新型CpG ODN对乙型肝炎病毒复制的抑制作用

In vitro study on the inhibitory effects of a novel CpG ODN on HBV replication

目的:探讨我室自行设计的CpG ODN(CpG BW001)在体外对乙型肝炎病毒(HBV)复制的抑制作用。方法:CpGBW001刺激人外周血单个核细胞(PBMC)48小时,用VSV病毒保护实验及ELISA法检测培养上清的病毒保护能力及其中的IFNα-含量。将上述CpG BW001刺激人PBMC的上清与HepG2.2.15细胞共孵育12天后收集培养上清,用放射免疫法检测该培养上清液中HBsAg及HBeAg的含量,用点杂交法检测HBV DNA的含量。结果:CpG BW001刺激人PBMC产生以IFNα-为主的抗病毒物质;CpG BW001刺激人PBMC的培养上清作用于HepG2.2.15细胞后,HepG2.2.15细胞培养上清中HBsAg和HBeAg的含量明显减少,且细胞内HBV DNA的含量也显著降低。结论:CpG BW001能通过刺激细胞产生抗病毒物质如IFNα-等,从而在体外抑制HBsAg和HBeAg的表达及HBV的复制。

Objective： To study the inhibitory effect of a novel CpG ODN （CpG BW001 ） on HBV replication in vitro. Methods： The anti-viral activity in the culture supernatant of CpG BW001-treated human PBMCs was detected by VSV protection bioassay and ELISA. HepG2.2.15 ceils were cultured with these supernatants and the supematants were harvested after incubation for 12 days. The levels of HBsAg and HBeAg in the supernatants were detected by radioimmunoassay and the level of HBV DNA in HepG2.2.15 cells was examined by dot blot. Results： CpG BW001 could stimulate human PBMC to product IFN-α and protect Vero cells from VSV attack. The levels of HBsAg and HBeAg produced by HepG2.2.15 cells and the level of HBV DNA in the supernatant of HepG2.2.15 cells were declined significantly when the HepG2.2.15 cells were cultured with the supernatant of human PBMC stimulated by CpG BW001. Conclusion： The novel CpG ODN （CpG BW001 ） is identified to inhibit the expression of HBsAg, HBeAg and to block the replication of HBV in vitro.