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Complexity and ultrastructure of infectious extracellular vesicles from cells infected by non-enveloped virus.

Scientific reports (2020-05-16)
Jie E Yang, Evan D Rossignol, Deborah Chang, Joseph Zaia, Isaac Forrester, Kiran Raja, Holly Winbigler, Daniela Nicastro, William T Jackson, Esther Bullitt
ABSTRACT

Enteroviruses support cell-to-cell viral transmission prior to their canonical lytic spread of virus. Poliovirus (PV), a prototype for human pathogenic positive-sense RNA enteroviruses, and picornaviruses in general, transport multiple virions en bloc via infectious extracellular vesicles, 100~1000 nm in diameter, secreted from host cells. Using biochemical and biophysical methods we identify multiple components in secreted microvesicles, including mature PV virions; positive-sense genomic and negative-sense replicative, template viral RNA; essential viral replication proteins; and cellular proteins. Using cryo-electron tomography, we visualize the near-native three-dimensional architecture of secreted infectious microvesicles containing both virions and a unique morphological component that we describe as a mat-like structure. While the composition of these mat-like structures is not yet known, based on our biochemical data they are expected to be comprised of unencapsidated RNA and proteins. In addition to infectious microvesicles, CD9-positive exosomes released from PV-infected cells are also infectious and transport virions. Thus, our data show that, prior to cell lysis, non-enveloped viruses are secreted within infectious vesicles that also transport viral unencapsidated RNAs, viral and host proteins. Understanding the structure and function of these infectious particles helps elucidate the mechanism by which extracellular vesicles contribute to the spread of non-enveloped virus infection.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
2,2,2-Trifluoroethanol, ReagentPlus®, ≥99%
Sigma-Aldrich
Protease Inhibitor Cocktail, for use in tissue culture media, DMSO solution