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猪GM-CSF对猪抗伪狂犬病病毒DNA疫苗的增强保护反应及免疫佐剂作用

Enhanced protective response and immuno-adjuvant effects of porcine GM-CSF on DNA vaccination of pigs against Aujeszky's disease virus.

作者信息

Somasundaram C, Takamatsu H, Andréoni C, Audonnet J C, Fischer L, Lefèvre F, Charley B

机构信息

Virologie et Immunologie moléculaires, INRA, Jouy en Josas, France.

出版信息

Vet Immunol Immunopathol. 1999 Sep 20;70(3-4):277-87. doi: 10.1016/s0165-2427(99)00085-9.

Abstract

This study was conducted to investigate whether the co-delivery of DNA encoding porcine cytokines would enhance a protective immune response in pigs to a Pseudorabies virus (PRV; or Aujeszky's disease virus) DNA vaccine. Aujeszky's disease in pigs results in respiratory and nervous symptoms with important economic losses. To evaluate cytokine effects, eukaryotic expression vectors were constructed for porcine GM-CSF, IL-2 and IFN-gamma. cDNA for each of these cytokines was inserted under the control of a CMV promoter in the pcDNA3 plasmid and cytokine expression was confirmed after DNA transfection in various mammalian cell cultures by bioassays (GM-CSF and IL2) and ELISA (IFN-gamma). Pigs were vaccinated by single intramuscular injection with plasmid DNA encoding PRV gB and gD along with various combinations of cytokine plasmid constructs. Pig serum was tested for the production of antibody by isotype specific anti-PRV ELISA. Pigs were then challenged with the highly virulent PRV strain NIA3 on day 21 after vaccination. The survival and growth rate of pigs were monitored for seven days after the viral challenge. The co-administration of GM-CSF plasmid increased the immune response induced by gB and gD PRV DNA vaccine. This immune response was characterized by an earlier appearance of anti-PRV IgG2, a significantly enhanced anti-PRV IgG1 and IgG2 antibody response, a significantly decreased and shortened viral excretion in nasal swabs and an improved protection to the viral challenge. In contrast, the co-administration of porcine IL-2 or IFN-gamma had no adjuvant effects. Our results thus demonstrate for the first time that the application of porcine GM-CSF gene in a DNA vaccine formulation can exert immuno-adjuvant and protective effects with single vaccination in the natural host pig against Aujeszky's disease.

摘要

本研究旨在调查编码猪细胞因子的DNA共同递送是否会增强猪对伪狂犬病病毒(PRV;或奥耶斯基氏病病毒)DNA疫苗的保护性免疫反应。猪的奥耶斯基氏病会导致呼吸和神经症状,并造成重大经济损失。为了评估细胞因子的作用,构建了猪GM-CSF、IL-2和IFN-γ的真核表达载体。将这些细胞因子各自的cDNA插入到pcDNA3质粒中CMV启动子的控制下,并通过生物测定法(GM-CSF和IL-2)和ELISA(IFN-γ)在各种哺乳动物细胞培养物中进行DNA转染后确认细胞因子表达。通过单次肌肉注射编码PRV gB和gD的质粒DNA以及细胞因子质粒构建体的各种组合对猪进行疫苗接种。通过同型特异性抗PRV ELISA检测猪血清中抗体的产生。然后在接种疫苗后第21天用高毒力PRV毒株NIA3对猪进行攻毒。在病毒攻毒后七天监测猪的存活和生长率。GM-CSF质粒的共同给药增强了gB和gD PRV DNA疫苗诱导的免疫反应。这种免疫反应的特征是抗PRV IgG2出现较早、抗PRV IgG1和IgG2抗体反应显著增强、鼻拭子中病毒排泄显著减少且持续时间缩短以及对病毒攻毒的保护作用改善。相比之下,猪IL-2或IFN-γ的共同给药没有佐剂作用。因此,我们的结果首次证明,在DNA疫苗制剂中应用猪GM-CSF基因可以在天然宿主猪中通过单次接种对奥耶斯基氏病发挥免疫佐剂和保护作用。

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