title: DNA vaccines against dengue virus type 2 based on truncate envelope protein or its domain III.
abstract: Two DNA vaccines were constructed encoding the ectodomain (domains I, II and III) of the DENV2 envelope protein (pE1D2) or only its domain III (pE2D2), fused to the human tissue plasminogen activator signal peptide (t-PA). The expression and secretion of recombinant proteins was confirmed in vitro in BHK cells transfected with the two plasmids, detected by immunofluorescence or immunoprecipitation of metabolically labeled gene products, using polyclonal and monoclonal antibodies against DENV2. Besides, results reveal that the ectodomain of the E protein can be efficiently expressed in vivo, in a mammalian system, without the prM protein that is hypothesized to act as a chaperonin during dengue infection. Balb/c mice were immunized with the DNA vaccines and challenged with a lethal dose of DENV2. All pE1D2-vaccinated mice survived challenge, while 45% of animals immunized with the pE2D2 died after infection. Furthermore, only 10% of pE1D2-immunized mice presented some clinical signs of infection after challenge, whereas most of animals inoculated with the pE2D2 showed effects of the disease with high morbidity degrees. Levels of neutralizing antibodies were significantly higher in pE1D2-vaccinated mice than in pE2D2-immunized animals, also suggesting that the pE1D2 vaccine was more protective than the pE2D2.
authors.affiliation: Laboratório de Biotecnologia e Fisiologia de Infecções Virais, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, Brazil.
authors.forename: Adriana S
authors.initials: AS
authors.lastname: Azevedo
authors.forename: Anna M Y
authors.initials: AM
authors.lastname: Yamamura
authors.forename: Marcos S
authors.initials: MS
authors.lastname: Freire
authors.forename: Gisela F
authors.initials: GF
authors.lastname: Trindade
authors.forename: Myrna
authors.initials: M
authors.lastname: Bonaldo
authors.forename: Ricardo
authors.initials: R
authors.lastname: Galler
authors.forename: Ada M B
authors.initials: AM
authors.lastname: Alves
mesh_terms: D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D003715:Dengue / Q000276:immunology; D003716:Dengue Virus / Q000276:immunology* / Q000472:pathogenicity; D005455:Fluorescent Antibody Technique; D006801:Humans; D047468:Immunoprecipitation; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids; D019444:Vaccines, DNA / Q000276:immunology*; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*
chemical_list: D019444:Vaccines, DNA; D014759:Viral Envelope Proteins