Efficient Infectious Cell Culture Systems of the Hepatitis C Virus (HCV) Prototype Strains HCV-1 and H77

摘要: The first discovered and sequenced hepatitis C virus (HCV) genome the in vivo infectious HCV clones originated from prototype strains HCV-1 H77, respectively, both widely used research of this important human pathogen. In present study, we developed efficient cell culture systems for these genotype 1a by using HCV-1/SF9_A H77C clones. We initially adapted a with 5′UTR-NS5A (where UTR stands untranslated region) JFH1 NS5B-3′UTR (5-5A recombinant), including 2a-derived mutations F1464L/A1672S/D2979G (LSG), to grow efficiently Huh7.5 cells, thus identifying E2 mutation S399F. combination LSG/S399F reported TNcc(1a)-adaptive A1226G/Q1773H/N1927T/Y2981F/F2994S promoted adaptation full-length clone. An recombinant 17 (HCV1cc) replicated cells produced supernatant infectivity titers 104.0 focus-forming units (FFU)/ml. Eight were identified passaged viruses, A970T/I1312V/C2419R/A2919T essential particle production. Using CD81-deficient Huh7 further demonstrated importance A970T/I1312V/A2919T or A970T/C2419R/A2919T assembly that I1312V/C2419R played major role release. similar approach, found NS5B F2994R, here culture-adapted TN viruses common NS3 helicase (S1368P) derived viable 5-5A recombinants, initiated replication containing LSG mutations. harboring 19 (H77Ccc) spread after transfection subsequent infection naive reaching 103.5 104.4 FFU/ml, respectively. IMPORTANCE Hepatitis was 1989 cloning strain genome. 1997, two molecular other strain, shown be chimpanzees, but not vitro. hampered lack systems, which became available only 2005 discovery (genotype 2a), could establish cells. Recently, vitro (TN), 2a (J6), 2b (J8, DH8, DH10) key adaptive Globally, 1 is most prevalent. Studies H77 sequences have generated knowledge on HCV. Thus, will particular value advancing research. Moreover, our findings open new avenues isolates different genotypes.