tailieunhanh - Báo cáo khoa học: "Increased susceptibility of Huh7 cells to HCV replication does not require mutations in RIG-I"

Increased susceptibility of Huh7 cells to HCV replication does not require mutations in RIG-I | Feigelstock et al. Virology Journal 2010 7 44 http content 7 1 44 VIROLOGY JOURNAL RESEARCH Open Access Increased susceptibility of Huh7 cells to HCV replication does not require mutations in RIG-I Dino A Feigelstock 1 Kathleen B Mihalik1 Gerardo Kaplan2 Stephen M Feinstone1 Abstract Background The cytosolic retinoic acid-inducible gene I RIG-I is a pattern recognition receptor that senses HCV double-stranded RNA and triggers type I interferon pathways. The clone of human hepatoma Huh7 cells contains a mutation in RIG-I that is believed to be responsible for the improved replication of HCV in these cells relative to the parental strain. We hypothesized that in addition to RIG-I other determinant s outside the RIG-I coding sequence are involved in limiting HCV replication in cell culture. To test our hypothesis we analyzed Huh7 cell clones that support the efficient replication of HCV and analyzed the RIG-I gene. Results One clone termed Huh7D was more permissive for HCV replication and more efficient for HCV-neomycin and HCV-hygromycin based replicon colony formation than parental Huh7 cells. Nucleotide sequence analysis of the RIG-I mRNA coding region from Huh7D cells showed no mutations relative to Huh7 parental cells. Conclusions We derived a new Huh7 cell line Huh7D which is more permissive for HCV replication than parental Huh7 cells. The higher permissiveness of Huh7D cells is not due to mutations in the RIG-I protein indicating that cellular determinants other than the RIG-I amino-acid sequence are responsible for controlling HCV replication. In addition we have selected Huh7 cells resistant to hygromycin via newly generated HCV-replicons carrying the hygromycin resistant gene. Further studies on Huh7D cells will allow the identification of cellular factors that increased the susceptibility to HCV infection which could be targeted for anti-HCV therapies. Background Hepatitis C virus HCV infects nearly 200 million people worldwide 1 .