Plasmid construction, expression, immunogenicity and protective efficacy of recombinant protein candidate vaccine of respiratory syncytial virus.
- Author:
Rui-Hong ZENG
1
;
Wei GONG
;
Xue-Ping FANG
;
Zhen-Ya ZHANG
;
Xing-Guo MEI
Author Information
1. Institute of Pharmacology and Toxicology, Academy of Military Medical Sciences, Beijing 100850, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Antibodies, Viral;
blood;
immunology;
Escherichia coli;
genetics;
metabolism;
Humans;
Immunoglobulin G;
blood;
immunology;
Mice;
Mice, Inbred BALB C;
Plasmids;
genetics;
immunology;
Recombinant Fusion Proteins;
biosynthesis;
genetics;
immunology;
Respiratory Syncytial Virus Infections;
prevention & control;
Respiratory Syncytial Virus Vaccines;
biosynthesis;
genetics;
immunology;
Respiratory Syncytial Virus, Human;
genetics;
immunology;
Viral Envelope Proteins;
genetics;
Viral Fusion Proteins;
genetics;
Viral Proteins;
genetics
- From:
Chinese Journal of Biotechnology
2005;21(4):534-539
- CountryChina
- Language:Chinese
-
Abstract:
To construct plasmid of recombinant protein candidate vaccine of respiratory syncytial virus, express it in E. coli, and to investigate its immunogenicity and protective efficacy. A CD8+ T cell epitope from respiratory syncytial virus (RSV) M2 protein F/M2:81 - 95 and the G:125-225 (G1) gene fragments from RSV-G protein containing B cell epitopes were amplified by PCR method and then inserted into the prokaryotic expression vector pET-DsbA after bonding to a linker. The fusion protein DsbA-G1-Linker-F/M2:81-95 (D-G1LF/M2) was expressed successfully in E. coli BL21 (DE3). The product was proved to be RSV-specific by Western-blot. After purified by affinity chromatography on Ni+ Sepharose and renatured by gradient dialysis. D-G1LF/M2 was used to immune BALB/c mice. D-G1LF/M2 induced high anti-D-G1LF/M2 IgG, anti-RSV IgG and neutralizing antibody titers in serum and lung of BALB/c mice, and elicied RSV-specific CTL responses. The IgG subclass distribution revealed that IgG1/IgG2a ratio was 2.66. Viral titration indicated that D-G1LF/M2 could protect BALB/c mice against RSV challenge in lung.
