In:
PLOS Pathogens, Public Library of Science (PLoS), Vol. 18, No. 6 ( 2022-6-28), p. e1010472-
Abstract:
Hepatitis C virus (HCV) is highly diverse and grouped into eight genotypes (gts). Infectious cell culture models are limited to a few subtypes and isolates, hampering the development of prophylactic vaccines. A consensus gt1b genome (termed GLT1) was generated from an HCV infected liver-transplanted patient. GLT1 replicated to an outstanding efficiency in Huh7 cells upon SEC14L2 expression, by use of replication enhancing mutations or with a previously developed inhibitor-based regimen. RNA replication levels almost reached JFH-1, but full-length genomes failed to produce detectable amounts of infectious virus. Long-term passaging led to the adaptation of a genome carrying 21 mutations and concomitant production of high levels of transmissible infectivity (GLT1cc). During the adaptation, GLT1 spread in the culture even in absence of detectable amounts of free virus, likely due to cell-to-cell transmission, which appeared to substantially contribute to spreading of other isolates as well. Mechanistically, genome replication and particle production efficiency were enhanced by adaptation, while cell entry competence of HCV pseudoparticles was not affected. Furthermore, GLT1cc retained the ability to replicate in human liver chimeric mice, which was critically dependent on a mutation in domain 3 of nonstructural protein NS5A. Over the course of infection, only one mutation in the surface glycoprotein E2 consistently reverted to wildtype, facilitating assembly in cell culture but potentially affecting CD81 interaction in vivo. Overall, GLT1cc is an efficient gt1b infectious cell culture model, paving the road to a rationale-based establishment of new infectious HCV isolates and represents an important novel tool for the development of prophylactic HCV vaccines.
Type of Medium:
Online Resource
ISSN:
1553-7374
DOI:
10.1371/journal.ppat.1010472
DOI:
10.1371/journal.ppat.1010472.g001
DOI:
10.1371/journal.ppat.1010472.g002
DOI:
10.1371/journal.ppat.1010472.g003
DOI:
10.1371/journal.ppat.1010472.g004
DOI:
10.1371/journal.ppat.1010472.g005
DOI:
10.1371/journal.ppat.1010472.g006
DOI:
10.1371/journal.ppat.1010472.g007
DOI:
10.1371/journal.ppat.1010472.g008
DOI:
10.1371/journal.ppat.1010472.s001
DOI:
10.1371/journal.ppat.1010472.s002
DOI:
10.1371/journal.ppat.1010472.s003
DOI:
10.1371/journal.ppat.1010472.s004
DOI:
10.1371/journal.ppat.1010472.s005
DOI:
10.1371/journal.ppat.1010472.s006
DOI:
10.1371/journal.ppat.1010472.s007
DOI:
10.1371/journal.ppat.1010472.s008
DOI:
10.1371/journal.ppat.1010472.s009
DOI:
10.1371/journal.ppat.1010472.s010
DOI:
10.1371/journal.ppat.1010472.s011
DOI:
10.1371/journal.ppat.1010472.s012
DOI:
10.1371/journal.ppat.1010472.s013
DOI:
10.1371/journal.ppat.1010472.s014
DOI:
10.1371/journal.ppat.1010472.s015
DOI:
10.1371/journal.ppat.1010472.s016
DOI:
10.1371/journal.ppat.1010472.s017
DOI:
10.1371/journal.ppat.1010472.s018
DOI:
10.1371/journal.ppat.1010472.s019
DOI:
10.1371/journal.ppat.1010472.r001
DOI:
10.1371/journal.ppat.1010472.r002
DOI:
10.1371/journal.ppat.1010472.r003
DOI:
10.1371/journal.ppat.1010472.r004
Language:
English
Publisher:
Public Library of Science (PLoS)
Publication Date:
2022
detail.hit.zdb_id:
2205412-1
