汉滩病毒核酸疫苗滴鼻免疫小鼠表达动力学研究  

Construction of hantaan virus DNA vaccine and its kinetics of gene expression in mice by intranasal immunization

作　　者：石永兵[1] 诸葛洪祥[1] 金东华[1] 许菊[1] 钱峰 张学光[3] 

机构地区：[1]苏州大学医学院病原生物学教研室,215007 [2]国家人类基因组南方研究中心 [3]苏州大学医学院免疫学教研室

出　　处：《江苏医药》2004年第6期425-427,M004,共4页Jiangsu Medical Journal

基　　金：国家自然科学基金 (批准号 :3 9970 65 4)

摘　　要：目的　构建汉滩病毒DNA疫苗 ,考查小鼠鼻内接种DNA疫苗后的基因组织定位及表达动力学 ,为进一步研究汉滩病毒DNA疫苗粘膜免疫的疗效作前期准备。方法　采用PCR法从PJSA1175 S质粒扩增汉滩病毒的S基因片段后装入PcDNA3 1B(- )载体 ,并以此候选DNA疫苗滴鼻免疫BALB/c小鼠 ,在不同时间、不同部位取其组织抽提基因组DNA和组织总RNA ,用PCR及RT PCR检测质粒的分布和表达动力学。结果　构建了汉滩病毒DNA疫苗PcDNA3 1B S1 3 。免疫接种该疫苗第 1天后在小鼠鼻咽、肺、肾、肝、脾、小肠等组织中均有质粒分布 ;第 1周内上述组织中均可检测到特异mRNA ,第 2周起小肠、肝及肾中特异mRNA消失。结论　成功构建了PcD NA3 1B S1 3 DNA疫苗 ,构建的疫苗滴鼻免疫后在小鼠体内得到广泛的分布及表达。Objective To construct DNA vaccine for hantaan virus S antigen and to study its kinetics of gene expression after intranasal immunization,prepared for further study of mucosal immune response by intranasal immunization with PcDNA3.1B-S 1.3.Methods S gene of hantaan virus was amplified by PCR from PJSA1175-S and cloned into DNA vaccine vector PcDNA3.1B(-).Mice were immunized by the constructed candidate for DNA vaccine.The location of DNA vaccine and the kinetics of gene expression were checked with PCR and RT-PCR.Results Hantaan virus DNA vaccine(PcDNA3.1B-S 1.3)was constructed.In the 1st day after intranasal immunization by the vaccine there were plasmids in nasal,lung,liver,spleen,gut and kidney.In the 1st week the special mRNA could be detected in all tissues above.From the 2nd week,the special mRNA couldnt be detected in gut,liver and kidney.Conclusion The DNA vaccine of hantaan virus was constructed successfully.The DNA vaccine and the special mRNA were detected widely in the intranasally immunized mice.

关 键 词：汉滩病毒 核酸疫苗 滴鼻免疫 小鼠 表达动力学 粘膜免疫 肾综合征出血热 HFRS 

分 类 号：R373[医药卫生—病原生物学] R-332[医药卫生—基础医学]