Functional and Biochemical Characterization of Hepatitis C Virus (HCV) Particles Produced in a Humanized Liver Mouse Model

J Biol Chem. 2015 Sep 18;290(38):23173-87. doi: 10.1074/jbc.M115.662999. Epub 2015 Jul 29.

Abstract

Lipoprotein components are crucial factors for hepatitis C virus (HCV) assembly and entry. As hepatoma cells producing cell culture-derived HCV (HCVcc) particles are impaired in some aspects of lipoprotein metabolism, it is of upmost interest to biochemically and functionally characterize the in vivo produced viral particles, particularly regarding how lipoprotein components modulate HCV entry by lipid transfer receptors such as scavenger receptor BI (SR-BI). Sera from HCVcc-infected liver humanized FRG mice were separated by density gradients. Viral subpopulations, termed HCVfrg particles, were characterized for their physical properties, apolipoprotein association, and infectivity. We demonstrate that, in contrast to the widely spread distribution of apolipoproteins across the different HCVcc subpopulations, the most infectious HCVfrg particles are highly enriched in apoE, suggesting that such apolipoprotein enrichment plays a role for entry of in vivo derived infectious particles likely via usage of apolipoprotein receptors. Consistent with this salient feature, we further reveal previously undefined functionalities of SR-BI in promoting entry of in vivo produced HCV. First, unlike HCVcc, SR-BI is a particularly limiting factor for entry of HCVfrg subpopulations of very low density. Second, HCVfrg entry involves SR-BI lipid transfer activity but not its capacity to bind to the viral glycoprotein E2. In conclusion, we demonstrate that composition and biophysical properties of the different subpopulations of in vivo produced HCVfrg particles modulate their levels of infectivity and receptor usage, hereby featuring divergences with in vitro produced HCVcc particles and highlighting the powerfulness of this in vivo model for the functional study of the interplay between HCV and liver components.

Keywords: animal model; apolipoprotein; hepatitis C virus (HCV); in vivo study; lipoprotein; scavenger receptor; virus entry.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apolipoproteins E / genetics
  • Apolipoproteins E / metabolism
  • Cell Line
  • Disease Models, Animal
  • Hepacivirus / genetics
  • Hepacivirus / metabolism*
  • Hepatitis C / genetics
  • Hepatitis C / metabolism*
  • Hepatitis C / pathology
  • Humans
  • Liver / metabolism
  • Liver / pathology
  • Liver / virology*
  • Mice
  • Mice, Knockout
  • Scavenger Receptors, Class B / genetics
  • Scavenger Receptors, Class B / metabolism
  • Viral Envelope Proteins / genetics
  • Viral Envelope Proteins / metabolism
  • Virus Internalization*

Substances

  • Apolipoproteins E
  • Scarb1 protein, mouse
  • Scavenger Receptors, Class B
  • Viral Envelope Proteins
  • glycoprotein E2, Hepatitis C virus