Construction of an unmarked recombinant BCG expressing a pertussis antigen by auxotrophic complementation: protection against Bordetella pertussis challenge in neonates.
Mycobacterium bovis BCG has long been investigated as a candidate for heterologous antigen presentation. We have previously described an rBCG-Pertussis that confers protection against challenge with Bordetella pertussis in neonate and adult mice. In order to obtain stable expression in vivo, we constructed an unmarked BCG lysine auxotrophic and a complementation vector containing the lysine and the genetically detoxified S1 pertussis toxin genes, both under control of the same promoter. Complemented BCG-Delta lysine growth and expression of the pertussis antigen were stable, without the use of an antibiotic marker. Our results show that the complemented rBCG-Delta lysA-S1PT-lysA(+)(kan(-)), which is now suitable to be evaluated in clinical trials, maintains similar characteristics of the original rBCG-pNL71S1PT strain, such as the antigen expression level, cellular immune response and protection against the same model challenge in neonatal-immunized mice.
Nascimento
Ivan P
IP
Centro de Biotecnologia, Instituto Butantan, Av. Vital Brasil 1500, 05503-900 São Paulo, SP, Brazil.
Dias
Waldely O
WO
Quintilio
Wagner
W
Hsu
Tsungda
T
Jacobs
William R
WR
Leite
Luciana C C
LC
D000818:Animals; D000831:Animals, Newborn; D000942:Antigens, Bacterial / Q000276:immunology*; D001886:Bordetella pertussis / Q000276:immunology; D003001:Cloning, Molecular; D005822:Genetic Vectors; D007111:Immunity, Cellular; D007371:Interferon-gamma / Q000276:immunology; D051379:Mice; D009163:Mycobacterium bovis / Q000276:immunology*; D037342:Pertussis Toxin / Q000276:immunology*; D010567:Pertussis Vaccine / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000276:immunology; D014161:Transduction, Genetic; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology; D014917:Whooping Cough / Q000276:immunology / Q000517:prevention & control*
D000942:Antigens, Bacterial; D010567:Pertussis Vaccine; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma; D037342:Pertussis Toxin