Figure 6

The antiviral effect of IL-6 on HBV replication is not mediated through induction of IFNs. (A) Confluent 1.3ES2 cells were treated with or without 20 ng/ml of IL-6 for 2 day (day2) or 4 days (day4). Total RNA of each sample was extracted for the detection of IFN-α2b, IFN-β, IFN-γ and β-actin by RT-PCR analysis. Cells were transfected with 20 μg poly (I:C) for 6 h and served as the positive control for the induction of IFN-α2b and IFN-β simultaneously. Plasmid containing IFN-γ gene was served as the positive control for the detection of IFN-γ. (B) Confluent 1.3ES2 cells were treated with or without 20 ng/ml of IL-6 for 1 day (day1), 3 days (day3) or 6 days (day6). Total RNA of each sample was extracted for the detection of GBP-1 (target gene for IFN-γ), MxA (target gene for type I IFN) and GAPDH by RT-PCR analysis. Cells were treated with IFN-β or IFN-γ for 18 h and served as the positive control for the induction of GBP-1 and MxA genes. (C) Confluent 1.3ES2 cells were treated with various concentration of IL-6 or IFN-β for 4 days in the absence or presence of anti-IFN-β polyclonal antibody (400 U/ml). The level of HBV genome-containing nucleocapsids was measured as mentioned above. The expression level of β-actin was used as an internal control for sample loading.