胸膜肺炎放线杆菌毒力因子重组蛋白免疫原性研究  

Study on immunogenic recombinant proteins of virulence factors of Actinobacillus pleuropneumoniae

作　　者：贺英[1] 赵萍[1] 储岳峰[1] 高鹏程[1] 石琴[1] 逯忠新[1] 

机构地区：[1]中国农业科学院兰州兽医研究所,家畜疫病病原生物学国家重点实验室,农业部畜禽病毒学重点开放实验室,甘肃兰州730046

出　　处：《西北农林科技大学学报（自然科学版）》2008年第10期14-18,共5页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家科技支撑计划子课题(2006BAD06A12)

摘　　要：【目的】研究猪传染性胸膜肺炎放线杆菌(APP)重组亚单位疫苗的免疫保护效果。【方法】用1.0mmol/LIPTG诱导重组表达质粒pET-ApxⅠA、pET-ApxⅢA和pPRoAppOML-1,SDS-PAGE鉴定。纯化的表达产物通过Western blot分析有较好的免疫原性。将rApxⅠ、rApxⅢ和rOmp组合作为疫苗Ⅰ组,rApxⅠ、rApxⅢ、rOmp和灭活菌苗组合作为疫苗Ⅱ组,APP 1、3、7型菌制备的灭活疫苗作为疫苗Ⅲ组,PBSⅣ作为空白对照组,进行家兔的免疫试验。免疫共分2次,每次间隔2周。【结果】疫苗Ⅱ组的抗体水平均显著高于疫苗Ⅲ组,疫苗Ⅰ组抗体水平不如疫苗Ⅲ组,其血清效价最高滴度为1∶256。【结论】猪传染性胸膜肺炎毒力因子重组蛋白具有一定的免疫原性,为进一步研究新型亚单位疫苗奠定了基础。【Objective】 The research studied immunoprotection effect of Actinobacillus pleuropneumonia recombination subunit vaccine.【Method】 The recombinants pET-ApxⅠA,pET-ApxⅢA,pPRoAppOML-1 were constructed by testing laboratory.Bacterial lyses prepared from 1.0 mmol/L IPTG induced cultures were loaded directly on to SDS-PAGE.The proteins were immunogenic when detected by Western-blot analysis.These results suggest that purified protein has bioactivity.The rabbit were immunized with recombinants of rApxⅠ,rApxⅢ and rOmp（group Ⅰ）;that of rApxⅠ,rApxⅢ,rOmp and viable organism（group Ⅱ）;inactivated vaccine of Actinobacillus pleuropneumonia（APP） serotype 1,3 and 7（group Ⅲ）;PBS（control group）.After detecting antibody titers,the immune protection efficacy of multicomponent recombinant subunit vaccines was assessed.【Result】 Results showed that the levels of antibodies,groupⅡwere significantly higher than those of the other two groups,but levels of antibodies of groupⅠwere lower than group Ⅲ.and sera titers reached the highest dilution of 1∶256.【Conclusion】 Recombinant proteins of virulence factors of Actinobacillus pleuropneumoniae have immunogenic,which is the basis to study new type subunit vaine.

关 键 词：猪传染性胸膜肺炎放线杆菌 免疫原性 基因工程苗 免疫试验 

分 类 号：S852.41[农业科学—基础兽医学] S859.797[农业科学—兽医学]