Efficient Initiation of HCV RNA Replication in Cell Culture

TLDR

Hepatitis C virus (HCV) infection is a global health problem affecting an estimated 170 million individuals worldwide. This work establishes a robust, cell‑based system for genetic and functional analyses of HCV replication. Multiple adaptive mutations in NS5A, including a single amino‑acid substitution that permits replication in 10 % of transfected hepatoma cells and a 47‑residue deletion in the interferon‑sensitivity determining region, markedly increase in‑vitro HCV replication, yet IFN‑α still rapidly inhibits replication independently of the ISDR.

Abstract

Hepatitis C virus (HCV) infection is a global health problem affecting an estimated 170 million individuals worldwide. We report the identification of multiple independent adaptive mutations that cluster in the HCV nonstructural protein NS5A and confer increased replicative ability in vitro. Among these adaptive mutations were a single amino acid substitution that allowed HCV RNA replication in 10% of transfected hepatoma cells and a deletion of 47 amino acids encompassing the interferon (IFN) sensitivity determining region (ISDR). Independent of the ISDR, IFN-α rapidly inhibited HCV RNA replication in vitro. This work establishes a robust, cell-based system for genetic and functional analyses of HCV replication.