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Position: Home > Articles > 鸭特异性CpG ODNs的筛选与初步应用 China Poultry 2019 (19) 23-27

鸭特异性CpG ODNs的筛选与初步应用

作  者：
任肖;张姗;房立春;侯绍华
单  位：
中国农业科学院北京畜牧兽医研究所
关键词：
CpG ODN;鸭;佐剂;鸭坦布苏病毒病灭活疫苗
摘  要：
研究旨在以鸭外周血淋巴细胞为模型对设计的18条CpG ODNs进行筛选获得鸭特异性的CpG ODN,并与鸭坦布苏病毒病灭活疫苗(HB株)联用评价免疫效果.分别通过淋巴细胞增殖试验和实时荧光定量PCR测定CpG作用后的刺激指数和相关免疫基因的mRNA表达水平,从而确定最优的CpG ODN,后用阻断ELISA测定其与HB株灭活苗联用免疫北京鸭的抗体水平.结果显示,通过细胞增殖试验初筛发现刺激指数最高的CpG ODN为B-2、C-3和P-4;实时荧光定量PCR检测发现P-4刺激的外周血淋巴细胞中,IL-4、IL-6、IL-10和IFN-α以及TLR21 mRNA表达水平最高;P-4与灭活苗联用后,第14、28、42和56天的抗体阻断率PI值均显著高于对照组(P<0.05).以上结果表明,P-4对鸭外周血淋巴细胞具有较强的免疫刺激活性,与鸭坦布苏病毒病灭活苗联用可促进早期抗体的产生并提高抗体水平.
作  者：
REN Xiao;ZHANG Shan;FANG Lichun;HOU Shaohua;Institute of Animal Sciences CAAS;
关键词：
CpG ODN;;duck;;adjuvant;;duck Tembusu virus inactivated vaccine
摘  要：
The aim of this study was to screen 18 designed CpG ODNs using duck peripheral blood lymphocytes as a model to obtain a duck-specific CpG ODN, which was used with the duck Tembusu virus inactivated vaccine(HB strain) for preliminary evaluation. The optimal CpG ODN was determined by lymphocyte proliferation assay and real-time quantitative PCR to detect the stimulation index and mRNA expression level of related immune genes based on the treatment of CpG ODNs. And the antibody titer of the immunized Pekin ducks was then measured by blocking ELISA. The results showed that the CpG ODN with the highest stimulation index was found to be B-2, C-3 and P-4 through cell proliferation experiment. The mRNA expression levels of IL-4, IL-6, IL-10, IFN-α and TLR21 were the highest in peripheral blood lymphocytes under the stimulation of P-4. The ELISA results showed that the average value of percent inhibition(PI) of antibody were significantly higher than those of the control groups on day 14, day 28,day 42 and day 56 after the combination of P-4 and inactivated vaccine. The above indicated that P-4 had strong stimulating activity against duck peripheral blood lymphocytes, and the combination with the duck Tembusu virus inactivated vaccine could promote antibody production at early period and improve the antibody level.

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