牛腐蹄病坏死梭杆菌白细胞毒素重组亚单位疫苗诱导小鼠免疫保护效果的观察  

Immune effect induced by the recombinant leukotoxin protein subunit vaccine of cattle foot rot Fusobacterium necrophorum strain H05 in mice

作　　者：郭东华[1] 王君伟[1] 孙玉国[1] 吕占军[1] 张建涛[1] 李林[1] 王洪斌[1] 

机构地区：[1]东北农业大学动物医学学院

出　　处：《中国预防兽医学报》2008年第5期398-402,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家科技攻关项目(2002BA518A04)

摘　　要：以大肠杆菌表达系统表达并纯化的牛源坏死杆菌H05菌株5个截短的重组白细胞毒素蛋白PL1(1aa～174aa)、PL2(311aa～644aa)、lktA3(1319aa～2026aa)、PL4(1960aa～2287aa)和PL5(3077aa～3241aa)为抗原分别免疫昆明系小白鼠,同时将灭活的天然坏死杆菌白细胞毒素和PBS作为免疫对照。ELISA结果显示,经4次免疫后5个重组蛋白和天然白细胞毒素均能介导小鼠产生抗白细胞毒素蛋白的特异性抗体,抗体效价分别为1∶6400、1∶12800、1∶6400、1∶12800、1∶3200和1∶800。免疫鼠活菌攻击观察结果和免疫鼠肝脏组织学变化结果一致证明,PL1、PL2、lktA3、PL4、PL5重组蛋白和天然白细胞毒素对小鼠均具有保护作用,5个重组蛋白的免疫保护作用均优于天然白细胞毒素,在5个重组蛋白中PL1、lktA3和PL4对鼠的免疫保护效果最好。Kunming mice were immunized with the purified recombinant leukotoxin proteins of PL1, PL2, 1ktA3, PL4 and PL5 emulsified in complete/incomplete Freund＇s adjuvant, respectively, and the native leukotoxin and PBS were also administrated as the positive and negative control. The sera were collected 3 days after the last booster inoculation for detecting specific antibodies against the recombinant proteins. The ELISA results showed that the antibody liters were 1：6 400, 1：12 800, 1：6 400, 1：12 800, 1：3 200 and 1：800 for the five recombinant proteins and native leukotoxin, respectively. The five recombinant proteins provided 100 % protection for mice against Fusobacterium necrophorum strain A25 challenging. However, the recombinant protein groups of PL1, 1ktA3, PL4 could provide better protection than that of other groups, comparing with the liver histological lesions.

关 键 词：腐蹄病 重组亚单位疫苗 免疫保护力 

分 类 号：S852.617[农业科学—基础兽医学]