基因枪与肌肉注射猪繁殖与呼吸综合征病毒ORF5基因疫苗诱导小鼠体液及细胞免疫的比较研究  

Comparative Studies on the Humoral and Cellular Immune Responses of BALB/c Mice Immuned with pcDNA-PRRSV-ORF5 DNA Vaccine by Routes of Gene Gun Bombardment and Intramuscular Injection

作　　者：陈希文[1] 程安春[1] 汪铭书[1] 刘菲[2] 希尼尼根[1] 豆文波[1] 刘伍梅[1] 李雪梅[1] 张平英[1] 

机构地区：[1]四川农业大学动物科技学院,雅安625014 [2]动物疫病与人类健康四川省重点实验室,雅安625014

出　　处：《畜牧兽医学报》2005年第8期812-818,共7页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：四川省生物技术项目(01NG018-03);国家科技攻关重大项目(2004BA901A03);四川省重点建设学科项目(SZD0418)

摘　　要：猪繁殖与呼吸综合征病毒ORF5基因疫苗(pcDNA-PRRSV-ORF5)以不同免疫途径(基因枪和肌肉注射)免疫BALB/c小鼠,以PBS和空载质粒pcDNA3.1(+)为对照,采用流式细胞仪(FACS)、淋巴细胞增殖试验(MTT法)及间接ELISA试验分别对小鼠外周血中CD4+、CD8+T淋巴细胞数、T淋巴细胞的转化功能及小鼠血清中特异性PRRSV血清抗体IgG动态变化进行了检测。结果表明,pcDNA-PRRSV-ORF5基因疫苗接种小鼠后外周血对ConA有明显的反应性,试验组与对照组比较差异极显著(P<0.01)或显著(P<0.05),CD4+T淋巴细胞数在免疫后7d高于对照组(P<0.01),CD8+T淋巴细胞在免疫后28d高于对照组,不同途径基因疫苗接种小鼠后均诱导小鼠产生PRRSV特异性IgG。在诱导细胞免疫方面,基因枪和肌肉注射各组间无明显差异;在诱导体液免疫方面,基因枪法优于肌肉注射。研究表明制备的pcDNA-PRRSV-ORF5基因疫苗免疫小鼠能够诱导其机体产生良好的体液和细胞免疫应答,基因枪法较肌肉注射更能诱导体液免疫应答的产生。BALB/c mice were immunized with pcDNA-PRRSV-ORF5 DNA vaccine by different routes of gene gun bombardment and intramuscular injection, with PBS and pcDNA3. 1 （＋） as controls. Fluorescence activated cell sorter（FACS）was used to detect the number of CD4^＋ and CD8^＋ T lymphocytes. T lymphocyte proliferation test（MTT） and indirect ELISA methods were employed respectively to detect the proliferation of peripheral T lymphocyte and anti-PRRSV antibody. Peripheral T lymphocytes responses to ConA of the experiment group and the control group were significantly different （P〈0. 01）. In contrast with the control group, the number of CD4^＋ T lymphocyte 7 days after vaccination and CD8^＋ T lymphocyte 28 days after vaccination of the experiment group were significantly higher. The titers of specific IgG of the experiment group were higher than that of the control group. The results demonstrated that pcDNA-PRRSV-ORF5 DNA vaccine could induce good humoral and cellular immune response in mice. The titers of specific IgG of the group vaccinated by gene gun bombardment were higher than that by intramuscular injection.

关 键 词：基因枪 肌肉注射 猪 繁殖呼吸综合征 ORF5基因疫苗 小鼠 体液免疫 细胞免疫 

分 类 号：S858.28[农业科学—临床兽医学] S852.52[农业科学—兽医学]