, Virology, microRNA, Biochemistry, Gene, Medicine, Physics, Quantum mechanics, Environmental healthTop concepts (fields/topics) attached by OpenAlex
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14Total citation count in OpenAlex
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2025: 6, 2024: 2, 2023: 3, 2022: 3Per-year citation counts (last 5 years)
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68Number of works referenced by this work
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10Other works algorithmically related by OpenAlex
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