Enhanced cell-mediated IFN-γ-secreting activity against the HIV-1IIIB V3 peptide of the TAB9 multiepitope after DNA vaccine backbone engineering
| Autor: | Antonieta M. Herrera, Ernesto G Rodrı́guez, Dania M Vázquez, Carlos Duarte |
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| Rok vydání: | 2003 |
| Předmět: |
Transcription
Genetic Amino Acid Motifs Genetic Vectors Biophysics Cytomegalovirus Enzyme-Linked Immunosorbent Assay HIV Envelope Protein gp120 Biology Recombinant virus Biochemistry DNA vaccination law.invention Epitopes Interferon-gamma Mice Open Reading Frames chemistry.chemical_compound Plasmid law Escherichia coli Vaccines DNA Animals Cloning Molecular Promoter Regions Genetic Molecular Biology AIDS Vaccines Mice Inbred BALB C Models Genetic ELISPOT DNA Cell Biology Virology Molecular biology Introns Recombinant Proteins chemistry CpG site HIV-1 Recombinant DNA CpG Islands Peptides Spleen Plasmids Minigene |
| Zdroj: | Biochemical and Biophysical Research Communications. 308:713-718 |
| ISSN: | 0006-291X |
| DOI: | 10.1016/s0006-291x(03)01462-1 |
| Popis: | The SV40t polyadenylation and splicing signals of the pAEC plasmid vectors were replaced by synthetic intron and synthetic rabbit beta globin-based termination/polyadenylation sequences, and 5, 10, and 20 copies of the 5'-AACGTT-3' CpG motif were inserted. Balb/c mice were immunized by intramuscular injection of 200 microg of each plasmid, coding for the HIV-1 multiepitope TAB9, under the control of the human cytomegalovirus promoter. After three doses of DNA, a fourth boost with plasmid DNA or a TAB9-expressing recombinant fowlpox virus rFPTAB9LZ was administered. ELISA and ELISPOT assays were conducted for antibody and IFN-gamma-secreting cell-mediated responses' evaluation against the whole TAB9 and the TAB9's IIIB V3 peptide, respectively. Serum IgG antibodies were not detected. Effector IFN-gamma-secreting responses were only detected on the animals receiving the new set of DNA constructs, alone or in combination with a recombinant virus boost, with or without in vitro re-stimulation. The response was dependent on the new transcriptional unit and influenced by the number of CpG motifs. We showed that plasmid backbone optimization based on these two factors could enhance the response against a multiepitope-based DNA vaccine. A new family of plasmid vectors is also available for evaluation with desired antigens. |
| Databáze: | OpenAIRE |
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