GM-CSF和大肠杆菌鞭毛蛋白对口蹄疫灭活疫苗免疫效果的影响  

Effect of GM-CSF and Escherichia coli flagellin on immune efficacy of inactivated foot-and-mouth disease vaccine

作　　者：李亚宁[1] 雷白时[1] 李丽敏[1] 庞洪泽 赵款 张武超 郑世学[1] 袁万哲[1,2] LI Ya-ning;LEI Bai-shi;LI Li-min;PANG Hong-ze;ZHAO Kuan;ZHANG Wu-chao;ZHENG Shi-xue;YUAN Wan-zhe(College of Veterinary Medicine,Hebei Agricultural University,Baoding 071001,China;Hebei Veterinary Biotechnology Engineering Technology Innovation Center,Baoding 071001,China)

机构地区：[1]河北农业大学动物医学院,河北保定071001 [2]河北省兽医生物技术工程技术创新中心,河北保定071001

出　　处：《中国兽医科学》2022年第3期358-365,共8页Chinese Veterinary Science

基　　金：河北省现代农业产业技术体系羊产业创新团队专项(HBCT2018140204);河北农业大学精准畜牧学科群建设项目(1090064);河北省自然科学基金重点项目(C2019204219)。

摘　　要：为探究粒细胞-巨噬细胞集落刺激因子(GM-CSF)和大肠杆菌鞭毛蛋白(FliC)对口蹄疫(FMD)灭活疫苗免疫效果的影响,通过大肠杆菌表达系统成功表达了重组猪GM-CSF蛋白和重组FliC蛋白,之后进行了重组蛋白的纯化与复性。将制备好的重组蛋白与口蹄疫灭活疫苗混合后免疫小鼠,免疫后第4、7、10、14天采集小鼠血清,用间接ELISA检测VP1结构蛋白抗体水平,第14天无菌分离小鼠脾用qPCR检测脾细胞因子IL-2、IFN-γ、IL-4、IL-10的相对表达量。ELISA检测结果表明,免疫后第7、10、14天,GM-CSF佐剂组和FliC佐剂组的抗体水平明显高于FMD疫苗组,复合佐剂的抗体水平高于FMD疫苗组和其他佐剂组。q PCR结果显示,GM-CSF佐剂组、FliC佐剂组与复合佐剂组中IL-2、IL-4、IL-10的相对表达量显著高于疫苗组和PBS组。由此可见,重组FliC蛋白和重组猪GM-CSF蛋白可以提高FMD灭活疫苗的免疫应答水平,且重组蛋白联合使用的效果更佳。In order to investigate the effect of granulocyte-macrophage colony stimulating factor(GM-CSF)and Escherichia coli flagellin(Fli C)on the immune effect of inactivated foot-and-mouth disease(FMD)vaccine,recombinant porcine GM-CSF protein and recombinant Fli C protein were successfully expressed by E.coli expression system,and then the recombinant protein was purified and renatured.The prepared recombinant protein was mixed with inactivated FMD vaccine to immunize mice,and the serum of mice was collected on day 4,7,10,and 14 after immunization to detect VP1 structural protein antibody levels by indirect ELISA.The relative expression of splenic cytokines IL-2,IFN-γ,IL-4,and IL-10 was measured by q PCR in aseptically isolated mouse spleens on day 14.The results of ELISA showed that the antibody levels of GM-CSF adjuvant and Fli C adjuvant group were significantly higher than that of FMD vaccine group on the 7 th,10 th and 14 th day after immunization.The antibody level of compound adjuvant was higher than those of FMD vaccine group and other adjuvant groups.The results of q PCR showed that the relative expression levels of IL-2,IL-4 and IL-10 in GM-CSF adjuvant group and Fli C adjuvant combined adjuvant group were significantly higher than those in vaccine group and PBS group.In conclusion,recombinant Fli C protein and recombinant porcine GM-CSF protein can improve the immune response of inactivated FMD vaccine,and the combination of recombinant proteins is more effective.

关 键 词：GM-CSF FLIC 口蹄疫疫苗 

分 类 号：S852.659.6[农业科学—基础兽医学]