中国农业科学 ›› 2007, Vol. 40 ›› Issue (4): 821-827 .

• 畜牧·兽医·资源昆虫 • 上一篇    下一篇

猪传染性胸膜肺炎重组亚单位疫苗在小鼠体内的免疫机制及其保护效力

邵美丽,刘思国,王春来,王 勇,宫 强,尹训南,郭设平,刘建东,佟恒敏   

  1. 中国农业科学院哈尔滨兽医研究所,兽医生物技术国家重点实验室
  • 收稿日期:2005-11-23 修回日期:1900-01-01 出版日期:2007-04-10 发布日期:2007-04-10
  • 通讯作者: 刘思国

Protective Efficacy and Immunologic Mechanism of the Recombinant Subunit Vaccine against Actinobacillus pleuropneumoniae in Mice

  1. 中国农业科学院哈尔滨兽医研究所,兽医生物技术国家重点实验室
  • Received:2005-11-23 Revised:1900-01-01 Online:2007-04-10 Published:2007-04-10

摘要: 【目的】欲构建具有较好交叉免疫保护效果的多组分重组亚单位疫苗,从而为猪传染性胸膜肺炎新型疫苗的研制提供参考。【方法】 利用分子克隆和重组表达技术获得了猪传染性胸膜肺炎放线杆菌6种主要毒力因子rApxI、rApxII、rApxIII、rApxIV、rApfa和rOMP。并设立以灭活苗(5×108cfu)为试验I组,以重组蛋白rApxI、rApxII、rApxIII和rOMP为试验II组,以rApxI、rApxII、rApxIII、rApxIV、rApfa和rOMP为试验III组,以PBS为空白对照组。分3次免疫BALB/c小鼠,每次间隔2周,第3次免疫后的第7天以APP1型(5×109cfu)和APP2型(5×1010cfu)进行攻毒保护试验。【结果】试验II组的4种抗体水平(ApxI、ApxII、ApxIII和OMP)和脾淋巴细胞诱生IL-2水平显著高于其它3组(P<0.05),脾淋巴细胞增殖水平也一直高于其它3组(P>0.05)。且试验II组对APP1型保护作用(9/10)明显高于试验I组(6/10)、试验III组(5/10)和对照组(0/10),而对APP2型保护作用(无肺脏损伤)也明显优于其它3组(典型肺部损伤),显示出细胞免疫和体液免疫水平与免疫攻毒保护之间存在着正相关性。【结论】试验II组通过激活体液免疫和细胞免疫,对不同血清型APP的攻击提供了很好的交叉保护作用。

关键词: 猪传染性胸膜肺炎, 重组亚单位疫苗, 保护力, 抗体, 淋巴细胞增殖, IL-2

Abstract: The study was focused on the comparison of protective efficacy between inactive vaccine (trial group I)and two recombinant subunit vaccines against serotype 1and serotype 2 of Actinobacillus pleuropneumoniae in mice, with one containing recombinant ApxI, ApxII, ApxIII and OMP(trial group II) and the other containing recombinant ApxI, ApxII, ApxIII, ApxIV, OMP and Apfa(trial group III). Then immunologic mechanism was determined preliminarily in mice on basis of humoral immunity and cell immunity. The BALB/c mouse were vaccinated at days 0 and 14 and 28, and then challenged with serotype 1(5×109cfu) and serotype 2 (5×1010cfu)intranasally at day 35. The results suggested that trial group II had a higher antibody level to four antigens and IL-2 production than others (P<0.05) .It had a slightly higher lymphocytes proliferation than others (P>0.05). The protective efficacy against serotype 1of trial group II (9/10) was obviously better than trial group I(6/10), trial group III(5/10)and control group(0/10), and the protective efficacy against serotype 2 of that (no lung lesion) was obviously better than others( typical lung lesion)., which demonstrated that antibody level had a positive correlation to protective efficacy. The study revealed that the recombinant subunit vaccine containing ApxI, ApxII, ApxIII and OMP provided significantly crossing protection by stimulating humoral immunity and cell immunity simutaneously.

Key words: porcine contagious pleuropneumonia, recombinant subunit vaccine, protective efficacy, antibody, lymphocytes proliferation, IL-2