Highly efficient JFH1-based cell-culture system for hepatitis C virus genotype 5a: failure of homologous neutralizing-antibody treatment to control infection

J Infect Dis. 2008 Dec 15;198(12):1756-65. doi: 10.1086/593021.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, CD
  • Cell Line
  • Genotype*
  • Hepacivirus / genetics*
  • Hepacivirus / physiology*
  • Humans
  • Immunoglobulin G / immunology
  • Mutation
  • Neutralization Tests
  • Scavenger Receptors, Class B / antagonists & inhibitors
  • Scavenger Receptors, Class B / metabolism
  • Tetraspanin 28
  • Time Factors
  • Viral Core Proteins / metabolism
  • Viral Nonstructural Proteins / metabolism
  • Virus Cultivation / methods*


  • Antigens, CD
  • CD81 protein, human
  • Immunoglobulin G
  • SCARB1 protein, human
  • Scavenger Receptors, Class B
  • Tetraspanin 28
  • Viral Core Proteins
  • Viral Nonstructural Proteins