Preparation of the Coxsackievirus A16 VP1 subunit vaccine and detection of its immunogenicity
10.3760/cma.j.issn.0254-5101.2010.03.016
- VernacularTitle:手足口病CA16型VP1亚单位疫苗的构建制备及免疫原性初步分析
- Author:
Xiaonan LI
;
Deyan LUO
;
Zhongpeng ZHAO
;
Yueqiang DUAN
;
Peifeng LI
;
Xiliang WANG
- Publication Type:Journal Article
- Keywords:
Coxsackievirus A16 (CA16);
Bac-tu-Bac expression system;
Recombination VP1 protein;
Immunogenicity
- From:
Chinese Journal of Microbiology and Immunology
2010;30(3):250-255
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare VP1 protein vaccine of Coxsackievirus A16(CA16) and evalu-ate immunngenicity the subunit vaccines of Coxsackievirus (VP1), and to establish foundation for studying CA16 vaccine. Methods CA16 VP1 was amplified by RT-PCR and cloned into pFastBac HT A plasmid, recombinated with Bacmid DNA by transposition reaction and then transfected Sf9 cell, mixed with adjuvant AI(OH)_3. After immunization BALB/c mice, evaluating immune effectiveness after booster injections 2 weeks. Results The expressed protein was analyzed by SDS-PAGE and Western blot, mice immunized with CA16 (VP1) both induced specific IgG antibody and neutralization antibody. The best immunization antigen was 20 μg, IgG antibody was 1: 1600, neutralization antibody was 1:250, typical Th1/Th2 immune response was determined by lymphocyte proliferation assay and cytokine analysis. Conclusion The CA16 VP1 gene was cloned successfully and expressed in Sf9 insect cells, CA16 VP1 protein vaccine induced both humoral and cellular immune response, to lay solid foundation for further study on CA16 vaccine.
