欧洲型PRRSV重组腺病毒疫苗的构建及免疫效果评价

发布时间：2017-12-27 19:00

本文关键词：欧洲型PRRSV重组腺病毒疫苗的构建及免疫效果评价　出处：《中国病原生物学杂志》2015年11期 　论文类型：期刊论文

更多相关文章： EU-PRRSV 重组腺病毒 构建 免疫评价

【摘要】：目的构建欧洲型PRRSV重组腺病毒疫苗并对其免疫原性进行分析,为欧洲型PRRSV的感染预防提供依据。方法 PCR扩增欧洲型蓝耳病病毒ORF3、ORF5目的基因,将其连接到腺病毒穿梭载体,构建重组腺病毒穿梭质粒pacad-EU-ORF3、pacad-EU-ORF5、pacad-EU-ORF3-ORF5。将重组腺病毒穿梭质粒与骨架质粒共同转染至HEK 293细胞,构建重组腺病毒,并对构建的病毒用Western blot方法进行验证。用构建的重组腺病毒免疫BALB/c小鼠,通过淋巴细胞增殖试验对重组疫苗免疫效果进行初步评价。结果 Western blot显示重组腺病毒可稳定表达目的蛋白ORF3和ORF5,分子质量单位分别为30ku和22ku,与预期值相符。免疫小鼠淋巴细胞增殖试验显示重组腺病毒刺激指数均为PBS组1.5倍以上(P0.05),且rAd-EU-ORF3-ORF5组显著高于rAd-EU-ORF3组和rAd-EU-ORF5组,表明重组腺病毒疫苗能够诱导机体淋巴细胞增殖,增强机体的细胞免疫应答水平。结论成功包装出重组腺病毒rAdEU-ORF3、rAd-EU-ORF5和rAd-EU-ORF3-ORF5,动物实验显示其具有良好的免疫原性。
[Abstract]:Objective to construct a European type PRRSV recombinant adenovirus vaccine and analyze its immunogenicity to provide a basis for the prevention of infection in European type PRRSV. Methods the target genes of ORF3 and ORF5 were amplified by PCR, and then connected to adenovirus shuttle vector. The Recombinant Adenovirus Shuttle Plasmid pacad-EU-ORF3, pacad-EU-ORF5 and pacad-EU-ORF3-ORF5 were constructed. The Recombinant Adenovirus Shuttle Plasmid and skeleton plasmid were co transfected into HEK 293 cells, and then the recombinant adenovirus was constructed, and the constructed virus was verified by Western blot method. The recombinant adenovirus was used to immunise BALB/c mice, and the immune effect of the recombinant vaccine was evaluated by lymphocyte proliferation test. Results Western blot showed that recombinant adenovirus could stabilize the expression of target protein ORF3 and ORF5, and the molecular weight units were 30ku and 22ku, which were consistent with the expected value. Immunized mice lymphocyte proliferation test showed that the recombinant adenovirus stimulation index was more than 1.5 times for the PBS group (P0.05), and rAd-EU-ORF3-ORF5 group was significantly higher than that of rAd-EU-ORF3 group and rAd-EU-ORF5 group, showed that the recombinant adenovirus vaccine can induce lymphocyte proliferation in the body, enhance the cellular immune response of organism. Conclusion the recombinant adenovirus rAdEU-ORF3, rAd-EU-ORF5 and rAd-EU-ORF3-ORF5 were successfully packaged, and the animal experiments showed that the recombinant adenovirus had good immunogenicity.
【作者单位】：吉林农业大学动物科学技术学院;军事医学科学院军事兽医研究所;广西大学动物科学技术学院;
【基金】：国家863计划项目(No.2011AA10A208) 吉林省科技厅项目(No.20140309024NY)
【分类号】：R392
【正文快照】： ***ORF5,and rAd-EU-ORF3-ORF5were successfully constructed.Animal experiments indicated that the recombinant ade-novirus vaccines had good immunogenicity.猪繁殖与呼吸综合征(porcine reproductive andrespiratory syndrome,PRRS)是由猪繁殖与呼吸综合征病毒(porci

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