Highly efficient full-length hepatitis C virus genotype 1 (strain TN) infectious culture system

摘要: Chronic infection with hepatitis C virus (HCV) is an important cause of end stage liver disease worldwide. In the United States, most HCV-related associated genotype 1 infection, which remains difficult to treat. Drug and vaccine development was hampered by inability culture patient isolates representing HCV genotypes 1–7 subtypes; only a recombinant 2a genome (strain JFH1) spontaneously replicated in vitro. Recently, we identified three mutations F1464L/A1672S/D2979G (LSG) nonstructural (NS) proteins, essential for full-length (J6) 2b (J8) systems Huh7.5 cells. Here, developed highly efficient 1a TN) system. We initially found that LSG substitutions conferred viability intergenotypic composed TN 5′ untranslated region (5′UTR)-NS5A JFH1 NS5B-3′UTR; recovered viruses acquired two adaptive located NS3 NS4B. Introduction these changes into replication-deficient genome, harboring LSG, permitted production. Additional NS4B NS5B fully adapted virus. Thus, 8 (designated cell-culture derived, TNcc) efficiently released infectious particles ∼5 log10 focus-forming units per mL; passaged TNcc did not require additional changes. IFN-α directly acting antivirals targeting protease, NS5A, NS5B, each inhibited dose-dependently. Given unique importance pathogenesis, this system represents advance research. The approach used might permit other isolates, thus facilitating personalized treatment.