title: Immunogenicity and protective efficacy of a tuberculosis DNA vaccine co-expressing pro-apoptotic caspase-3.
abstract: DNA vaccination is a potent means for inducing strong cell-mediated immune responses and protective immunity against viral, bacterial and parasite pathogens in rodents. In an attempt to increase cross-presentation through apoptosis, the DNA-encoding caspase-2 prodomain followed by wild-type or catalytically inactive mutated caspase-3 was inserted into a plasmid encoding the 32 kDa mycolyl transferase (Ag85A) from Mycobacterium tuberculosis. Transient transfection showed that the mutated caspase induced slow apoptosis, normal protein expression and NF-kappaB activation while wild-type caspase induced rapid apoptosis, lower protein expression and no NF-kappaB activation. Ag85A specific antibody production was increased by co-expressing the mutated and decreased by co-expressing the wild-type caspase. Vaccination with pro-apoptotic plasmids triggered more Ag85A specific IFN-gamma producing spleen cells, and more efficient IL-2 and IFN-gamma producing memory cells in spleen and lungs after M. tuberculosis challenge. Compared to DNA-encoding secreted Ag85A, vaccination with DNA co-expressing wild-type caspase increased protection after infection with M. tuberculosis, while vaccination with plasmid co-expressing mutated caspase was not protective, possibly due to the stimulation of IL-6, IL-10 and IL-17A production.
authors.affiliation: Department of Molecular and Cellular Interactions, Vlaams Instituut voor Biotechnologie, Brussels, Belgium.
authors.forename: Tatiana
authors.initials: T
authors.lastname: Gartner
authors.forename: Marta
authors.initials: M
authors.lastname: Romano
authors.forename: Vanessa
authors.initials: V
authors.lastname: Suin
authors.forename: Michaël
authors.initials: M
authors.lastname: Kalai
authors.forename: Hannelie
authors.initials: H
authors.lastname: Korf
authors.forename: Patrick
authors.initials: P
authors.lastname: De Baetselier
authors.forename: Kris
authors.initials: K
authors.lastname: Huygen
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000235:genetics* / Q000276:immunology*; D015153:Blotting, Western; D053143:Caspase 2 / Q000235:genetics / Q000276:immunology; D053148:Caspase 3 / Q000096:biosynthesis*; D002460:Cell Line; D004269:DNA, Bacterial / Q000096:biosynthesis / Q000276:immunology; D004789:Enzyme Activation / Q000502:physiology; D005434:Flow Cytometry; D007111:Immunity, Cellular; D007371:Interferon-gamma / Q000096:biosynthesis / Q000235:genetics; D007376:Interleukin-2 / Q000096:biosynthesis; D008168:Lung / Q000166:cytology / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D050506:Mutant Chimeric Proteins / Q000276:immunology; D009169:Mycobacterium tuberculosis / Q000254:growth & development / Q000276:immunology; D016328:NF-kappa B / Q000235:genetics / Q000276:immunology; D010957:Plasmids / Q000235:genetics / Q000276:immunology; D013154:Spleen / Q000166:cytology / Q000276:immunology; D013534:Survival; D014162:Transfection; D032581:Tuberculosis Vaccines / Q000235:genetics* / Q000627:therapeutic use*; D019444:Vaccines, DNA / Q000627:therapeutic use
chemical_list: D004269:DNA, Bacterial; D007376:Interleukin-2; D050506:Mutant Chimeric Proteins; D016328:NF-kappa B; D032581:Tuberculosis Vaccines; D019444:Vaccines, DNA; D007371:Interferon-gamma; D053143:Caspase 2; D053148:Caspase 3