Real-time quantitative PCR analysis of human HepaRG cells infected with hepatitis E virus (HEV) [pre-array]

We aimed to identify interferon (IFN)-regulated genes that are differentially expressed during chronic HEV infection in human hepatocytes, the main site of HEV replication, using HepaRG cells. We have performed a preliminary screen using whole-cell RNA extracts prepared from HepaRG mock-infected or infected cells to determine whether HEV was able to trigger an IFN response. Different multiplicities of infection (MOI) (10 and 100 genome equivalent (GE)/cell) and time points (D+7, D+14, D+26, D+40, D+72 and D+100) were analyzed using the PCR array. We have also treated HepaRG cells after overnight treatment with IFN-β to confirm the ability of HepaRG to respond to IFN-I treatment. HepaRG cells infected with HEV-3f at a MOI of 10 or 100 GE/cell and their matched non-infected controls were analysed at different days post-infection. For MOI 10, 1 sample per time point was analysed. For MOI 100, 1 sample containing pooled RNA from triplicate samples was analysed.