BACKGROUND: Recently, a hepatitis C virus (HCV) cell-culture system was developed that employed strain JFH1 (genotype 2a), and JFH1-based intra- and intergenotypic recombinants now permit functional studies of the structural genes (Core, E1, and E2), p7, and NS2 of genotypes 1-4. The goal was to adapt the system to employ genotype 5. METHODS: Huh7.5 cells infected with SA13/JFH1, containing Core-NS2 of strain SA13 (genotype 5a), were monitored for Core expression and for supernatant infectivity and HCV-RNA titers. Adaptive mutations of SA13/JFH1 were identified by sequence analysis of recovered genomes and reverse-genetic studies. Receptor blockage was performed with anti-CD81 and anti-SR-BI. For neutralization experiments, SA13/JFH1 or JFH1-based viruses of other genotypes were incubated with patient sera. RESULTS: SA13/JFH1 with NS2 and NS3 mutations yielded infectivity titers >10(5) TCID50/mL. Infection with SA13/JFH1 was inhibited by CD81 blocking and SR-BI blocking, respectively, and by preincubation with genotype 5a chronic-phase patient sera. Such sera had varying cross-genotype neutralization potential. However, preincubation and treatment with homologous neutralizing antibodies could not control SA13/JFH1 infection in culture. CONCLUSION: The SA13/JFH1 culture permits genotype 5a-specific studies of Core-NS2 function and interfering agents. The ability of HCV to spread in vivo during treatment with neutralizing antibodies was confirmed in vitro.
BACKGROUND: Recently, a hepatitis C virus (HCV) cell-culture system was developed that employed strain JFH1 (genotype 2a), and JFH1-based intra- and intergenotypic recombinants now permit functional studies of the structural genes (Core, E1, and E2), p7, and NS2 of genotypes 1-4. The goal was to adapt the system to employ genotype 5. METHODS: Huh7.5 cells infected with SA13/JFH1, containing Core-NS2 of strain SA13 (genotype 5a), were monitored for Core expression and for supernatant infectivity and HCV-RNA titers. Adaptive mutations of SA13/JFH1 were identified by sequence analysis of recovered genomes and reverse-genetic studies. Receptor blockage was performed with anti-CD81 and anti-SR-BI. For neutralization experiments, SA13/JFH1 or JFH1-based viruses of other genotypes were incubated with patient sera. RESULTS: SA13/JFH1 with NS2 and NS3 mutations yielded infectivity titers >10(5) TCID50/mL. Infection with SA13/JFH1 was inhibited by CD81 blocking and SR-BI blocking, respectively, and by preincubation with genotype 5a chronic-phase patient sera. Such sera had varying cross-genotype neutralization potential. However, preincubation and treatment with homologous neutralizing antibodies could not control SA13/JFH1 infection in culture. CONCLUSION: The SA13/JFH1 culture permits genotype 5a-specific studies of Core-NS2 function and interfering agents. The ability of HCV to spread in vivo during treatment with neutralizing antibodies was confirmed in vitro.
Authors: Yi-Ping Li; Santseharay Ramirez; Sanne B Jensen; Robert H Purcell; Judith M Gottwein; Jens Bukh Journal: Proc Natl Acad Sci U S A Date: 2012-11-14 Impact factor: 11.205
Authors: Jens Bukh; Philip Meuleman; Raymond Tellier; Ronald E Engle; Stephen M Feinstone; Gerald Eder; William C Satterfield; Sugantha Govindarajan; Krzysztof Krawczynski; Roger H Miller; Geert Leroux-Roels; Robert H Purcell Journal: J Infect Dis Date: 2010-05-01 Impact factor: 5.226
Authors: Zhenyong Keck; Wenyan Wang; Yong Wang; Patrick Lau; Thomas H R Carlsen; Jannick Prentoe; Jinming Xia; Arvind H Patel; Jens Bukh; Steven K H Foung Journal: J Virol Date: 2012-10-24 Impact factor: 5.103
Authors: Erica Silberstein; Kathleen Mihalik; Laura Ulitzky; Ewan P Plant; Montserrat Puig; Sara Gagneten; Mei-ying W Yu; Neerja Kaushik-Basu; Stephen M Feinstone; Deborah R Taylor Journal: PLoS Pathog Date: 2010-05-20 Impact factor: 6.823