Transfer of a pertussis toxin expression locus to isogenicbvg-positive andbvg-negative strains ofBordetella bronchisepticausing anin vivotechnique

Smith, Adam M; Walker, Mark J.

doi:10.1006/mpat.1996.0025

Cited by 5 publications

(5 citation statements)

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“…A 2.2 kb kanamycin resistance cassette was excised from pHP45ΩKm with Eco RI, Klenow treated and ligated into the now unique Sal I site of pJMC10, which had also been Klenow treated, to create pJMC13. After conjugation [29] between S17‐1λ pir containing pJMC13 and B. bronchiseptica K8744/1b, co‐integrates were selected for SS‐X agar plates containing aamix, Km and Cp. Aro − mutants were identified by their reduced, but not abolished ability to grow on SS‐X medium lacking aamix.…”

Section: Methodsmentioning

confidence: 99%

An aromatic amino acid auxotrophic mutant ofBordetella bronchisepticais attenuated and immunogenic in a mouse model of infection

McArthur

West

Cole

et al. 2003

FEMS Microbiology Letters

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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. bronchisepticaspecific 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.

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Section: Methodsmentioning

confidence: 99%

An aromatic amino acid auxotrophic mutant ofBordetella bronchisepticais attenuated and immunogenic in a mouse model of infection

McArthur

West

Cole

et al. 2003

FEMS Microbiology Letters

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“…The method by which the mini-transposon was transferred from BB7866pgm to the homologous location on the wild-type chromosome (BB7865pgm) was that described by Smith and Walker (44). E. coli Q358(pR715::Tn813) was mated with BB7866pgm.…”

Section: Methodsmentioning

confidence: 99%

“…An open reading frame located downstream of the pgi gene from B. pertussis has been identified as encoding a possible galactosyl transferase; however, this gene is not included within the pgm operon. The identical gene was mutated in BB7865, the parental strain of BB7866, by way of an in vivo chromosome transfer technique (44). This allowed for the effects of such a mutation to be observed in a bvg-positive background.…”

Section: Identification Of a Novel Lps Genetic Locus In B Bronchisepmentioning

confidence: 99%

Role of Phosphoglucomutase of Bordetella bronchiseptica in Lipopolysaccharide Biosynthesis and Virulence

et al. 2000

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The phosphoglucomutase (PGM)-encoding gene of Bordetella bronchiseptica is required for lipopolysaccharide (LPS) biosynthesis. An insertion mutant of the wild-type B. bronchiseptica strain BB7865 which disrupted LPS biosynthesis was created and characterized (BB7865pgm). Genetic analysis of the mutated gene showed it shares high identity with PGM genes of various bacterial species and forms part of an operon which also encompasses the gene encoding phosphoglucose isomerase. Functional assays for PGM revealed that enzyme activity is expressed in both bvg-positive and bvg-negative strains of B. bronchiseptica and is substantially reduced in BB7865pgm. Complementation of the mutated PGM gene with that from BB7865 restored the wildtype condition for all phenotypes tested. The ability of the mutant BB7865pgm to survive within J774.A1 cells was significantly reduced at 2 h (40% reduction) and 24 h (56% reduction) postinfection. BB7865pgm was also significantly attenuated in its ability to survive in vivo following intranasal infection of mice, being effectively cleared from the lungs within 4 days, whereas the wild-type strain persisted at least 35 days. The activities of superoxide dismutase, urease, and acid phosphatase were unaffected in the PGM-deficient strain. In contrast, the inability to produce wild-type LPS resulted in a reduced bacterial resistance to oxidative stress and a higher susceptibility to the antimicrobial peptide cecropin P.

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“…Unfortunately, the toxin was not secreted to the growth medium, making this strain less than suitable as a vaccine expression system. Further study demonstrated that neither virulent nor avirulent strains of B. bronchiseptica were capable of PT secretion [171].…”

Section: Virulence Factors Of B Pertussismentioning

confidence: 93%

“…The ptx/ptl promoter (filled arrow; P wt ) is also depicted. Modified from Smith and Walker [171]; Farizo et al [173].…”

Section: Virulence Factors Of B Pertussismentioning

confidence: 99%

The virulence factors ofBordetella pertussis: a matter of control

2001

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Bordetella pertussis is the causative agent of whooping cough, a contagious childhood respiratory disease. Increasing public concern over the safety of whole-cell vaccines led to decreased immunisation rates and a subsequent increase in the incidence of the disease. Research into the development of safer, more efficacious, less reactogenic vaccine preparations was concentrated on the production and purification of detoxified B. pertussis virulence factors. These virulence factors include adhesins such as filamentous haemagglutinin, fimbriae and pertactin, which allow B. pertussis to bind to ciliated epithelial cells in the upper respiratory tract. Once attachment is initiated, toxins produced by the bacterium enable colonisation to proceed by interfering with host clearance mechanisms. B. pertussis co-ordinately regulates the expression of virulence factors via the Bordetella virulence gene (bvg) locus, which encodes a response regulator responsible for signal-mediated activation and repression. This strict regulation mechanism allows the bacterium to express different gene subsets in different environmental niches within the host, according to the stage of disease progression.

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Transfer of a pertussis toxin expression locus to isogenicbvg-positive andbvg-negative strains ofBordetella bronchisepticausing anin vivotechnique

Cited by 5 publications

References 0 publications

An aromatic amino acid auxotrophic mutant ofBordetella bronchisepticais attenuated and immunogenic in a mouse model of infection

An aromatic amino acid auxotrophic mutant ofBordetella bronchisepticais attenuated and immunogenic in a mouse model of infection

Role of Phosphoglucomutase of Bordetella bronchiseptica in Lipopolysaccharide Biosynthesis and Virulence

The virulence factors ofBordetella pertussis: a matter of control

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