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.
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...
| Main Authors: | , , , , , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2022-06-01
|
| Series: | PLoS Pathogens |
| Online Access: | https://doi.org/10.1371/journal.ppat.1010472 |
| _version_ | 1811155434169434112 |
|---|---|
| author | Christian Heuss Paul Rothhaar Rani Burm Ji-Young Lee Philipp Ralfs Uta Haselmann Luisa J Ströh Ombretta Colasanti Cong Si Tran Noemi Schäfer Paul Schnitzler Uta Merle Ralf Bartenschlager Arvind H Patel Frederik Graw Thomas Krey Vibor Laketa Philip Meuleman Volker Lohmann |
| author_facet | Christian Heuss Paul Rothhaar Rani Burm Ji-Young Lee Philipp Ralfs Uta Haselmann Luisa J Ströh Ombretta Colasanti Cong Si Tran Noemi Schäfer Paul Schnitzler Uta Merle Ralf Bartenschlager Arvind H Patel Frederik Graw Thomas Krey Vibor Laketa Philip Meuleman Volker Lohmann |
| author_sort | Christian Heuss |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-10T04:33:56Z |
| format | Article |
| id | doaj.art-7adc5d6f27254d5a8519af6c7a0be62e |
| institution | Directory Open Access Journal |
| issn | 1553-7366 1553-7374 |
| language | English |
| last_indexed | 2024-04-10T04:33:56Z |
| publishDate | 2022-06-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Pathogens |
| spelling | doaj.art-7adc5d6f27254d5a8519af6c7a0be62e2023-03-10T05:31:42ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742022-06-01186e101047210.1371/journal.ppat.1010472A 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.Christian HeussPaul RothhaarRani BurmJi-Young LeePhilipp RalfsUta HaselmannLuisa J StröhOmbretta ColasantiCong Si TranNoemi SchäferPaul SchnitzlerUta MerleRalf BartenschlagerArvind H PatelFrederik GrawThomas KreyVibor LaketaPhilip MeulemanVolker LohmannHepatitis 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.https://doi.org/10.1371/journal.ppat.1010472 |
| spellingShingle | Christian Heuss Paul Rothhaar Rani Burm Ji-Young Lee Philipp Ralfs Uta Haselmann Luisa J Ströh Ombretta Colasanti Cong Si Tran Noemi Schäfer Paul Schnitzler Uta Merle Ralf Bartenschlager Arvind H Patel Frederik Graw Thomas Krey Vibor Laketa Philip Meuleman Volker Lohmann 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 |
| title | 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. |
| title_full | 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. |
| title_fullStr | 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. |
| title_full_unstemmed | 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. |
| title_short | 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. |
| title_sort | 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 |
| url | https://doi.org/10.1371/journal.ppat.1010472 |
| work_keys_str_mv | AT christianheuss ahepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT paulrothhaar ahepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT raniburm ahepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT jiyounglee ahepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT philippralfs ahepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT utahaselmann ahepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT luisajstroh ahepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT ombrettacolasanti ahepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT congsitran ahepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT noemischafer ahepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT paulschnitzler ahepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT utamerle ahepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT ralfbartenschlager ahepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT arvindhpatel ahepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT frederikgraw ahepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT thomaskrey ahepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT viborlaketa ahepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT philipmeuleman ahepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT volkerlohmann ahepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT christianheuss hepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT paulrothhaar hepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT raniburm hepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT jiyounglee hepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT philippralfs hepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT utahaselmann hepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT luisajstroh hepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT ombrettacolasanti hepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT congsitran hepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT noemischafer hepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT paulschnitzler hepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT utamerle hepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT ralfbartenschlager hepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT arvindhpatel hepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT frederikgraw hepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT thomaskrey hepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT viborlaketa hepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT philipmeuleman hepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo AT volkerlohmann hepatitiscvirusgenotype1bposttransplantisolatewithhighreplicationefficiencyincellcultureanditsadaptationtoinfectiousvirusproductioninvitroandinvivo |