Construction, screening and immunogenicity of the recombinant poxvirus vaccine rVTTδTK-RBD against SARS-CoV-2.
- Author:
Renshuang ZHAO
1
;
Yilong ZHU
2
;
Chao SHANG
3
;
Jicheng HAN
2
;
Zirui LIU
3
;
Zhiru XIU
2
;
Shanzhi LI
2
;
Yaru LI
1
;
Xia YANG
1
;
Xiao LI
4
,
5
;
Ningyi JIN
5
,
6
,
7
;
Xin JIN
8
;
Yiquan LI
6
,
9
Author Information
1. Preventive Veterinary Medicine Laboratory of Agricultural College, Yanbian University, Yanji 133000, China.
2. Academician Workstation of Jilin Province, Changchun University of Chinese Medicine, Changchun 130117, China.
3. Changchun Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Changchun 130122, China.
4. Academician Workstation of Jilin Province, Changchun University of Chinese Medicine, Changchun 130117
5. Changchun Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Changchun 130122, China.
6. Preventive Veterinary Medicine Laboratory of Agricultural College, Yanbian University, Yanji 133000
7. Academician Workstation of Jilin Province, Changchun University of Chinese Medicine, Changchun 130117
8. Preventive Veterinary Medicine Laboratory of Agricultural College, Yanbian University, Yanji 133000, China. *Corresponding authors, E-mail: jinxin@ybu.edu.cn.
9. Academician Workstation of Jilin Province, Changchun University of Chinese Medicine, Changchun 130117, China. *Corresponding authors, E-mail: liyq01@ccucm.edu.cn.
- Publication Type:Journal Article
- MeSH:
Animals;
Mice;
SARS-CoV-2/genetics*;
COVID-19;
Vaccines, Synthetic/genetics*;
Genes, Reporter;
Bacteriophages;
Mice, Inbred BALB C
- From:
Chinese Journal of Cellular and Molecular Immunology
2024;40(1):19-25
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a recombinant poxvirus vector vaccine, rVTTδTK-RBD, and to evaluate its safety and immunogenicity. Methods The receptor-binding domain (RBD) gene was synthesized with reference to the gene sequence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and was inserted into the polyclonal site of the self-constructed recombinant plasmid pSTKE, to construct the recombinant poxvirus shuttle vector pSTKE-RBD. This was then transfected into BHK-21 cells pre-infected with the vaccinia virus Tiantan strain (VTT). The recombinant poxvirus rVTTδTK-RBD was successfully obtained after several rounds of fluorescence phage screening. The effect of rVTTδTK-RBD on the body mass of BALB/c mice was detected after immunizing mice by intra-nasal vaccination. The levels of specific and neutralizing antibodies produced by rVTTδTK-RBD on BALB/c mice were analyzed after immunizing mice intramuscularly. The effect of rVTTδTK-RBD on T cell subsets in BALB/c mice was detected by flow cytometry. Results Through homologous recombination, enhanced green fluorescent protein (EGFP) screening marker, and multiple rounds of fluorescent phosphorescence phage screening, a recombinant poxvirus rVTTδTK-RBD, expressing RBD with deletions in the thymidine kinase (TK) gene, was successfully obtained, which was validated by PCR. The in vivo experiments on BALB/c mice showed that rVTTδTK-RBD was highly immunogenic against SARS-CoV-2 and significantly reduced toxicity to the body compared to the parental strain VTT. Conclusion The recombinant poxvirus vaccine rVTTδTK-RBD against SARS-CoV-2 is successfully constructed and obtained, with its safety and immunogenicity confirmed through various experiments.
