A Hepatitis C virus genotype 1b posttransplant isolate with high replication efficiency in cell culture and its adaptation to infectious virus production in vitro and in vivo

Christian Heuss, Paul Rothhaar, Rani Burm, Ji Young Lee, Philipp Ralfs, Uta Haselmann, Luisa J. Ströh, Ombretta Colasanti, Cong Si Tran, Noemi Schä fer, Paul Schnitzler, Uta Merle, Ralf Bartenschlager, Arvind H. Patel, Frederik Graw, Thomas Krey, Vibor Laketa, Philip Meuleman, Volker Lohmann*

*Corresponding author for this work
3 Citations (Scopus)

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. Longterm 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 cellto- 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.

Original languageEnglish
Article numbere1010472
JournalPLoS Pathogens
Volume18
Issue number6
ISSN1553-7366
DOIs
Publication statusPublished - 06.2022

Research Areas and Centers

  • Academic Focus: Center for Infection and Inflammation Research (ZIEL)
  • Centers: Center for Structural and Cell Biology (CSCM/ZMSZ)

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