CpG-ODN介导活化的人免疫细胞体外抗乙型肝炎病毒作用

Anti-hepatitis B virus effects of human peripheral blood mononuclear cells activated by CpG0ODN

目的:探讨CpG-ODN活化的人外周血单个核细胞(PBMC) 体外对HBV复制和表达的抑制作用. 方法:CpG-ODN体外刺激PBMC,ELISA测培养液IFN- α及IFN-γ的分泌;将CpG-ODN介导活化的PBMC与HepG2.2.15细胞按一定比例共孵育1 d、2 d和3 d后, ELISA检测培养上清液中HBsAg、HBeAg的分泌,荧光定量PCR检测HepG2.2.15细胞HBV DNA和HBV mRNA 的含量;并以MT7和酶学检测活化的PBMC对HepG2.2.15 细胞的杀伤作用. 结果:CpG-ODN有效诱导PBMC分泌IFN-α和IFN-γ; CpG-ODN本身虽不能直接抑制HBV的复制,但由CpG- ODN介导活化的PBMC却能显著减少HepG2.2.15细胞对HBsAg、HBeAg的分泌,同时对HBV DNA和HBV mRNA 的抑制作用亦明显增强;CpG-ODN介导活化的PBMC对HepG2.2.15的杀伤作用增强. 结论:CpG-ODN可通过活化机体免疫细胞,而具有明显的抗HBV复制和表达作用.

AIM: To investigate the inhibitory effect of peripheral blood mononuclear cells (PBMCs) activated by synthetic oligodeoxynucleotides (ODN) containing unmethylated CpG dinucleotides (CpG-ODN) on hepatitis B virus (HBV) in vitro. METHODS: CpG-ODN was co-cultured with PBMCs. The IFN-α and IFN-γ in the supernatant were measured by ELISA. PBMCs activated by CpG-ODN were added to HBV transfected HepG2.2.15 cells After 1, 2 and 3 days, HBsAg and HBeAg in the supernatant were measured by ELISA; HBV DNA and HBV mRNA in HepG2.2.15 cells were detected by fluorogenic quantitative PCR method. MTT method and enzyme assays were used to detect PBMC-mediated lytic activity against HepG2.2.15 cells. RESULTS: CpG-ODN induced high amounts of IFN-α as well as IFN-γ production (382.69±136.62, 37.42±6.55). Though CpG-ODN was unable to inhibit HBV replication directly, PMBC activated by CpG-ODN significantly reduced HBsAg and HBeAg secretion of HepG2.2.15 cells with rates of 82.6% and 52.4% at 72 h respectively (P<0.05). The remarkable inhibitory effects of PMBC activated by CpG-ODN on HBV DNA (21.5%) and HBV mRNA (81.3%) in HepG2.2.15 cells were also observed (P<0.05, P<0.01). CONCLUSION: PMBC activated by CpG-ODN can indirectly inhibit HBV replication and expression in vitro.