中国农业科学 ›› 2006, Vol. 39 ›› Issue (05): 1011-1017 .

• 畜牧·兽医·资源昆虫 • 上一篇    下一篇

猪繁殖与呼吸综合征病毒GP5和M蛋白共表达的自杀性DNA疫苗的构建及其免疫应答

江云波,方六荣,肖少波,张辉,陈焕春   

  1. 华中农业大学动物医学院
  • 收稿日期:2005-09-27 修回日期:1900-01-01 出版日期:2006-05-10 发布日期:2006-05-10
  • 通讯作者: 陈焕春

Construction and Immune Responses of the Suicidal DNA Vaccine Co-Expressing GP5 and M of Porcine Reproductive and Respiratory Syndrome Virus

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  1. 华中农业大学动物医学院
  • Received:2005-09-27 Revised:1900-01-01 Online:2006-05-10 Published:2006-05-10

摘要: 【目的】探讨猪繁殖与呼吸综合征病毒(PRRSV)自杀性DNA疫苗的免疫效果。【方法】将ORF5和ORF6基因分别插入可以同时表达两个外源基因的表达载体pSCA2的26S启动子下游,获得ORF5和ORF6双基因共表达自杀性DNA疫苗pSFV-56。【结果】Western blot检测证实ORF5和ORF6基因获得正确表达,并且所表达的GP5和M蛋白可以形成异源二聚体。将pSFV-56免疫Balb/c小鼠,首免后4周可检测到特异性PRRSV中和抗体,首免后8周的中和抗体最高可达1﹕32,同时还可检测到较高的特异性细胞免疫应答。进一步将pSFV-56免疫断奶仔猪,二免后4周即可产生1:8~1:16的中和抗体,直到二免后6周仍维持这种高水平的中和抗体,而且也可检测到较高的特异性细胞免疫应答。【结论】上述研究结果表明pSFV-56具有良好的免疫原性和诱发免疫动物产生较高免疫应答的能力。

关键词: “自杀性”DNA疫苗, 猪繁殖与呼吸综合征病毒, GP5和M蛋白共表达

Abstract: 【Objective】To investigate the immune efficiency of the suicidal DNA vaccines of porcine reproductive and respiratory syndrome virus (PRRSV). 【Method】The ORF5 and ORF6 genes were subcloned into the downstream of two independent subgenomic promoter 26S of a suicidal DNA vaccine expression vector pCSA2 resulting in the plasmid pSFV-56 co-expressing GP5 and M proteins.【Result】The result of Western blot showed that the GP5 and M proteins were expressed and formed disulfided-linked heterodimer. The suicidal DNA vaccine pSFV-56 was injected Balb/c mice and piglets to evaluate the induced immunological responses in vivo. The specific detectable anti-PRRSV neutralizing antibodies were produced in the vaccinated mice at 4 weeks post primary vaccination (PPV) and reached a peak 1:32 at 8 weeks PPV. The specific stronger cell-immunity responses were also observed. In addition, it was observed that the 1:8-1:16 neutralizing antibodies of the vaccinated piglets and the specific cell-immunity responses were produced.【Conclusion】The results of the present study indicated that the suicidal DNA vaccine pSFV-56 has well immunity and the capability inducing higher immune responses in the animals.

Key words: “Suicidal” DNA vaccine, porcine reproductive and respiratory syndrome virus (PRRSV), co-expressing GP5 and M