Evaluation of anti-HBV effect of recombinant human serum albumin-interferon α-2b fusion protein in vitro

OBJECTIVE To study the anti-HBV activity of prepared recombinant human serum albumininterferon α-2b fusion protein(HSA-IFNα-2b) in vitro.METHODS HepG2 cells were infected with recombinant adenovirus with green fluorescence protein and 1.6-fold HBV DNA(AdGFP-HBV).The expression of HBV antigens,HBsAg and HBeAg in culture medium was detected by ELISA assay.The toxicity of HSA-IFNα-2b on HepG2 cells was evaluated by MTT assay.The relative expression of HBV RNA in cells and the absolute quantity of HBV DNA in culture supernatant were determined by quantitative PCR assay.The activity of HBV enhancer Ⅰ was detected by Dual-Reporter gene assay.RESULTS HBV could replicate and express in HepG2 cells after infection with AdGFP-HBV.The expression of HBsAg and HBeAg in culture serum of HepG2 cells infected with AdGFP-HBV decreased by 51.32%(P 0.01) and 50.26%(P 0.01),respectively,when HSA-IFNα-2b 500 kU·L-1 was added.The same concentration of HSA-IFNα-2b didn't inhibit the proliferation of HepG2 cells,but inhibited HBsAg in a concentration-dependent manner.The regression formula between HBsAg inhibitory rate(Y) and concentration of HSA-IFNα-2b(X) was Y =21.11 lgX + 11.91(r2= 0.954),IC50= 63.76 kU ·L-1.HBV RNA in cells and HBV DNA in the culture serum decreased by 52.83%(P 0.01) and 53.07%(P 0.01),respectively,when HSA-IFNα-2b 500 kU·L-1 was added.The activity of enhancerⅠ decreased by 40.04%(P 0.01) when HSA-IFNα-2b 500 kU·L-1 was added.CONCLUSION The cell model of HBV replication for evaluating anti-HBV agents is successfully established.HSA-IFNα-2b exhibits noticeable anti-HBV effect in vitro.