我国登革2型病毒43株PrM-E基因的DNA免疫原性研究  

Study on the immunogenicity of the PrM-E gene DNA of Chinese dengue 2 virus 43 strain

作　　者：仝莉莉[1] 秦鄂德[1] 杨佩英[1] 于曼[1] 李同据[1] 欧武[1] 

机构地区：[1]军事医学科学院微生物学流行病学研究所

出　　处：《中华微生物学和免疫学杂志》2000年第4期301-305,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金!资助项目 (39770 0 36)

摘　　要：目的　观察我国登革 2型病毒 43株PrM E基因的DNA免疫原性及非甲基化细菌性CpG对DNA免疫效果的影响。方法　采用RT PCR和分子克隆技术构建PrM E基因片段 ,并将其插入真核表达载体pBK CMV中。在证明其可在哺乳动物细胞中高效表达的基础上 ,进一步用重组质粒DNA免疫小鼠。通过间接免疫荧光法对采集的鼠血清中的病毒特异抗体进行检测。结果　用含PrM E基因的重组质粒DNA免疫小鼠可诱导产生登革 2型病毒特异的抗体 ,且产生的抗体在小鼠体内可持续 3周以上。用含CpG的pUC19质粒和重组质粒的共免疫与单独免疫重组质粒所诱导的抗体水平没有显著差别。结论　我国登革 2型病毒PrM E基因重组质粒DNA具有一定的免疫原性。在本实验条件下 ,含非甲基化细菌性CpG的pUC19质粒DNA ,对PrM E基因的DNA免疫效果没有促进作用。Objective To study the immunogenicity of the PrM-E gene DNA of Chinese dengue 2 virus 43 strain and the effect of unmethylated CpG motif in the DNA immunization. Methods The PrM-E gene was constructed by RT-PCR and molecular cloning,and inserted into the eukaryotic expressive vector pBK-CMV.BALB/c mice were injected with the recombinant pCMV-ME plasmid DNA contained PrM-E gene interdermally and intramuscularly.The serum of immunized mice was detected for dengue specific antibody by indirect immunofluorescence. Results The high titer of antibody against D2-43 virus persisting for more than three weeks were detected in the immunized mice serum.It was not proved that the CpG of pUC19 could enhance the antigenicity of the recombinant plasmid pCMV-ME in this study. Conclusion The recombinant plasmid DNA of PrM-E gene but not unmethylated CpG was capable of inducting antibodies against D2-43 virus.

关 键 词：登革病毒 PrM-E基因 DNA免疫 非甲基化细菌性CpG 

分 类 号：R373[医药卫生—病原生物学]