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- Title
ENHANCED CYTOTOXICITY AGAINST THE TAB9 MINIGENE BY A PLASMID DNA-RECOMBINANT FOWLPOX VIRUS PRIME-BOOSTING REGIMEN AFTER PLASMID-BACKBONE ENGINEERING.
- Authors
Rodríguez, E. G.; Vázquez, D. M.; Herrera, A. M.; Quintana, D.; Abreu, N.; Duarte, C. A.
- Abstract
Background: Combined DNA priming-recombinant poxvirus boosting strategies have proven to be effective for generating antiviral immunity. In this work, we studied the effect of two different expression cassettes and the number of plasmid-backbone immunostimulatory sequences on the immune responses elicited by this vaccination regimen. Methods: The pAEC's transcriptional unit was replaced by another, containing the human cytomegalovirus promoter, a synthetic intron and synthetic RBG-based termination/polyadenilation sequences. Further constructs were generated by the insertion of 5, 10, and 20 copies of the 5'-AACGTT-3' immunostimulatory motif. Balb/c mice were immunized by intramuscular injection of 200 mg of each plasmid, coding for the HIV-1 multi-epitope TAB9. After three doses of DNA, a fourth boost with plasmid DNA or a TAB9-expressing recombinant Fowlpox virus (FPTAB9LZ) was administered. Immune responses were evaluated by ELISA and ELISPOT assays. Results: Serum IgG antibodies against TAB9 could not be detected. Only the animals receiving the new set of DNA constructs induced a detectable effector CTL response, which was further increased by the prime-boost regimen. The response was dependent on the number of immunostimulatory sequences. After in vitro re-stimulation with the IIIB peptide, significant CTL levels were evidenced for all the groups immunized with the new set of vectors. Conclusions: Plasmid backbone could influence the immune response in a combined DNA priming- viral boosting regime. The specific CTL response can be enhanced after mammalian transcriptional regulatory elements manipulation and the addition of an appropriated number of CpG motifs to plasmid backbone.
- Subjects
CELL-mediated cytotoxicity; CELL-mediated lympholysis; PLASMIDS; RECOMBINANT poxviruses; FOWL pox; SPINE
- Publication
Revista VacciMonitor (Vacunología y Temas Afines), 2002, Vol 11, Issue 4, p1
- ISSN
1025-028X
- Publication type
Article