E-105 Naked virions, extracellular vesicles, and vesicle-enclosed virions released early after picornavirus infection—who, when, how, and why?

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There is rising interest in the role of Extracellular Vesicles (EV) in virus infections. Viruses affect EV release by infected cells by influencing the incorporation of proteins and RNA from both host and viral origin into EV. Moreover, recent evidence suggests that naked viruses can exit the cell enclosed in EV (EV-virus hybrids). Enclosure of virions and viral components in EV can largely impact antiviral immunity. Still, the release kinetics of EV-virus hybrids and virus-induced EV, as well as the viral and/or host factors that regulate their formation and release, remain elusive. We addressed these topics using mengovirus, a non-enveloped, lytic RNA virus of the Picornaviridae family. A major technical hurdle in EV-virus research is the difficulty to reliably separate naked virus, (virus-induced) EV, and EV-virus hybrids due to their overlapping sizes. Our laboratory specializes in techniques to isolate subpopulations of EV, and to characterize these by general protein or RNA determination or, at the single particle level, by an in-house developed high-resolution flow cytometry (hFC) method. Using these methodologies, we discovered that early in mengovirus infection, hours before lytic release of virus, infectious particles were released from intact cells. These early stage infectious particles could be separated into populations of naked virions and EV-virus hybrids. In addition, quantitative and qualitative particle analysis by hFC indicated that virus infection induced the release of several EV subpopulations not observed in mock-infected samples. In- depth characterization of virus-induced EV and EV-virus hybrids aids our understanding of how EV contribute to viral dissemination and antiviral responses.

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