表达ApxIA的血清7型胸膜肺炎放线杆菌弱毒菌株的构建及特性分析  

Construction and characterization of Actinobacillus pleuropneumoniae serovar 7 live attenuated vaccine strain co-expressing ApxIA

作　　者：刘金林[1,2] 陈砚[1] 胡琳琳 贝为成[1] 陈焕春[1] 

机构地区：[1]华中农业大学农业微生物学国家重点实验室,武汉430070 [2]华中师范大学生命科学学院,武汉430079

出　　处：《生物工程学报》2010年第3期305-310,共6页Chinese Journal of Biotechnology

基　　金：国家自然科学基金项目(Nos.30970109,30600025,30530590);国家高技术研究发展计划(863计划)(No.2006AA10A206)资助~~

摘　　要：猪传染性胸膜肺炎是由胸膜肺炎放线杆菌引起的一种高度接触传染疾病,严重阻碍着全球养猪业的发展,疫苗接种是控制该病的有效措施。为提高胸膜肺炎放线杆菌弱毒疫苗的免疫效力,以及探索胸膜肺炎放线杆菌弱毒疫苗作为呼吸系统病原疫苗载体的可行性,通过穿梭质粒pJFF224-XN将完整的apxIA基因导入apxIIC基因缺失突变株HB04C-中,构建了含有apxIA和apxIIA基因的弱毒疫苗菌株HB04C2(apxIIC-/apxIIA+/apxIA+)。通过对HB04C2的生物学特性分析发现,穿梭质粒可稳定传代,并表达ApxIA,其生长特性未受穿梭质粒的影响。将HB04C2以气管接种方式免疫仔猪,可产生针对ApxIA和ApxIIA的抗体。二免后2周以高致病性的血清1型胸膜肺炎放线杆菌攻毒,该弱毒疫苗可提供良好的免疫保护效果。Actinobacillus pleuropneumoniae （A. pleuropneumoniae）, the causative agent of porcine contagious pleuropneumonia （PCP）, is a significant pathogen of the world pig industry, vaccination is potentially an effective tool for the prevention of PCP. The purpose of present study was to enhance the immunogenicity of A. pleuropneumoniae live vaccine strain HB04C-（serovar 7）, which was unable to express ApxIA, and to develop effective multivalent vaccines for the respiratory pathogens based on the attenuated A. pleuropneumoniae. We introduced a shuttle vector containing intact apxIA gene into HB04C-, generating HB04C2, an A. pleuropneumoniae serovar 7 live attenuated vaccine strain co-expressing ApxIA. Then we investigated the biological characteristics of HB04C2. We found that the shuttle vector expressing ApxIA was stable in HB04C2, and the growth ability of HB04C2 was not affected by the shuttle vector. We observed that HB04C2 elicited detectable antibodies against ApxIA and ApxIIA when it was administrated intratracheally as a live vaccine in pigs, and all immunized pigs were protected from heterologous virulent A. pleuropneumoniae （serovar 1） challenge. In conclusion, we demonstrated that A. pleuropneumoniae live vaccine could be used as a vector for expression of heterologous antigens.

关 键 词：胸膜肺炎放线杆菌 弱毒疫苗 多价疫苗 载体 

分 类 号：S852.61[农业科学—基础兽医学]