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J Virol. 2010 Nov;84(21):10999-1009. doi: 10.1128/JVI.00526-10. Epub 2010 Aug 4.

Ultrastructural and biophysical characterization of hepatitis C virus particles produced in cell culture.

Author information

1
Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, California 92037, USA. pgastaminza@cnb.csic.es

Abstract

We analyzed the biochemical and ultrastructural properties of hepatitis C virus (HCV) particles produced in cell culture. Negative-stain electron microscopy revealed that the particles were spherical (∼40- to 75-nm diameter) and pleomorphic and that some of them contain HCV E2 protein and apolipoprotein E on their surfaces. Electron cryomicroscopy revealed two major particle populations of ∼60 and ∼45 nm in diameter. The ∼60-nm particles were characterized by a membrane bilayer (presumably an envelope) that is spatially separated from an internal structure (presumably a capsid), and they were enriched in fractions that displayed a high infectivity-to-HCV RNA ratio. The ∼45-nm particles lacked a membrane bilayer and displayed a higher buoyant density and a lower infectivity-to-HCV RNA ratio. We also observed a minor population of very-low-density, >100-nm-diameter vesicular particles that resemble exosomes. This study provides low-resolution ultrastructural information of particle populations displaying differential biophysical properties and specific infectivity. Correlative analysis of the abundance of the different particle populations with infectivity, HCV RNA, and viral antigens suggests that infectious particles are likely to be present in the large ∼60-nm HCV particle populations displaying a visible bilayer. Our study constitutes an initial approach toward understanding the structural characteristics of infectious HCV particles.

PMID:
20686033
PMCID:
PMC2953183
DOI:
10.1128/JVI.00526-10
[Indexed for MEDLINE]
Free PMC Article

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