羊口疮病毒F1L与B2L基因核酸疫苗联合免疫小鼠特性的研究  

Study on immune response of DNA vaccine consisted with F1L and B2L gene in mice

作　　者：张友 杨倩 鲜思美[1,2] 吕碧英 饶体宇 吴伯梅 包涛涛 闵德省 ZHANG You;YANG Qian;XIAN Si-mei;Lü Bi-ying;RAO Ti-yu;WU Bo-mei;BAO Tao-tao;MIN De-sheng(College of Animal Science of Guizhou University,Guiyang 550025,China;Institude qf Animal Disease Research of Guizhou,Guiyang 550025,China;Animal.Disease Prevention and Control Center of Qixingguan,Bijie City,Guizhou,Bijie 551700,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 [3]贵州省毕节市七星关区动物疫病预防控制中心,贵州毕节551700

出　　处：《中国兽医科学》2020年第4期444-451,共8页Chinese Veterinary Science

基　　金：贵州省科技计划项目(黔科合支撑[2018]2264);贵州省科学技术基金项目(黔科合基础[2019]1113号);贵州省研究生教育创新计划项目(GZZ2017002)。

摘　　要：为评价羊口疮病毒(OrfV)F1L与B2L基因核酸疫苗PVAX1-F1L、PVAX1-B2L联合免疫小鼠诱导体液和细胞免疫的应答效果,分别将PVAX1-F1L+PVAX1-B2L、PVAX1-B2L、PVAX1-F1L、PVAX1空载体与PBS通过后腿肌肉注射方式免疫小鼠,采用ELISA方法检测免疫不同时间段的小鼠血清中OrfV特异性抗体及细胞因子IL-2、IL-4和IFN-γ,并通过MTT法检测免疫小鼠脾淋巴细胞增殖反应。结果显示,PVAX1-F1L+PVAX1-B2L联合免疫组小鼠血清抗体效价及IL-2、IFN-γ细胞因子水平显著(P<0.05)高于PVAX1-F1L和PVAX1-B2L单独免疫组,IL-4细胞因子水平差异不明显,但均与PVAX1空载体和PBS组差异极显著(P<0.01);PVAX1-F1L+PVAX1-B2L联合免疫组小鼠脾淋巴细胞增殖水平与PVAX1-F1L和PVAX1-B2L单独免疫组相比,差异显著(P<0.05),与PVAX1空载体和PBS组相比,差异极显著(P<0.01)。结果表明,PVAX1-F1L+PVAX1-B2L联合免疫小鼠能够增强OrfV特异性抗体的分泌和细胞免疫应答,这为羊口疮新型疫苗的研制奠定了理论基础。To evaluate the effect of inducing humoral and cellular immune response in mice immunized with DNA vaccines PVAX1-F1L and PVAX1-B2L of OrfV F1L and B2L gene,the test was divided into five groups PVAX1-F1L +PVAX1-B2L,PVAX1-B2L,PVAX1-F1L,PVAX1 empy vector and PBS group,they were respectively immunized by intramuscular injection in hind legs,The specific antibody of OrfV and cytokines IL-2,IL-4 and IFN-γ were detected by ELISA,and the proliferation of spleen lymphocytes was detected by MTT.The results showed that the ser-um antibody titer,IL-2 and IFN-γ cytokine levels of PVAX1-F1L+PVAX1-B2L combined immunization group were sign-ificantly higher than those of PVAX1-F1L and PVAX1-B2L group alone(P<0.05),There wa-s no significant difference in the level of IL-4,but it was significantly different from PVAX1 empt-y carrier and PBS group(P<0.01);the spleen lymphocyte proliferation level of PVAX1-F1L+PVAX1-B2L combined immunization group was significantly different from that of PVAX1-F1L a-nd PVAX1-B2L immunized groups(P<0.05),compared with PVAX1 empty vector and PBS gro-up.very significant(P<0.01).The results showed that PVAX1-F1L+PVAX1-B2L combined im-munization mice can enhance the secretion of OrfV-specific antibodies and cellular immune respo-nse,which provides a theoretical basis for the development of a new vaccine for Orf.

关 键 词：羊口疮病毒 F1L、B2L基因 核酸疫苗 免疫应答 

分 类 号：S852.659.1[农业科学—基础兽医学]