title: An aromatic amino acid auxotrophic mutant of Bordetella bronchiseptica is attenuated and immunogenic in a mouse model of infection.
abstract: We have constructed an aromatic amino acid auxotrophic mutant of Bordetella bronchiseptica, harbouring mutations in aroA and trpE to investigate the use of such a strain as a live-attenuated vaccine. B. bronchiseptica aroA trpE was unable to grow in minimal medium without aromatic supplementation. Compared to the parental wild-type strain, the mutant displayed significantly reduced abilities to invade and survive within the mouse macrophage-like cell line J774A.1 in vitro and in the murine respiratory tract following experimental intranasal infection. Mice vaccinated with B. bronchiseptica aroA trpE displayed significant dose-dependent increases in B. bronchiseptica-specific antibody responses, and exhibited increases in the number of B. bronchiseptica-reactive spleen cells in lymphoproliferation assays. Immunised animals were protected against lung colonisation after challenge with the wild-type parental strain. With such a broad host range displayed by B. bronchiseptica, the attenuated strain constructed in this study may not only be used for the prevention of B. bronchiseptica-associated disease, but also for the potential delivery of heterologous antigen.
authors.affiliation: Department of Biological Sciences, University of Wollongong, Wollongong, NSW 2522, Australia.
authors.forename: Jason D
authors.initials: JD
authors.lastname: McArthur
authors.forename: Nicholas P
authors.initials: NP
authors.lastname: West
authors.forename: Jason N
authors.initials: JN
authors.lastname: Cole
authors.forename: Heidrun
authors.initials: H
authors.lastname: Jungnitz
authors.forename: Carlos A
authors.initials: CA
authors.lastname: Guzmán
authors.forename: James
authors.initials: J
authors.lastname: Chin
authors.forename: Philip R
authors.initials: PR
authors.lastname: Lehrbach
authors.forename: Steven P
authors.initials: SP
authors.lastname: Djordjevic
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Walker
mesh_terms: D051229:3-Phosphoshikimate 1-Carboxyvinyltransferase; D019883:Alkyl and Aryl Transferases / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D024322:Amino Acids, Aromatic / Q000378:metabolism*; D000818:Animals; D000878:Anthranilate Synthase / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D000907:Antibodies, Bacterial / Q000097:blood; D001428:Bacterial Vaccines / Q000008:administration & dosage / Q000276:immunology*; D001885:Bordetella Infections / Q000382:microbiology / Q000517:prevention & control*; D016950:Bordetella bronchiseptica / Q000235:genetics / Q000254:growth & development / Q000276:immunology* / Q000472:pathogenicity; D002460:Cell Line; D004195:Disease Models, Animal; D005260:Female; D008213:Lymphocyte Activation; D008264:Macrophages / Q000382:microbiology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009154:Mutation; D009876:Operon; D017422:Sequence Analysis, DNA; D014611:Vaccination; D014613:Vaccines, Attenuated / Q000008:administration & dosage / Q000276:immunology*
chemical_list: D024322:Amino Acids, Aromatic; D000907:Antibodies, Bacterial; D001428:Bacterial Vaccines; D014613:Vaccines, Attenuated; D019883:Alkyl and Aryl Transferases; D051229:3-Phosphoshikimate 1-Carboxyvinyltransferase; D000878:Anthranilate Synthase