Regulatory function of interferon‐inducible 44‐like for hepatitis B virus covalently closed circular DNA in primary human hepatocytes

Abstract Aim Curing hepatitis B virus (HBV) infection requires elimination of covalently closed circular DNA (cccDNA). Interferon (IFN)‐γ has noncytolytic antiviral potential; however, elimination of cccDNA could not be achieved. To enhance the regulatory effect, we comprehensively analyzed the host factors associated with cccDNA amplification and IFN‐γ and IFN‐α effects using an in vitro HBV infection system showing various transcription levels. Methods Primary human hepatocytes were infected with HBV using genomic plasmids carrying the basic core promoter mutation A1762T/G1764A and/or the precore mutation G1896A and treated with IFN‐γ and IFN‐α. Comprehensive and functional studies involving microarray and small interfering RNA analysis revealed the host factors related to cccDNA regulation. Results The HBV infection system reproduced the HBV life cycle and showed various propagation levels. Microarray analysis revealed 53 genes correlated with the cccDNA levels. Of the 53 genes, expression of IFN‐induced protein 44‐like ( IFI44L ) was significantly upregulated by IFN‐γ and IFN‐α. The anti‐HBV effect of IFI44L is exerted regardless of IFN‐γ or IFN‐α by inhibiting the activation of nuclear factor‐κB and signal transducer and activator of transcription 1 pathways. Conclusions Using the in vitro HBV infection system, an IFN‐inducible molecule, IFI44L, associated with cccDNA amplification, was identified. These results suggest an innovative molecular strategy for the regulation of HBV cccDNA by controlling a novel host factor, IFI44L.