猪囊尾蚴病重组DNA疫苗pcDNA3.1-TSO45-4B在小鼠体内的表达及诱导的细胞免疫效应  

Expression of the recombinant DNA vaccine pcDNA 3.1-TSO45-4B against cysticercosis cellulosae and cell immunity response in mice

作　　者：王媛媛[1] 孙新[1] 方强[1] 胡守锋[1] 杨小迪[1] 张莺莺[1] 陈兴智[1] 王雪梅[1] 夏惠[1] 

机构地区：[1]蚌埠医学院病原生物学教研室安徽省感染与免疫重点实验室,安徽蚌埠233030

出　　处：《中国病原生物学杂志》2007年第4期262-265,共4页Journal of Pathogen Biology

基　　金：国家自然科学基金项目(No30600518);安徽省自然科学基金资助项目(No01043603);安徽省教育厅自然科学研究项目(No2001kj174)

摘　　要：目的观察基于六钩蚴期的猪囊尾蚴病基因疫苗pcDNA3.1-TSO45-4B在小鼠体内的表达及诱导的小鼠细胞免疫应答。方法将真核重组表达质粒pcDNA3.1-TSO45-4B肌注小鼠,RT-PCR方法扩增出目的条带;ELISA方法检测小鼠血清细胞因子IL-2、IL-4及IFN-γ含量。结果RT-PCR产物为单一条带,大小为456bp,与TSO45-4B大小一致。免疫小鼠第2周细胞因子含量达到较高水平,第4周达到最高水平,第8周开始下降。结论基于猪囊尾蚴保护性抗原TSO45-4B的DNA疫苗能在小鼠体内表达并有效诱导细胞免疫效应。Objective To investigate the expression in vivo of the recombinant DNA vaccine based on onchosphere stage-specific antigen TSO45-4B of the Taenia soliurn and the changes of IL-2, IL-4 AND IFN-？ of the immunized mice, Methods Mice were injected intramuscularly with the recombinant plasmid pcDNA 3, 1-TSO45-4B, and muscle samples of the mice were obtained in 48 h to 72 h, from which the mRNA of both TSO-45-4B and β-actin were amplified by RT-PCR with different primer respectively, the latter was as a system control, meanwhile cytokines IL-2, IL-4 and IFN- ？ in sera were detected by ELISA. Results The amplified product of the target gene was a single 456 bp band which showed the same molecular weight as the original, and all the cytokines detected were rised during the 2nd week, up to the highest level in the 6th week and then decreased in the 8th week. Conclusion The recombinant eukaryotie plasmid ocDNA 3.1-TSO45-4B expressed in vivo and the DNA vaccine could induce cytoimmunity response effectively in mice.

关 键 词：猪囊尾蚴 pcDNA 3 1-TSO45—4B 疫苗 RT—PCR 细胞免疫 

分 类 号：R383.24[医药卫生—医学寄生虫学]