title: Japanese encephalitis subunit vaccine composed of virus-like envelope antigen particles purified from serum-free medium of a high-producer J12#26 cell clone.
abstract: A stable cell clone, J12#26, which continuously secretes large amounts of the envelope (E) antigen of Japanese encephalitis (JE) virus (J. Virol. 77 (2003) 8745) was adapted to serum-free medium. The J12#26 antigen possessed hemagglutinating activity, as well as the viral E and M proteins. More than 10 and 1mg of the antigen quantified with the licensed JE vaccine (JE-VAX) as a standard by E-ELISA and protein determination, respectively, were recovered from 500 ml of serum-free medium by membrane ultrafiltration, Sephacryl S-300 chromatography, sucrose gradient centrifugation and Sephadex G-25 chromatography. SDS-PAGE and Western blot analyses confirmed the high yield and purity of the J12#26 E antigen, which was comprised of small spherical virus-like particles (VLP) of approximately 25 nm in diameter. This antigen induced in mice without adjuvant neutralizing antibody (NT Ab) titers, as high as or higher than the licensed JE vaccine, and complete protection against challenge with wild-type virus. These results suggest that the J12#26 antigen is a promising second-generation JE subunit vaccine.
authors.affiliation: Department of Pathology, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan.
authors.forename: Eiji
authors.initials: E
authors.lastname: Mutoh
authors.forename: Toyokazu
authors.initials: T
authors.lastname: Ishikawa
authors.forename: Akihisa
authors.initials: A
authors.lastname: Takamizawa
authors.forename: Takeshi
authors.initials: T
authors.lastname: Kurata
authors.forename: Tetsutaro
authors.initials: T
authors.lastname: Sata
authors.forename: Asato
authors.initials: A
authors.lastname: Kojima
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000276:immunology; D002460:Cell Line; D002522:Chlorocebus aethiops; D002999:Clone Cells; D016895:Culture Media, Serum-Free; D004664:Encephalitis Virus, Japanese / Q000276:immunology* / Q000378:metabolism; D005260:Female; D022321:Japanese Encephalitis Vaccines / Q000096:biosynthesis / Q000191:economics / Q000276:immunology* / Q000302:isolation & purification; D051379:Mice; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*
chemical_list: D000956:Antigens, Viral; D016895:Culture Media, Serum-Free; D022321:Japanese Encephalitis Vaccines; D014759:Viral Envelope Proteins