title: A multicistronic DNA vaccine induces significant protection against tuberculosis in mice and offers flexibility in the expressed antigen repertoire.
abstract: Concerns about the safety and efficacy of Mycobacterium bovis bacillus Calmette-Guérin (BCG) emphasize the need for alternative tuberculosis (TB) vaccines. DNA vaccines are interesting candidates but are limited by the restricted antigen repertoire that they express. Traditional polycistronic vectors are large and have imbalanced expression. Recent advances in molecular genetics and cellular immunology have paved the way toward the rational design of an efficacious vaccine. We exploited self-cleaving peptide 2A from the foot-and-mouth disease virus, because of its small size and high cleavage activity, to generate an efficient TB DNA vaccine (V-2A). V-2A expresses three mycobacterial antigens, Rv3407, Ag85A, and HspX, in a single open reading frame joined by the 2A sequences, which lead to the segmentation of the long translated polypeptide into individual proteins by posttranslational modification. Our in vitro measurements revealed no differences at the transcriptional or translational level between V-2A and the monocistronic expression of the individual antigens. Mice vaccinated with V-2A developed antigen-specific cellular and humoral responses against all three antigens, imparting protection against Mycobacterium tuberculosis aerosol challenge equivalent to that imparted by BCG. These results have important implications for the rational design and development of efficacious recombinant subunit vaccines.
authors: Department of Immunology, Max Planck Institute for Infection Biology, Charitéplatz 1, Berlin 10117, Germany.
authors: Fayaz-Ahmad
authors: FA
authors: Mir
authors: Stefan H E
authors: SH
authors: Kaufmann
authors: Ali Nasser
authors: AN
authors: Eddine
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics; D001500:BCG Vaccine / Q000494:pharmacology; D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D005260:Female; D029544:Foot-and-Mouth Disease Virus / Q000235:genetics; D005796:Genes; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000276:immunology; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology; D014376:Tuberculosis / Q000276:immunology / Q000517:prevention & control*; D032581:Tuberculosis Vaccines / Q000235:genetics / Q000494:pharmacology*; D019444:Vaccines, DNA / Q000235:genetics / Q000494:pharmacology; D014764:Viral Proteins / Q000235:genetics
chemical_list: D000942:Antigens, Bacterial; D001500:BCG Vaccine; D017931:DNA Primers; D004269:DNA, Bacterial; D011994:Recombinant Proteins; D032581:Tuberculosis Vaccines; D019444:Vaccines, DNA; D014764:Viral Proteins; C057421:virus protein 2A