猪传染性胸膜肺炎新型亚单位菌苗对小鼠的效力研究  

Protective Efficacy of a Genetic Subunit Bacterin against Actinobacillus pleuropneumoniae in Mice

作　　者：刘建杰[1] 陈焕春[1] 何启盖[1] 吴斌[1] 李冲[1] 徐晓娟[1] 陈汉阳[1] 

机构地区：[1]华中农业大学动物医学院动物病毒室,武汉430070

出　　处：《畜牧兽医学报》2005年第2期177-180,共4页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(30200011);湖北省重点科技攻关项目(2001AA201B02)

摘　　要：利用大肠杆菌表达猪胸膜肺炎放线杆菌的分泌毒素ApxⅠ、ApxⅡ、ApxⅢ,提取表达产物包涵体,再加入我国流行的猪胸膜肺炎放线杆菌7型菌,和等量弗氏佐剂乳化后制成亚单位菌苗,免疫 BALB/c小鼠,间隔 2 周加强免疫1次。每次免疫后采血检测毒素Ⅰ、Ⅱ、Ⅲ的ELISA抗体和7型菌的血凝抗体,第2 次免疫 2 周后用 5LD50的猪胸膜肺炎放线杆菌1型、7型菌株进行攻毒。结果发现第2次免疫后抗体水平显著升高,用2种血清型进行攻毒后其保护力分别为83.3%和91.7%,从死亡小鼠的体内分离到了攻毒菌株,初步的动物试验表明此种新型亚单位菌苗对同型和异型的APP菌株具有较好的保护力,为进一步研制高效的亚单位菌苗打下基础。The exotoxin ApxⅠ, ApxⅡ and ApxⅢ of Actinobacillus pleuropneumoniae was expressed in E.coli, the recombinant protein was purified and mixed with A.pleuropneumoniae serotype 7, which is prevalent in China, then emulsified with Freund’s incomplete adjuvant in equal volumes to get a sort of mixed subunit bacterin. Groups of BALB/c mice were immunized twice at week 0 and 2,then challenged intraperitoneally with serotype 1 and serotype 7 at week 4. Antibodies were detected with ELISA and IHA. After second immunization, antibody level increased obviously and the immune protection rate (against) serotype 1 and serotype 7 reached 83.3% and 91.7% respectively, challenge strains were re-isolated from dead mice. These preliminary results showed that this mixed subunit bacterin had good protection (against) homologous and heterologous A.pleuropneumoniae,which built a good foundation for the further research of high efficacy vaccine against porcine pleuropneumonia.

关 键 词：亚单位 小鼠 免疫后 菌苗 保护力 加强免疫 大肠杆菌表达 猪胸膜肺炎放线杆菌 猪传染性胸膜肺炎 菌株 

分 类 号：S858[农业科学—临床兽医学] S852[农业科学—兽医学]