Adapted J6/JFH1-Based Hepatitis C Virus Recombinants with Genotype-Specific NS4A Show Similar Efficacies against Lead Protease Inhibitors, Alpha Interferon, and a Putative NS4A Inhibitor

摘要: To facilitate studies of hepatitis C virus (HCV) NS4A, we aimed at developing J6/JFH1-based recombinants with genotype 1- to 7-specific NS4A proteins. We developed efficient culture systems expressing proteins genotypes (isolates) 1a (H77 and TN), 1b (J4), 2a (J6), 4a (ED43), 5a (SA13), 6a (HK6a), 7a (QC69), peak infectivity titers ∼3.5 4.5 log10 focus-forming units per ml. Except for growth depended on adaptive mutations identified in long-term culture. Genotype 1a, 1b, were adapted by amino acid substitutions F772S (p7) V1663A (NS4A), while 5a, 6a, required additional the NS3 protease and/or NS4A. demonstrated applicability study antivirals. 1 7 showed similar responses inhibitors telaprevir (VX-950), boceprevir (Sch503034), simeprevir (TMC435350), danoprevir (ITMN-191), vaniprevir (MK-7009), alpha interferon 2b, putative inhibitor ACH-806. The efficacy ACH-806 was lower than that not influenced changes acids 1042 1065 (in protease), which have been suggested mediate resistance replicons. viral spread under treatment ACH-806, without acquisition NS3-NS4A region. Relatively high concentrations inhibited assembly, but replication, a single-cycle production assay. HCV will benefitting from expression genotype-specific constant backbone context complete replication cycle, including functional evaluations