Research
Print page Print page
Switch language
The Capital Region of Denmark - a part of Copenhagen University Hospital
Published

Recombinant hepatitis C virus genotype 5a infectious cell culture systems expressing minimal JFH1 NS5B sequences permit polymerase inhibitor studies

Research output: Contribution to journalJournal articleResearchpeer-review

  1. Ribavirin inhibition of cell-culture infectious hepatitis C genotype 1-3 viruses is strain-dependent

    Research output: Contribution to journalJournal articleResearchpeer-review

  2. Production and characterization of high-titer serum-free cell culture grown hepatitis C virus particles of genotype 1-6

    Research output: Contribution to journalJournal articleResearchpeer-review

  3. Hepatitis C virus epitope exposure and neutralization by antibodies is affected by time and temperature

    Research output: Contribution to journalJournal articleResearchpeer-review

  4. Non-genotype-specific role of the hepatitis C virus 5' untranslated region in virus production and in inhibition by interferon

    Research output: Contribution to journalJournal articleResearchpeer-review

  1. Development of a downstream process for the production of an inactivated whole hepatitis C virus vaccine

    Research output: Contribution to journalJournal articleResearchpeer-review

  2. Global and local envelope protein dynamics of hepatitis C virus determine broad antibody sensitivity

    Research output: Contribution to journalJournal articleResearchpeer-review

  3. An alternate conformation of HCV E2 neutralizing face as an additional vaccine target

    Research output: Contribution to journalJournal articleResearchpeer-review

View graph of relations

The six major epidemiologically important hepatitis C virus (HCV) genotypes differ in global distribution and antiviral responses. Full-length infectious cell-culture adapted clones, the gold standard for HCV studies in vitro, are missing for genotypes 4 and 5. To address this challenge for genotype 5, we constructed a consensus full-length clone of strain SA13 (SA13fl), which was found non-viable in Huh7.5 cells. Step-wise adaptation of SA13fl-based recombinants, beginning with a virus encoding the NS5B-thumb domain and 3´UTR of JFH1 (SA13/JF372-X), resulted in a high-titer SA13 virus with only 41 JFH1-encoded NS5B-thumb residues (SA13/JF470-510cc); this required sixteen cell-culture adaptive substitutions within the SA13fl polyprotein and two 3´UTR-changes. SA13/JF372-X and SA13/JF470-510cc were equally sensitive to nucleoside polymerase inhibitors, including sofosbuvir, but showed differential sensitivity to inhibitors targeting the NS5B palm or thumb. SA13/JF470-510cc represents a model to elucidate the influence of HCV RNA elements on viral replication and map determinants of sensitivity to polymerase inhibitors.

Original languageEnglish
JournalVirology
Volume522
Pages (from-to)177-192
Number of pages16
ISSN0042-6822
DOIs
Publication statusPublished - 1 Sep 2018

    Research areas

  • Antiviral Agents/isolation & purification, Cell Culture Techniques/methods, Cell Line, Drug Evaluation, Preclinical/methods, Genotype, Hepacivirus/classification, Hepatocytes/virology, Humans, Viral Nonstructural Proteins/genetics, Virus Cultivation/methods

ID: 55377615