表达人高致病性禽流感病毒H5N1(安徽株)结构基因的DNA疫苗在小鼠中诱导的免疫应答分析  

DNA Vaccination via in vivo Electroporation Can Elicit Specific Immune Response against Highly Pathogenic H5N1 Influenza Viral Structural Antigens in Mice

作　　者：王文[1] 陈红[1] 谭文杰[1,2] 邓瑶[1] 王敏[1] 刘源[1] 殷霄[1] 张柯[1] 管洁[1] 周剑芳[1,2] 舒跃龙[1,2] 阮力[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所,北京100052 [2]中国疾病预防控制中心病毒基因工程重点实验室,北京100052

出　　处：《病毒学报》2010年第3期170-175,共6页Chinese Journal of Virology

基　　金："十一五"国家科技支撑计划项目(2006BAD06A15);国家重点研究发展计划(973)项目(2005CB523000);国家自然科学基金重大项目(30599433)

摘　　要：本研究旨在研发经济、高效的人高致病性禽流感病毒H5N1实验疫苗并优化免疫方案。首先优化并合成H5N1(安徽株)的血凝素基因(HAop)和神经氨酸酶基因(NAop)并证实其正确表达,再构建含有H5N1(安徽株)结构基因的多个单顺反子(HAwt、HAop和NAop)和双顺反子(HAop/M2和NAop/M1)DNA疫苗质粒。随后,采用不同途径的体内电转法在第0周和第3周2次免疫Balb/C小鼠,初步分析了抗原特异性体液免疫(HA血凝抑制抗体,NA特异性抗体,中和抗体)和细胞免疫应答(IFN-γELISpot)的特点。结果表明:含有优化血凝素基因(HAop)和优化神经氨酸酶基因(NAop)的DNA疫苗可以快速激发较强的免疫应答,尤其是细胞免疫应答;皮内电转所激发的体液免疫应答强于肌肉电转。本研究为新型H5N1疫苗的研发以及免疫方案的优化奠定了基础。This study aims to develop inexpensive and effective experimental vaccines against highly pathogenic H5N1 Avian Influenza （HPAI） virus and to optimize their immunization programs. To this end,we first synthesized the codon-optimized hemagglutinin gene （HAop） and neuraminidase gene （NAop）,both of which were derived from a H5N1 virus （Anhui strain）,and constructed successfully the DNA vaccines containing a single cistronic construct （HAwt,HAop,or NAop） or a bicistronic construct （HAop/M2 or NAop/M1） of H5N1 influenza virus origin. Their expression was confirmed by indirect immunofluorescent assay（IFA） and Western blotting. Then twice vaccination of mice with the DNA vaccines by injection intramuscularly or in vivo electroporation （EP） via two different routes was evaluated and analyzed by hemagglutination inhibition （HI） assay,NA-specific antibody detection,micro-neutralizing antibody test and IFN-γ ELISpot assay. Our results showed that the DNA vaccines with coden-optimized HAop and NAop constructs could quickly elicit a strong immune response by in vivo EP,especially the cellular immune response against HA and NA; the in vivo EP via intradermal route induced stronger humoral immune responses than those via intramuscular route. Our findings will pave a way for further development of novel DNA-based H5N1 vaccine and for the optimization of the immunization programs of DNA vaccine.

关 键 词：人高致病性禽流感病毒 H5N1 DNA疫苗 体内电转 

分 类 号：R373.1[医药卫生—病原生物学]