title: [Immunogenicity and antigenicity of Japanese encephalitis virus envelope protein domain III].
abstract: To express the domain III gene of Japanese encephalitis virus (JEV) and to learn the possibility of developing the Dil protein as a subunit vaccine, we amplified the JEV DIII gene by PCR and constructed the expression plasmid pET-JE DIII by inserting JEV DIII gene into the prokaryotic expression vector pET-32a(+). The domain III protein of the attenuated strain SA14-14-2 was expressed as a thioredoxin (Trx) fusion protein, which was unique in forming a large fraction of the soluble recombinant protein. We immunized the rabbits and mice with the purified protein, tested the antigenicity and immunogenicity of JEV DIII protein by ELISA, Western blotting, plaque reduction test and observed the protective efficacy on challenged weanling mice with JEV. Rabbits immunized with the purified JEV DIII protein generated 1: 7 x 10(5) anti-JEV specific antibody titers. BALB/c mice immunized with the purified JEV E DIII protein generated 1: 8.2 x 10(4) anti-JEV specific antibody titers. And the neutralized antibody titer can reach 1:256, the survival rate of the immunized weanling mice was approximately 75%. Overall, this study highlighted that recombinant JEV E DIII protein delivered in mice and rabbits can generate high antibody titers against JEV, and protect some mice challenged with JEV. These studies can provide useful information for further developing the domain III recombinant protein as subunit vaccine against JEV.
authors.affiliation: Laboratory for Protein Engineering, Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Huang
authors.forename: Shan
authors.initials: S
authors.lastname: Liu
authors.forename: Peng
authors.initials: P
authors.lastname: Yang
authors.forename: Yun
authors.initials: Y
authors.lastname: Du
authors.forename: Zhiwei
authors.initials: Z
authors.lastname: Sun
authors.forename: Weiyuan
authors.initials: W
authors.lastname: Yu
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D004664:Encephalitis Virus, Japanese / Q000276:immunology*; D000939:Epitopes / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D007114:Immunization; D022321:Japanese Encephalitis Vaccines / Q000096:biosynthesis / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D017434:Protein Structure, Tertiary; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*
chemical_list: D000914:Antibodies, Viral; D000939:Epitopes; D022321:Japanese Encephalitis Vaccines; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins