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

Heuss, C. et al. (2022) A Hepatitis C virus genotype 1b post-transplant isolate with high replication efficiency in cell culture and its adaptation to infectious virus production in vitro and in vivo. PLoS Pathogens, 18(6), e1010472. (doi: 10.1371/journal.ppat.1010472) (PMID:35763545) (PMCID:PMC9273080)

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.

• Item Type: Articles
• Additional Information: Funding: This work was funded by grants from the Deutsche Forschungsgemeinschaft (DFG) LO1556/ 4-2 (278191845), TRR179 (272983813) and TRR209 (314905040) as well as a grant from the German Center for Infection Research (DZIF) (TTU 05.821) to VoL; grants from the Deutsche Forschungsgemeinschaft (DFG) TRR179 (272983813) and TRR209 (314905040) as well as grants from the German Center for Infection Research (DZIF) TTU 05.821 and TTU 05.712 to RBa; a UK Medical Research Council grant (MC_UU12014/2) to AHP, a Deutsche Forschungsgemeinschaft (DFG) grant (SFB 900, within project B10 (158989968)) to TK, a grant from the Chica and Heinz-Schaller Foundation to FG and grants from the Ghent University Special Research Fund (UGent BOF) as well as an Excellence of Science grant from the Research Foundation – Flanders (FWO) and FNRS to PM. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Patel, Professor Arvind

Creator Roles

Patel, A. H.: Resources, Writing – review and editing

• Authors: Heuss, C., Rothhaar, P., Burm, R., Lee, J.-Y., Ralfs, P., Haselmann, U., Ströh, L. J., Colasanti, O., Tran, C. S., Schäfer, N., Schnitzler, P., Merle, U., Bartenschlager, R., Patel, A. H., Graw, F., Krey, T., Laketa, V., Meuleman, P., and Lohmann, V.
• College/School: College of Medical Veterinary and Life Sciences > School of Infection & Immunity > Centre for Virus Research
• Journal Name: PLoS Pathogens
• Publisher: Public Library of Science
• ISSN: 1553-7366
• ISSN (Online): 1553-7374
• Published Online: 28 June 2022
• Copyright Holders: Copyright: © 2022 Heuss et al
• First Published: First published in PLoS Pathogens 18(6): e1010472
• Publisher Policy: Reproduced under a Creative Commons License

Project Code	Award No	Project Name	Principal Investigator	Funder's Name	Funder Ref	Lead Dept
172630		Basis of the host range and tissue tropism for hepatitis C virus	Arvind Patel	Medical Research Council (MRC)	MC_UU_12014/2	III - Centre for Virus Research