乙型肝炎病毒诱导肝细胞分泌的IL-35抑制病毒复制和病毒抗原分泌

Interleukin-35 induced by hepatitis B virus inhibits viral replication and viral antigen secretion in hepatocytes

目的研究乙型肝炎病毒(HBV)感染与肝细胞白细胞介素35(IL-35)表达的关系,并进一步探讨IL-35能否影响HBV复制及抗原分泌。方法转染含HBV全基因组(GenBank:GQ205440.1)1.3倍的复制子质粒或HBV编码基因表达质粒至HepG2细胞或LO2细胞以体外模拟HBV感染肝细胞,实时荧光定量PCR和Western blot法分析细胞内IL-35两个异源亚基EB病毒诱导基因3(EBI3)及P35 mRNA和蛋白含量。以(10、20、50)ng/mL重组人IL-35处理预转染pHBV1.3质粒的肝细胞,通过分析细胞上清液HBV-DNA水平及上清液乙型肝炎病毒调节蛋白X(HBx)、乙型肝炎病毒表面抗原(HBsAg)、乙型肝炎病毒核心抗原(HBcAg)和乙型肝炎病毒DNA聚合酶(HBp)的含量以评估IL-35对肝细胞HBV复制及抗原分泌的影响。结果转染pHBV1.3质粒的肝细胞内EBI3和P35的表达均较转染空载体pBLUE的肝细胞显著升高。除HBx、HBsAg、HBcAg和HBp均不能诱导肝细胞分泌IL-35。20~50 ng/mL重组人IL-35处理预转染pHBV1.3质粒的肝细胞后,细胞上清液HBV-DNA水平和病毒抗原含量显著降低。结论HBV感染促进肝细胞IL-35表达,而IL-35能抑制HBV复制和病毒抗原分泌。

Objective To explore the interaction between hepatitis B virus(HBV)infection and interleukin-35(IL-35)in vitro.Methods HepG2 or LO2 cells were transfected with HBV 1.3-fold replicator(GenBank:GQ205440.1)plasmid or HBV coding gene expression plasmid.The expression of IL-35 in hepatocytes was detected by both real-time PCR and Western blot analysis.HepG2 or LO2 cells transfected with pHBV1.3 plasmid were then treated with IL-35.The effects of IL-35 on HBV replication and antigen secretion were evaluated by analyzing the levels of HBV-DNA and virus secreting antigen in supernatant.Results The expression of IL-35 in hepatocytes with HBV infection was elevated markedly compared to hepatocytes without HBV infection.Moreover,upregulation of IL-35 expression in hepatocytes was induced by the HBx,but not HBs,HBc and HBp.In turn,IL-35 inhibited HBV replication in HepG2 and LO2 cells.Conclusion IL-35 induced by HBx has anti HBV activity in hepatocytes,which is a potential target for clinical treatment of chronic hepatitis B.