中国农业科学 ›› 2006, Vol. 39 ›› Issue (8): 1651-1658 .

• 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基因对猪的DNA免疫

任慧英,郭鑫,杨汉春,王 芳,赵东升,杜希珍   

  1. 中国农业大学
  • 收稿日期:2005-08-08 修回日期:1900-01-01 出版日期:2006-08-10 发布日期:2006-08-10
  • 通讯作者: 郭鑫 guoxin guoxin

Immunization in Pigs Vaccined with DNA Encoding PRRSV GP5

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  1. 中国农业大学
  • Received:2005-08-08 Revised:1900-01-01 Online:2006-08-10 Published:2006-08-10

摘要: 【目的】检测PRRSV GP5 DNA重组质粒在猪体诱导的免疫反应以及细胞因子的佐剂效应。【方法】将表达PRRSV GP5的DNA重组质粒与表达3种细胞因子的重组质粒联合免疫SPF猪,经3次免疫后人工感染PRRSV,检测体液免疫、细胞免疫以及攻毒保护性反应。【结果】重组质粒pCI-GP5可诱导免疫猪产生中和抗体,效价达1:19.2,并可诱导机体产生较强的细胞免疫,攻毒后组织中PRRSV核酸的检出率下降39%,表明表达PRRSV GP5的DNA重组质粒pCI-GP5可诱导一定的免疫效力。pcDNA-IL-4与pCI-GP5共免可将中和抗体效价提高到1:35.5,且可增强细胞免疫反应,与对照组相比,病毒血症的出现频率减少17%,PRRSV阳性组织检出率下降64%;pcDNA-IL-2与pCI-GP5共免也可增强猪的细胞免疫,与对照组相比,病毒血症的出现频率下降50%,PRRSV阳性组织检出率减少59%; pcDNA-IFN- 与pCI-GP5共免对病毒血症的出现及病毒在组织中的分布均没有抑制作用。【结论】动物试验结果表明IL-4及IL-2可作为PRRSV GP5 DNA重组质粒的免疫佐剂。

关键词: 猪繁殖与呼吸综合征病毒, DNA重组质粒, 细胞因子, 免疫反应

Abstract: 【Objective】The aim is to analyze the immune responses and the effects of the cytokines induced by GP5 recombinant plasmids of PRRSV. 【Method】SPF piglets were immunized with DNA recombinant plasmids pCI-GP5 which expresses PRRSV GP5 by co-administration of pcDNA-IFN- , pcDNA-IL-2 and pcDNA-IL-4 for three inoculations. The vaccinated pigs were challenged intranasally with PRRSV BJ-4. 【Result】DNA immunization with pCI-GP5 resulted in the production of both neutralizing antibodies (1:19.2) and cellular immune responses two weeks after the third immunization. The pCI-GP5 showed significantly better protection from the PRRSV challenge compared with the control plasmid. The protection was characterized with a decrease of 39% PRRSV-positive tissues. In addition, the co-administration of pcDNA-IL-4 increased the immune response induced by pCI-GP5. This immune response was characterized by a significant enhancement of a lymphoproliferative response and a higher percentage of CD4-CD8+T cell, showing a higher viral neutralization titer of 1:35.5, a lower percentage of PRRSV-positive tissues and a significantly decreased frequency of viraemia (a drop of 64% and a drop of 17%, respectively, compared with the control group). Delivery of pcDNA-IL-2 paired with pCI-GP5 resulted in enhanced cellular immune responses, remarkably reduced percentage of PRRSV-positive tissues and frequency of viraemia (a drop of 59% and a drop of 50%, respectively, compared with the control group). In contrast, the co-administration of pcDNA-IFN- had no adjuvant effects. 【Conclusion】The results indicated that IL-4 and IL-2 could be used as adjuvant of PRRSV DNA vaccine.

Key words: PRRSV, DNA recombinant plasmid, cytokine, immune response