猪胸膜肺炎放线杆菌毒素ApxⅠ对小鼠的致病性及免疫原性研究  

Study on Pathogenicity and Immunogenicity of Porcine Actinobacillus pleuropneumoniae ApxⅠtoxin

作　　者：马艳芳[1] 刁有祥[1] 张鑫[1] 纪巍[1] 孙法良[1] 孙宁[1] 

机构地区：[1]山东农业大学动物科技学院,山东泰安271018

出　　处：《中国农学通报》2009年第3期18-22,共5页Chinese Agricultural Science Bulletin

基　　金：山东省中青年科学家科研奖励基金"胸膜肺炎放线杆菌溶血毒素致病性小鼠动物模型的建立"(2007BS06004);山东农业大学科技创新基金"胸膜肺炎放线杆菌溶血毒素ApxⅠ;ApxⅡ基因突变株的构建与生物学特性"(23414)

摘　　要：将血清10型胸膜肺炎放线杆菌接种于含0.02%NAD、5%犊牛血清、1mMCaCl2的LB液体培养基中培养12h,离心后的上清液经硫酸铵盐析、葡聚糖凝胶分子筛层析,分离纯化ApxⅠ。将纯化的ApxⅠ进行半数致死量测定,并对小鼠剖检后进行内脏器官病理变化观察。将纯化的ApxⅠ与弗氏佐剂按1：1的比例混合制备亚单位疫苗,免疫小鼠,初免2周后加强免疫1次,二免2周后用血清10型胸膜肺炎放线杆菌攻毒,测定最佳免疫剂量。根据最佳免疫剂量于30日龄和45日龄免疫小鼠,60日龄时分别用血清1、3、5、7型胸膜肺炎放线杆菌攻毒。结果该毒素对小鼠的LD50为80.9mg/kg,LD50的95%平均可信限为63.2～103.5mg/kg;ApxⅠ亚单位疫苗的最佳免疫剂量为40μg,对血清1、3、5、7型胸膜肺炎放线杆菌的攻击具有一定的免疫保护效果。The Actinobacilluspleuropneumoniae serotype 10 strain isolated from Shandong province was cultivated in the LB medium containing 0.02%NAD, 5% Newborn Calf Sernm, lmM CaCl2 .The Apx I toxin was precipitated with ammonium sulfate and purified by chromatography on Sephadex G-200. In Pathogenic experiment, Karber＇ s method was used to test the LD50 of the Apx Ⅰ toxin in mice by intraperitioneal injection. In immune protective test, the purified Apx Ⅰ toxin was emulsified with Freund＇ s adjuvant and vaccinated the mice twice with a 2-week of interval. Two weeks after the second vaccination, the mice were challenged with the APP of serotype 10.The mice were immunized at 30 days and 45 days with the optimization immunizing dose, then the mice were challenged with the APP of serotype 1,3,5 and 7 at 60 days. The result revealed that the LD50 of the Apx Ⅰ toxin was 80.9 mg/kg, with a 95% confidence limit of 63.2-103.5 mg/kg; the Apx I toxin had a good immunogenicity and 40 μg per mouse was the optimization immunizing dose.

关 键 词：胸膜肺炎放线杆菌 毒素ApxⅠ 致病性 免疫原性 

分 类 号：S852.619[农业科学—基础兽医学]