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• W33199758 abstract "Dengue is a disease caused by the dengue virus (DENV1-4). Despite severalstudies, no effective vaccine is yet commercially available. The envelope protein (E)of DENV is the viral surface major protein component, associated with numerousbiological activities. Thus, this protein is the main target for the induction of aprotective immune response based on neutralizing antibodies. In the present study,we evaluated the potential of DNA vaccines expressing the DENV2 E protein for theinduction of protection. Two plasmids were constructed, pE1D2 and pE2D2, whichcontain sequences encoding the ectodomain of the E protein (domains I, II and III) oronly its domain III, respectively, cloned upstream the encoding sequence of thehuman tissue plasminogen activator signal peptide (t-PA). Both plasmids mediatedexpression and secretion of the recombinant proteins in vitro in eukaryotic cells,detected with anti-DENV2 antibodies and evaluated by immunofluorescence ormetabolic labeling assay followed by imunopreciptation. Both DNA vaccines wereelicited neutralizing antibodies in Balb/c mice, with the highest antibody titersdetected in animals immunized with the pE1D2. The pE1D2 vaccine was also moreprotective in challenge tests with a lethal dose of DENV2, inducing 100% survival inimmunized mice, while 45% of animals vaccinated with the plasmid pE2D2 died afterinfection. Furthermore, 10% and 65% of the mice immunized with pE1D2 or pE2D2,respectively, showed morbidity after virus challenge. The vaccines pE1D2 andpE2D2 were also tested in combination with the chimeric YF17D-D2 virus, in a primeand booster system or with simultaneous immunizations. The YF17D-D2 chimericvirus was previously constructed by replacing the prM and E genes of 17DD yellowfever vaccinal virus with those from DENV2. The pE1D2 DNA vaccine combined withthe YF17D-D2 chimera induced high levels of neutralizing antibodies in animalsvaccinated with any of the different immunization schedules. Moreover, theseanimals showed 100% survival rates against a lethal challenge with DENV2, with noclinical signs of infection. The synergistic effect of combined immunization was alsoevident when we used the pE2D2 DNA vaccine and the YF17D-D2 virus, whichgenerated 100% survival in challenged animals. The cellular immune response wasevaluated by the production of IFN- by CD8+ T cells in ELISPOT assays, revealingthe activation of these cells in animals immunized with the pE1D2 alone or incombination with the YF17D-D2 chimera. Furthermore, analysis of the phenotypicprofile of CD4+ and CD8+ T cells showed low percentage of CD62L+ lymphocytes inanimals vaccinated with pE1D2, alone or combined with the chimeric virus, thusindicating that the DNA vaccine can influence the processes of T cell activation. NewDNA vaccines encoding the ectodomains of DENV1, 3 and 4 of the envelope protein(pE1D1, pE1D3 and pE1D4) were constructed and the expression of recombinantproteins was confirmed in vitro. These DNA vaccines will be further tested animalmodels." @default.
• W33199758 created "2016-06-24" @default.
• W33199758 creator A5090673357 @default.
• W33199758 date "2011-01-01" @default.
• W33199758 modified "2023-09-23" @default.
• W33199758 title "Desenvolvimento de vacinas de DNA contra o vírus da dengue baseadas na proteína do envelope viral" @default.
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