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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 study the effects of novel adjuvant BW006 (CpG-containing oligonucleotide) and recombinant hepatitis B surface antigen (HBsAg) on the phenotype and function of mouse spleen dendritic cells (DCs). Methods Splenic DCs were stimulated with BW006 and / or HBsAg in vitro. The expression of CD80 and CD86 on DCs was detected by flow cytometry. The expression of IL-12p35 and IL-12p40 mRNA was detected by bDNA. The secretion of IL-12p70 was detected. Results The mRNA expressions of IL-12p35 and IL-12p40 reached the peak at 3 and 6 h after stimulation with 5 μg BW006, respectively, and increased by 1.2 and 26 folds respectively at 6 h and 6 h after stimulation. (74.53%, 36.10%). The level of IL-12p70 secreted by DCs reached the peak at 24 hours after stimulation (3 311.20 pg / ml). DCs were stimulated with 5μg BW006 and 40μg HBsAg for 24 hours. The positive rates of CD86 expression and CD86 expression were 25.17% and 51.65%, respectively. The secretion of IL-12p70 was 2699.70 pg / ml, which was not significantly different from that of BW006 stimulation group (P> 0.05). Conclusion BW006 has the function of early activation of DC, upregulation of CD80 and CD86 on DC surface and induction of IL-12 secretion. As a novel adjuvant for hepatitis B vaccine, BW006 has a good application prospect.