乙型肝炎疫苗新型佐剂BW006对小鼠脾树突细胞表型和功能的影响  

Influences of CpG-BW006,a Novel Adjuvant of Hepatitis B Vaccine,on Phenotype and Function of Murine Spleen Dendritic Cells

作　　者：张现臣[1] 何鹏[2] 胡忠玉[2] 邱少辉[2] 方鑫[2] 梁争论[2] 

机构地区：[1]北京民海生物科技有限公司,北京102600 [2]中国食品药品检定研究院,北京100050

出　　处：《中国生物制品学杂志》2011年第12期1396-1399,共4页Chinese Journal of Biologicals

基　　金：国家"十一五"传染病重大专项(2009ZX-10004-802)

摘　　要：目的研究新型佐剂BW006(含CpG基质的寡核苷酸)与重组乙型肝炎表面抗原(HBsAg)对小鼠脾树突细胞(Dendritic cell,DC)表型和功能的影响。方法应用BW006和/或HBsAg体外刺激小鼠脾DC,流式细胞仪检测DC膜表面分子CD80和CD86的表达水平,分支DNA(bDNA)法检测IL-12p35和IL-12p40 mRNA的表达水平,ELISA检测IL-12p70的分泌水平。结果 5μg BW006体外刺激DC 3和6 h,IL-12p35和IL-12p40的mRNA表达水平分别达峰值,刺激6 h时较刺激前分别增加了约1.2和26倍;刺激12和24 h,CD86和CD80的表达分别达峰值(74.53%和36.10%);刺激24 h,DC分泌IL-12p70的水平达高峰(3 311.20 pg/ml);5μg BW006联合40μg HBsAg体外刺激DC 24 h,DC表面分子CD80和CD86的表达阳性率分别为25.17%和51.65%,IL-12p70分泌水平达2 699.70 pg/ml,与BW006单独刺激组比较,差异均无统计学意义(P>0.05)。结论 BW006具有早期活化DC,上调DC表面协同刺激分子CD80和CD86表达及诱导IL-12分泌的功能,作为乙肝疫苗的新型佐剂,具有较好的应用前景。Objective To investigate the influence of BW006（the oligonucleotide containing GpG motif）,a novel adjuvant of hepatitis B vaccine,on the phenotype and function of murine spleen dendritic cells（DCs）.Methods Murine spleen DCs were stimulated with BW006 and / or HBsAg in vitro,and determined for expressions of CD80 and CD86 on surface by flow cytometry,for expression levels of IL-12p35 and IL-12p40 mRNAs by branch DNA（bDNA） method,and for secretion level of IL-12p70 by ELISA.Results After stimulation with 5 μg of BW006 in vitro for 3 ～ 6 h,expression levels of IL-12p35 and IL-12p40 mRNAs reached peak values,which increased by 1.2 and 26 folds respectively as compared with those before stimulation.However,the expression levels of CD86 and CD80 reached peak values 12 ～ 24 h after stimulation,which were 74.53% and 36.10% respectively.The secretion level of IL-12p70 reached a peak value of 3 311.20 pg / ml 24 h after stimulation.The expression positive rates of CD80 and CD86 on DC surface were 25.17% and 51.65% respectively 24 h after stimulation with 5 μg of BW006 and 40 μg of HBsAg in vitro,while the IL-12p70 secretion level reached 2 699.70 pg / ml,which showed no significant difference with those after stimulation with BW006 alone.Conclusion BW006 may activate DCs at early stage,up-regulate the expression of costimulatory molecules CD80 and CD86 on DC surface and induce IL-12 secretion,which is of a good prospect as a novel adjuvant of hepatitis B vaccine.

关 键 词：CPG-ODN 肝炎表面抗原 乙型 树突细胞 白细胞介素12 

分 类 号：R373.21[医药卫生—病原生物学] R392.33[医药卫生—基础医学]