title: [Plasmid construction, expression, immunogenicity and protective efficacy of recombinant protein candidate vaccine of respiratory syncytial virus].
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.
authors.affiliation: Institute of Pharmacology and Toxicology, Academy of Military Medical Sciences, Beijing 100850, China.
authors.forename: Rui-Hong
authors.initials: RH
authors.lastname: Zeng
authors.forename: Wei
authors.initials: W
authors.lastname: Gong
authors.forename: Xue-Ping
authors.initials: XP
authors.lastname: Fang
authors.forename: Zhen-Ya
authors.initials: ZY
authors.lastname: Zhang
authors.forename: Xing-Guo
authors.initials: XG
authors.lastname: Mei
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids / Q000235:genetics / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D018357:Respiratory Syncytial Virus Infections / Q000517:prevention & control; D022261:Respiratory Syncytial Virus Vaccines / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D018113:Respiratory Syncytial Virus, Human / Q000235:genetics / Q000276:immunology*; D014759:Viral Envelope Proteins / Q000235:genetics*; D014760:Viral Fusion Proteins / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics*
chemical_list: D000914:Antibodies, Viral; C526850:F protein, human respiratory syncytial virus; D007074:Immunoglobulin G; C507355:M2-2 protein, respiratory syncytial virus; D011993:Recombinant Fusion Proteins; D022261:Respiratory Syncytial Virus Vaccines; D014759:Viral Envelope Proteins; D014760:Viral Fusion Proteins; D014764:Viral Proteins; C116204:attachment protein G