携带DEV-gB/NP28重组减毒沙门氏菌诱导鸭的免疫应答研究  

Study on the immune response of ducks induced by recombinant attenuated Salmonella carrying DEV-gB/NP28

作　　者：鲜思美[1,2] 李婷 冯将[1,2] 李鹏飞 包细明[1,2] 张益 文明[1,2] XIAN Si-mei;LI Ting;FENG Jiang;LI Peng-fei;BAO Xi-ming;ZHANG Yi;WEN Ming(College of Animal Science of Guizhou University, Guizhou 550025, China;Institude of Animal Disease Research of Guizhou, Guizhou 550025, China)

机构地区：[1]贵州大学动物科学学院,贵州贵阳550025 [2]贵州省动物疫病研究所,贵州贵阳550025

出　　处：《中国预防兽医学报》2018年第4期330-335,共6页Chinese Journal of Preventive Veterinary Medicine

基　　金：贵州省科技计划项目[黔科合(2016)2506号]; 国家自然科学基金项目(31260607); 贵州省动物疫病防控与兽医公共卫生保障科技创新人才团队[黔科合人才团队(2015)4016号]

摘　　要：为探究表达鸭肠炎病毒(DEV)gB/NP28双基因重组减毒沙门菌口服疫苗在鸭体内诱导免疫应答能力,本研究将构建的重组菌(pcDNA3.1-DEV-gB/NP28)/SL7207、DVE弱毒疫苗、重组质粒pcDNA3.1-DEV-gB/NP28及PBS分别免疫雏鸭,通过间接免疫荧光法检测免疫鸭脾脏、胸腺、法氏囊、哈德氏腺组织中的抗原表达;双抗体夹心ELISA方法检测肠道黏膜sIgA;间接ELISA方法监测血清中DEV特异性抗体、外周血T淋巴细胞增殖及IFN-γ、IL-4细胞因子含量,对该口服疫苗进行免疫学评价。结果表明,重组菌能够诱导鸭产生良好的黏膜免疫、体液免疫和细胞免疫应答;攻毒试验显示,在DVE强毒攻击下重组菌免疫可提供75%以上的保护,表明重组菌免疫对DEV攻击具有一定的免疫保护作用。To investigate the immune response of recombinant attenuated Salmonella typhimurium harbouring gB/NP28 double fusion genes of duck enteritis virus （DEV） as oral vaccine in ducks. Ducklings were respectively immunized with recombinant bacteria （pcDNA3.1-DEV-gB/NP28）/SL7207, DEV attenuated vaccine, recombinant plasmid pcDNA3.1-DEV-gB/ NP28 and PBS. Indirect immunofluorescence assay （IFA） was performed to detect the expression of antigen in ducks＇ spleen, thymus, bursa and Harder＇s gland. Double antibody, sandwich enzyme-linked immunosorbent assay （ELISA） was performed to detect the intestinal mucosa sIgA, and indirect ELISA was performed to detect the DEV-specific antibody in serum, peripheral blood T lymphocyte proliferation and IFN-γ, IL-4 cytokine contents in immunizated ducklings. These results indicated that the recombinant bacteria （pcDNA3.1-DEV-gB/NP28）/SL7207 could induce good mucosal, humoral and cellular immune response in ducks. The challenge experiment showed the recombinant bacteria could provide more than 75% protection after challenged with DEV and had good immune protection against DEV challenge.

关 键 词：鸭病毒性肠炎病毒 GB基因 NP28基因 减毒沙门氏菌 免疫应答 

分 类 号：S852.65[农业科学—基础兽医学]