增强猪传染性胸膜肺炎灭活苗对小鼠保护力的研究  

Evaluation of the augmented inactive vaccine against actinobacillus pleuropneumoniae in mice

作　　者：王勇[1] 刘思国[1] 王春来[1] 邵美丽[1] 刘建东[1] 宫强[1] 迟磊[1] 赵昆[1] 郭设平[1] 车广头 

机构地区：[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,黑龙江哈尔滨150001 [2]东北农业大学,黑龙江哈尔滨150030

出　　处：《中国预防兽医学报》2007年第6期442-447,共6页Chinese Journal of Preventive Veterinary Medicine

摘　　要：为了提高猪传染性胸膜肺炎灭活苗的保护力,本研究利用基因重组技术表达了猪传染性胸膜肺炎放线杆菌6种主要致病因子蛋白:rApxⅠ、rApxⅡ、rApxⅢ、rApxⅣ、rApfa和rOMP。设立试验Ⅰ组(灭活苗),试验Ⅱ组(灭活苗、rApxⅠ、rApxⅡ、rApxⅢ和rOMP),试验Ⅲ组(灭活苗、rApxⅠ、rApxⅡ、rApxⅢ和rApxⅣ),试验Ⅳ组(灭活苗、rApxⅠ、rApxⅡ、rApxⅢ和rApfa)和空白对照组(PBS),每组26只BALB/c小鼠,分3次免疫,每次间隔2周,每周检测血清抗体水平,在免疫后第2周、第4周和第5周,MTT法检测脾脏T细胞增殖水平及ELISA法测定IFN-γ分泌水平。在第3次免疫后的第7 d,将每组剩余20只小鼠平均分成2组,分别以APP1型菌(5×10~9 CFU)和APP7型菌(1×10^(11) CFU)进行攻毒保护试验。结果表明,向灭活苗中添加重组亚单位成份可以提高机体的细胞免疫和体液免疫水平,并能提高灭活苗的交叉保护率;试验Ⅱ组的ApxⅠ和OMP抗体水平以及细胞免疫水平均显著高于其它各组(p<0.05),对于APP1和APP7型菌的攻毒也提供了明显优于其它各组的保护结果,试验Ⅲ、Ⅳ组的细胞、体液免疫水平及保护率较试验Ⅰ组也有所提高。结果显示出ApxⅠ、OMP抗体水平和细胞免疫水平与攻毒保护率之间存在着正相关性,向灭活苗中加入重组蛋白成份可以提高疫苗的交叉保护,试验Ⅱ组通过激活体液免疫和细胞免疫,对两个血清型APP的攻击提供了很好的保护作用,从而为猪传染性胸膜肺炎新型疫苗的研制提供参考。In order to improve the protective efficacy of inactive vaccine against actinobacillus pleuropneumoniae, six recombinant proteins （rApx Ⅰ , rApxⅡ, rApx Ⅲ, rApx Ⅳ, rApfa and rOMP） were examined for their ability to enhance the effectiveness of the inactive vaccine of APP7. Groups of BALB/c mice were inoculated with inactive vaccine （group Ⅰ ）, recombinant subunit vaccine containing inactive vaccine, rApx Ⅰ, rApx Ⅱ, rApx Ⅲ and rOMP （group Ⅱ ）; inactive vaccine, rApx Ⅰ, rApx Ⅱ, rApx Ⅲ and rApx Ⅳ （group Ⅲ）; inactive vaccine, rApx Ⅰ, rApx Ⅱ, rApx Ⅲ and rApfa （group Ⅳ）, and PBS only （control group）, at day 0 and 14 and 28, respectively. Antibody level was tested weekly and T lymphocytes proliferation and IFN-7 production were examined at days 13, 27 and 34 post vaccination. The mice were challenged intranasally with serotype 1 （5 x 109 CFW） and serotype 7 （1 x 10H CFU） at day 35. The results showed that the group Ⅱ had a higher level of antibodies to Apx Ⅰ and OMP, T lymphocytes proliferation and IFN-7 production than others groups （P〈0.05）, and a better protection against challenges by serotype 1 and serotype 7. The levels of humoral immunity and cell immunity had a positive correlation to protective efficacy. These results demonstrated that supplement of recombinant proteins in inactive vaccine improved vaccine efficacy. The recombinant subunit vaccine containing inactive vaccine, rApx Ⅰ , rApx Ⅱ, rApx Ⅲ and rOMP provided significantly better protection by stimulating humoral immunity and cell immunity simultaneously.

关 键 词：猪传染性胸膜肺炎 重组亚单位菌苗 保护力 淋巴细胞增殖 

分 类 号：S852.4[农业科学—基础兽医学]