胸膜肺炎放线杆菌ApxⅡ蛋白重组腺病毒载体构建及免疫原性研究  

Construction and immunogenicity of recombinant adenovirus vector of Actinobacillus pleuropneumoniae ApxⅡprotein

作　　者：颜运秋[1] 黎满香[1] 赵枭健 龙娉[1] Yan Yunqiu;Li Manxiang;Zhao Xiaojian;Long Ping(College of Veterinary Medicine,Hu'nan Agricultural University,Hu'nan Changsha 410128)

机构地区：[1]湖南农业大学动物医学院,湖南长沙410128

出　　处：《现代畜牧兽医》2023年第11期21-25,共5页Modern Journal of Animal Husbandry and Veterinary Medicine

基　　金：湖南省教育厅优秀青年项目“湖南省猪传染性胸膜肺炎放线杆菌的分子流行病学调查及耐药性研究(20B29)”。

摘　　要：研究旨在构建胸膜肺炎放线杆菌ApxⅡ蛋白的复制缺陷型5型腺病毒载体的重组腺病毒。构建包含胸膜肺炎放线杆菌ApxⅡ基因的穿梭质粒pDC316-APXⅡ,并与腺病毒骨架质粒pBHGlox(delta)E13Cre共转染Hek293细胞,获得了重组腺病毒rAd-APXⅡ,将纯化后的重组腺病毒rAd-APXⅡ肌肉注射小鼠进行免疫原性分析。结果显示,试验成功构建了pDC316-APXⅡ质粒以及包装了rAd-APXⅡ病毒。rAd5-Null组未产生特异的APXⅡIgG抗体;低和高浓度rAd-APXⅡ组均可产生较高的APXⅡIgG抗体,其中高浓度组产生的抗体水平显著高于低浓度组(P<0.05);某疫苗组产生的抗体水平显著高于rAd-APXⅡ低浓度组(P<0.05),但低于rAd-APXⅡ高浓度组(P<0.05)。低、高浓度rAd-APXⅡ组和疫苗组的IgG1、IgG2a、IgG2b和IgG3抗体显著高于rAd-Null组(P<0.05);高浓度rAd-APXⅡ组和疫苗组抗体显著高于低浓度rAd-APXⅡ组(P<0.05)。研究表明,rAd-APXⅡ重组腺病毒免疫小鼠能够产生特异性抗体并显示良好的免疫原性。The aim of the study was to construct a recombinant adenovirus of replication-deficient type 5 adenovirus vector of Actinobacillus pleuropneumoniae ApxⅡprotein.The shuttle plasmid pDC316-APXⅡcontaining the ApxⅡgene of Actinobacillus pleuropneumonicus was constructed and co-transfected with the adenovirus skeleton plasmid pBHGlox(delta)E13Cre into Hek293 cells to obtain recombinant adenovirus rAd-APXⅡ.The immunogenicity of purified recombinant adenovirus rAd-APXⅡwas analyzed by intramuscular injection of mice.The results showed that pDC316-APXⅡplasmid was successfully constructed and rAd-APXⅡvirus was packaged.No specific APXⅡIgG antibody was produced in rAd5-Null group.Both low and high concentration rAd-APXⅡgroups produced higher APXⅡIgG antibodies,and the level of antibodies produced in high concentration group was significantly higher than that in low concentration group(P<0.05).The antibody level of one vaccine group was significantly higher than that of the low concentration group of rAd-APXⅡ(P<0.05),but lower than that of the high concentration group of rAd-APXⅡ(P<0.05).The IgG1,IgG2a,IgG2b and IgG3 antibodies in low and high concentration rAd-APXⅡand vaccine groups were significantly higher than those in rAd-Null group(P<0.05).The antibodies of high concentration rAd-APXⅡgroup and vaccine group were significantly higher than those of the low concentration rAd-APXⅡgroup(P<0.05).The study indicates that mice immunized with recombinant adenovirus rAd-APXⅡcan produce specific antibodies and show good immunogenicity.

关 键 词：猪传染性胸膜肺炎 胸膜肺炎放线杆菌 APXⅡ 重组腺病毒 免疫原性 

分 类 号：S855[农业科学—临床兽医学]