我国D2-43株PrM-E基因的复制型载体质粒DNA的免疫原性研究  

Study on the immunogenity of replicative virus plas-mid DNA containing PrM-E gene of Chin ese D2-43virus strain

作　　者：陈水平[1] 秦鄂德[1] 于曼[1] 胡志君[1] 赵卫[1] 范宝昌[1] 王鹏程[1] 杨佩英[1] 

机构地区：[1]军事医学科学院微生物流行病学研究所,北京100071

出　　处：《细胞与分子免疫学杂志》2001年第5期466-467,481,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助;NO:39770036

摘　　要：目的通过观察含我国登革2型病毒株D2-43的PrM-E基因的复制型SFV重组质粒DNA的免疫原性,为登革新型疫苗的研制提供依据。方法将PrM-E基因自T载体上切下,插入复制型SFV病毒载体质粒DNA中。将此重组质粒DNA以电穿孔法导入BHK21细胞,表达产物的特异性用间接免疫荧光法进行鉴定。采用去除内毒素的质粒提取试剂盒制备重组质粒DNA,然后以不同剂量通过肌肉注射途径免疫Balb/c鼠,鼠血清中的抗体用间接免疫荧光法进行检测。结果含PrM-E基因的重组SFV质粒DNA在BHK21细胞中可表达登革2型病毒的特异蛋白;经免疫Balb/c鼠后,鼠血清可与登革D2-43感染的C6/36抗原片起特异的抗原抗体反应。结论含登革2型病毒PrM-E基因的复制型SFV病毒载体质粒DNA在Balb/c鼠中可诱导登革2型病毒特异抗体的产生,但抗体水平较低。Aim To study the immunogenity of replica tive plasmid DNA containing PrM -E gene of Chinese D2-43virus strain and lay a foundation for new vaccine of dengue.Methods The PrM -E gene was cut down from pGEM -T -ME with ApaI and ClaI and inserted into replicative SFV vector.The recombi nant plasmid DNA was electropo-rated into BHK21,then,expressed pr oducts were detected by IFA.Endotoxins -free plasmid DNA was pre pared with Endofree plasmid Maxi kit.Mice were immunized with different doses of the recombinant SFV plasmid DNA intramuscularly.The ti tres of antibody to dengue 2virus was measured by IFA.Results The recombinant SFV plasmid DNA coul d produce proteins specific to dengue virus in BHK21cells.And sera from mice,immunized with recombinant pl asmid,could react with antigen of dengue 2virus on microscope slides.Conclusion The recombinant SFV plasmid DNA,containing the PrM -E ge ne of D2-43virus strain,could produce specific antibody to d engue 2virus in mice,but the titre was low.

关 键 词：SFV载体 登革病毒 PrM-E基因 DNA免疫 

分 类 号：R373.33[医药卫生—病原生物学]