CpG-ODN体外抑制乙型肝炎病毒复制的研究

目的:探讨CpG-ODN体外对HBV复制和HBV抗原表达的抑制作用。方法:CpG-ODN体外刺激人外周血单个核细胞(PBMC),用ELISA检测培养液中IFN-α及IFN-γ的水平。将不同比例的PBMC培养上清与HepG2. 2. 15细胞共孵育 2、4、6、8d后,用ELISA和荧光定量PCR法,分别检测培养上清液中HB sAg、HBeAg和HBVDNA的水平。结果:CpG-ODN可诱导PB MC分泌IFN-α和IFN-γ。CpG-ODN本身虽不能直接抑制HBV的复制,但经刺激活的化的PBMC的培养上清,却能显著抑制 2. 2. 15细胞产生HBsAg、HBeAg和HBVDNA,在培养的第 8天,抑制率最高分别达 90. 8%、31. 3%和 32. 2%。结论:CpG-ODN作为一种新型的免疫调节剂,可诱发机体的免疫效应,抑制HBV复制和HBV抗原的表达。

Study on inhibitory effect of CpG-ODN on HBV replication in vitro

AIM: To study the inhibitory effect of synthetic oligodeoxynucleotides (ODN) containing unmethylated CpG dinucleotides (CpG-ODN) on hepatitis B virus(HBV) replication in vitro. METHODS: CpG-ODN was used to stimulate peripheral blood mononuclear cells(PBMCs). The IFN-alpha and IFN-gamma levels in the culture supernatant of stimulated PBMCs were measured by ELISA. The culture supernatant was added to HBV-transfected HepG2.2.15 cells, and after 2, 4, 6 and 8 days, HBsAg, HBeAg and HBV DNA levels in culture supernatant of 2.2.15 cells were detected by ELISA and quantitative PCR respectively. RESULTS: The CpG-ODN alone could not inhibit HBV replication, whereas the culture supernatant of PMBCs activated with CpG-ODN could remarkably inhibit the production of HBsAg, HBeAg and HBV DNA by 2.2.15 cells, reaching the maximal inhibitory rates of 90.8%, 31.3% and 32.2%, respectively on the 8th day. CONCLUSION: The study indicates that, as a novel immunoregulator, CpG-ODN can inhibit indirectly HBV replication in vitro.