Eukaryotic expression,purification and immunoefficacy evaluation of ferritin nan-oparticles of dengue virus type Ⅱ
10.16303/j.cnki.1005-4545.2024.06.05
- VernacularTitle:Ⅱ型登革病毒铁蛋白纳米颗粒蛋白的真核表达、纯化及免疫效果评价
- Author:
Junduo BAO
1
;
Xiangshu QIU
;
Yan GAO
;
Jiaqi ZHANG
;
Xiao LI
;
Xin JIN
;
Huijun LU
;
Ningyi JIN
Author Information
1. 延边大学农学院,吉林 延吉 133002
- Keywords:
dengue virus;
ferritin;
eukaryotic expression;
nanoparticles;
immunogenicity
- From:
Chinese Journal of Veterinary Science
2024;44(6):1120-1126
- CountryChina
- Language:Chinese
-
Abstract:
The E protein of dengue virus type Ⅱ was presented on ferritin of Helicobacter pylori to construct a novel dengue nanoparticle vaccine candidate,and the immunological indexes of the vac-cine were evaluated,aiming to provide new ideas for the development of dengue vaccine.The re-combinant plasmid of E-Ferritin was optimized and synthesized,and then transfected into HEK-293F cells.The recombinant protein was expressed,identified,purified and analyzed.Mice were im-munized with E-Ferritin nanoparticle vaccine by intramuscular injection on the hind limbs on the day 0,14 and 28.ELISA,neutralization test,flow cytometry and lymphocyte proliferation test were used to detect the levels of specific antibodies,neutralizing antibodies,CD3+,CD4+and CD8+T lymphocytes in spleen cells and the proliferation of spleen lymphocytes after specific stimulation.The target protein with a size of about 69 kDa was expressed in the cells with a single band.The purified protein concentration was 0.407 g/L,and the purity was 82.32％.The results from transmission electron microscopy showed that E-Ferritin protein could be recombined into a particle structure with a particle size of about 50 nm.The results of mouse immune experiments showed that E-Ferritin protein had good immunogenicity.The average specific antibody titer of E-ferritin protein in serum was 1∶92 160 after immunization 42 d.The main subclass of antibody was IgGl.The results of flow cytometry showed that E-Ferritin as an immunogen could induce higher levels of CD4+and CD8+T lymphocyte immune response.In lymphocyte proliferation test,the level of specific stimulation in the vaccine group was significantly higher than that in the non-specific stimulation group.In conclusion,the dengue virus envelope protein ferritin nanoparticle vaccine constructed in this study has good immunogenicity,which can provide reference for the de-velopment of new dengue vaccine candidates.
