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含CpG基元的SARS冠状病毒S-1基因片段DNA疫苗的构建及动物试验

Construction of DNA vaccine against S-1 gene fragments containing-CpG motif of SARS coronavirus in BALB/c mice and its application to animals

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【作者】 汤义何宏轩王冠胡慧中周凯段明星

【Author】 TANG Yi 1 , HE Hong-xuan 2 , WANG Guan 1 , HU Hui-zhong 1 , ZHOU Kai 3 , DUAN Ming-xing 1 (1. State Key Laboratory of Biomembrane and Membrane Biotechnology/Department of Biological Science and Biotechnology, Tsinghua University, Beijing 100084, China; 2.College of Veterinary Medicine, Cornell University, NY 14853,USA; 3.College of Life Science, Hebei Normal University, Shijiazhuang 050016, China)

【机构】 清华大学生物科学与技术系生物膜与膜生物工程国家重点实验室康乃尔大学兽医学院河北师范大学生命科学院清华大学生物科学与技术系生物膜与膜生物工程国家重点实验室 北京100084美国纽约14853北京100084河北石家庄050016北京100084

【摘要】 设计了3对含不同数目CpG基元(CpGmotifs)的引物,用SARS冠状病毒(SARSCoV)S1基因片段作为靶基因,通过PCR扩增出目的片段,插入真核表达载体pVAX1中。用电击法把构建好的不同质粒注入相应组BALB/c小鼠的股四头肌,2周后再加强免疫一次。每周分别测定血清IgG1和IgG2a抗体水平。结果,免疫组IgG1和IgG2a抗体浓度以及IgG2a与IgG1的比值均显著增加(P<0.001)。表明所构建的SARSCoVS基因S1片段的真核表达质粒可诱导小鼠产生很强的免疫反应。

【Abstract】 To construct the DNA vaccine against S-1 genefragments containing-CpG motif of SARS coronavirus and investigate immune responses in BALB/c mice, S-1 fragmentsof SARS-CoV S gene, containing different number of CpG motifs introduced by PCR, were inserted into eukaryotic expression vector pVAX1. The recombinant plasmid was injected into quadriceps femoris of BALB/c mice via electroporation and boosting with the same plasmid on week 2 after first immunization. IgG1 and IgG2a were measured weekly. The data indicated that the plasmid can elicit immune system to produce high levels of bothIgG1 and IgG2a significantly (P<0.001). The increased ratios of IgG2a to IgG1 indicated that they can elicitan intense Th1 immune response.The constructed eukaryoticexpression plasmid can induceintense immune response, which suggested that DNA vaccination may open up a new avenue for prevention against SARS.

【关键词】 SARS冠状病毒CpG基元DNA疫苗IgG2aIgG1
【Key words】 SARS coronavirusCpG motifsDNA vaccineIgG2aIgG1
【基金】 清华大学基础研究基金资助项目(03fd29)
  • 【文献出处】 中国兽医科技 ,Chinese Journal of Veterinary Science and Technology , 编辑部邮箱 ,2005年07期
  • 【分类号】R392
  • 【下载频次】106
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