A Coxsackievirus B1-mediated nonlytic Extracellular Vesicle-to-cell mechanism of virus transmission and its possible control through modulation of EV release

Author:

Jorfi Samireh1,Ansa-Addo Ephraim Abrokwa21,Mariniello Katia31,Warde Purva4,bin Senian Ahmad Asyraf54,Stratton Dan6,Bax Bridget E.7ORCID,Levene Michelle7,Lange Sigrun89ORCID,Inal Jameel Malhador14ORCID

Affiliation:

1. Cell Communication in Disease Pathology, School of Human Sciences, London Metropolitan University, London N7 8DB, UK

2. Present address: Pelotonia Institute for Immuno-Oncology, The James, Ohio State University, Columbus, OH 43210, USA

3. Present address: William Harvey Research Institute, Queen Mary, University of London, London, UK

4. Biosciences Research Group, School of Life and Medical Sciences, University of Hertfordshire, Hatfield AL10 9EU, UK

5. Present address: Clinical Research Centre, Sarawak General Hospital, Kuching, Malaysia

6. School of Life, Health & Chemical Sciences, The Open University, Milton Keynes MK7 6AE, UK

7. Molecular and Clinical Sciences Research Institute, St. George’s, University of London, London SW17 0RE, UK

8. University College London School of Pharmacy, Brunswick Sq., London, UK

9. Tissue Architecture and Regeneration Research Group, School of Life Sciences, University of Westminster, 116, New Cavendish St., London, UK

Abstract

Like most non-enveloped viruses, CVB1 mainly uses cell lysis to spread. Details of a nonlytic virus transmission remain unclear. Extracellular Vesicles (EVs) transfer biomolecules between cells. We show that CVB1 entry into HeLa cells results in apoptosis and release of CVB1-induced ‘medium-sized’ EVs (CVB1i-mEVs). These mEVs (100–300 nm) harbour CVB1 as shown by immunoblotting with anti-CVB1-antibody; viral capsids were detected by transmission electron microscopy and RT-PCR revealed CVB1 RNA. The percentage of mEVs released from CVB1-infected HeLa cells harbouring virus was estimated from TEM at 34 %. Inhibition of CVB1i-mEV production, with calpeptin or siRNA knockdown of CAPNS1 in HeLa cells limited spread of CVB1 suggesting these vesicles disseminate CVB1 virions to new host cells by a nonlytic EV-to-cell mechanism. This was confirmed by detecting CVB1 virions inside HeLa cells after co-culture with CVB1i-mEVs; EV release may also prevent apoptosis of infected cells whilst spreading apoptosis to secondary sites of infection.

Funder

Research Executive Agency

Publisher

Microbiology Society

Subject

Virology

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