title: In vivo induction of CTL responses by recombinant adenylate cyclase of Bordetella pertussis carrying multiple copies of a viral CD8(+) T-cell epitope.
abstract: Bordetella pertussis adenylate cyclase toxin (ACT) is one of the few known protein toxins penetrating directly into the cytosol of target cells across their cytoplasmic membrane without the need for endocytosis. This capacity of ACT was recently exploited for in vivo delivery of single viral CD8(+) T-epitopes into MHC class I-presenting cells and induction of protective antiviral cytotoxic T-cell (CTL) responses. Here, we have explored the potential of the cell-invasive adenylate cyclase domain of the toxin to deliver larger antigens by evaluating the epitope-specific CTL responses induced by constructs bearing one to four copies of the CD8(+) T-epitope from the nucleoprotein of the lymphocytic choriomeningitis virus. The increase in the number of copies of the epitope was accompanied by a moderate decrease of the specific cell invasiveness of the ACT protein and did not lead to further enhancement of the level of induced epitope-specific CTL cells in mice, as compared to ACT with a single copy of the epitope. These results demonstrate the capacity of ACT to deliver larger heterologous antigens comprising several epitopes for antigenic presentation in vivo.
authors.affiliation: Cell and Molecular Microbiology Division, Institute of Microbiology of the Czech Academy of Sciences, Videnská 1083, CZ-142 20, Prague, Czech Republic. sebo@biomed.cas.cz
authors.forename: P
authors.initials: P
authors.lastname: Sebo
authors.forename: Z
authors.initials: Z
authors.lastname: Moukrim
authors.forename: M
authors.initials: M
authors.lastname: Kalhous
authors.forename: N
authors.initials: N
authors.lastname: Schaft
authors.forename: G
authors.initials: G
authors.lastname: Dadaglio
authors.forename: V
authors.initials: V
authors.lastname: Sheshko
authors.forename: C
authors.initials: C
authors.lastname: Fayolle
authors.forename: C
authors.initials: C
authors.lastname: Leclerc
mesh_terms: D037361:Adenylate Cyclase Toxin; D000818:Animals; D001886:Bordetella pertussis / Q000737:chemistry*; D016827:CD8 Antigens / Q000235:genetics*; D003602:Cytotoxicity, Immunologic; D000939:Epitopes; D004926:Escherichia coli / Q000378:metabolism; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C / Q000276:immunology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology*; D014611:Vaccination; D010566:Virulence Factors, Bordetella / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*
chemical_list: D037361:Adenylate Cyclase Toxin; D016827:CD8 Antigens; D000939:Epitopes; D011994:Recombinant Proteins; D010566:Virulence Factors, Bordetella