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目的比较用于乙型肝炎疫苗的3种寡聚脱氧核苷酸(CpG-oligodeoxynueleotide,CpG ODN)佐剂体外的免疫活性。方法分别将CpG ODN 684(沃森公司)、CpG ODN 1018(Dynavax公司)和CpG ODN 7909(Coley公司)序列体外刺激免疫人外周血单个核细胞(peripheral blood mononuclear cell,PBMC)和人树突状细胞系GEN,采用氚-胸腺嘧啶核苷(3H-TdR)渗入法检测人PBMC增殖活力,ELISA法检测GEN细胞中IL-6和IFNα水平。结果 CpG ODN 7909、CpG ODN 1018和CpG ODN 684诱导人PBMC的增殖水平明显高于未刺激的对照组,差异有统计学意义(P<0.000 1),其中CpG ODN 684和CpG ODN 7909序列诱导人PBMC的增殖水平相似,略高于CpG ODN 1018序列;3种CpG ODN在体外对免疫细胞均具有明显的活化作用,其中CpG ODN 684和CpG ODN 7909序列体外活化免疫细胞产生的IL-6和IFNα水平相似,但高于CpG ODN 1018序列。结论沃森公司CpG ODN 684序列体外活化免疫细胞功能与Coley公司CpG ODN 7909序列相似,略优于Dynavax公司CpG ODN 1018序列。
Objective To compare the immunological activities of three kinds of CpG oligodeoxynucleotides (CpG ODN) adjuvants used in hepatitis B vaccine in vitro. Methods Human peripheral blood mononuclear cells (PBMCs) and human dendritic cells (DCs) were stimulated in vitro with CpG ODN 684 (Watson), CpG ODN 1018 (Dynavax) and CpG ODN 7909 (Coley) Cell line GEN was used to detect the proliferative activity of human PBMCs by tritiated-thymidine (3H-TdR) infiltration method. The levels of IL-6 and IFNα in GEN cells were detected by ELISA. Results The proliferation of PBMC induced by CpG ODN 7909, CpG ODN 1018 and CpG ODN 684 was significantly higher than that of unstimulated control (P <0.000 1). CpG ODN 684 and CpG ODN 7909 induced human PBMC proliferated at a similar level and slightly higher than that of CpG ODN 1018. All three CpG ODNs had significant activation on immune cells in vitro. The CpG ODN 684 and CpG ODN 7909 sequences activated IL-6 and IFNα Similar in level, but higher than the CpG ODN 1018 sequence. Conclusions Watson’s CpG ODN 684 sequence has similar in vitro activation of immune cells to Coley’s CpG ODN 7909 sequence, slightly better than Dynavax’s CpG ODN 1018 sequence.