Expression of NA of influenza virus and C3d fusion gene in replication-defective recombinant adenovirus and its immune efficacy analysis
10.3760/cma.j.issn.1003-9279.2013.01.002
- VernacularTitle:流感病毒NA-C3d融合基因在非复制型重组腺病毒中的表达及其免疫效果的研究
- Author:
Feng HAN
1
;
Xuan WANG
;
Yan WANG
;
Xiao-Dong ZHAO
;
Shu-Hua WU
Author Information
1. 中国疾病预防控制中心病毒病预防控制所
- Keywords:
Influenza A virus;
Neuraminidase;
Adjuvants,immunologic;
Adenoviridae
- From:
Chinese Journal of Experimental and Clinical Virology
2013;27(1):5-7
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a replication-defective recombinant adenovirus expressing the fusion gene of neuraminidase (NA) gene in influenza virus A/FM/1/47 and C3d and to evaluate the induced immune efficacy.Methods NA-C3d was cloned into shutter vector pAdTrack-CMV,which was cotransformated with adenovirus DNA into E.coli BJ5183.The recombinant adenovirus genomic DNA was generated through homological recombination.The recombinant adenovirus was produced by transfecting 293 cell line with the genomic DNA and the induced immune efficacy in mice were analyzed.Results The integration of NA-C3d in the adenovirus genomic DNA and its expression were confirmed by PCR and Western-Blot assays respectively.After intranasal immunization,the serum IgG was induced at a titer of 1∶1000 and 1∶100 000 in BALB/c mice at primary and secondary immunization respectively.The vaccinated mice were completely survived when challenged with wide influenza virus.Conclusion recombinant adenovirus expressing NA-C3d was successfully constructed and it could induce desired immune efficacy.
