马疱疹病毒1型gB糖蛋白表位与小鼠C3d串联重组质粒免疫效果分析  

Analysis of Immune Effect of Recombinant Plasmid Which Contains Equine Herpesvirus Type 1 gB Glycoprotein Tandem Epitope and C3d of Mouse

作　　者：李静[1] 鲍子磊 范斌[1] 胡月[1] 宋焕堂 刘建华[1] 

机构地区：[1]新疆农业大学动物医学学院,乌鲁木齐830052

出　　处：《新疆农业大学学报》2017年第4期260-266,共7页Journal of Xinjiang Agricultural University

基　　金：新疆维吾尔自治区重大科技专项(2017A01002-2)

摘　　要：为了探究马疱疹病毒1型gB糖蛋白表位与小鼠C3d串联重组质粒的免疫效果。设计合成完整的抗原表位串联体B;以小鼠脾脏总RNA为模板,采用RT-PCR方法扩增出小鼠C3d基因;以EHV-1基因组为模板扩增出gB基因;构建重组质粒B-pVAX1、gB-pVAX1、C3d-pVAX1、B-C3d-pVAX1和gB-C3d-pVAX1;转染BHK-21细胞后能正常表达;将重组质粒免疫BALB/c雄性小鼠。结果表明:B-pVAX1免疫组抗体水平、IFN-γ和IL-4相对表达量均高于gB-pVAX1免疫组、C3d-pVAX1免疫组和对照(PBS)免疫组,差异极显著(P<0.01);B-C3dpVAX1免疫组抗体水平、IFN-γ和IL-4相对表达量均高于B-pVAX1免疫组、C3d-pVAX1免疫组和对照(PBS)免疫组,差异极显著(P<0.01);gB-C3d-pVAX1免疫组抗体水平、IFN-γ和IL-4相对表达量均高于gB-pVAX1免疫组、C3d-pVAX1免疫组和对照(PBS)免疫组,差异极显著(P<0.01);重组表位串联体B能够增强小鼠体液免疫和细胞免疫,同时C3d作为分子佐剂具有增强免疫的作用。It intends to explore the immune effect of the recombinant plasmid containing Equine herpesvirus type 1 gB glycoprotein tandem epitope and the C3 dof mouse.A complete antigen epitope tandem B was designed.C3 d was amplified by reverse transcriptase PCR(RT-PCR)using a total spleen RNA of mice as the template;gB was amplified by genome of EHV-1 as the template;The B-pVAX1,gB-pVAX1,C3 d-pVAX1,B-C3 d-pVAX1 and gB-C3 d-pVAX1 recombinant plasmid were constructed;The recombinant plasmid was transfected into BHK-21 cells with liposome transfection reagents 2000,and then BALB/c mice was immunized.The result showed that:B-pVAX1 immunized group antibody levels,IFN-γand IL-4 relative expression were higher than those of the gB-pVAX1 immunized group and the control group immunized by PBS C3 d-pVAX1,with a significant difference(P <0.01);B-C3 d-pVAX1 immunized group antibody levels and IFN-γand IL-4 relative expression were higher than the C3 d-pVAX1 immunized group and the control group immunized by PBS C3 d-pVAX1 with a significant difference(P <0.01);gB-C3 dpVAX1 immunized group antibody levels and IFN-γand IL-4 relative expression were higher than those of the gB-pVAX1 immunized group and the control group immunized by PBS C3 d-pVAX1,with a significant difference(P <0.01).Conclusion:Recombinant epitope B can enhance the humoral immunity and cellular immunity of mice and C3 das a molecular adjuvant has enhanced immune function.

关 键 词：马疱疹病毒1型 gB糖蛋白 B细胞表位 重组质粒 免疫效果 

分 类 号：S852.65[农业科学—基础兽医学]