Aerosol infection of mice with Bordetella pertussis

Sato, Yuji; Izumiya, K; Sato, Hiroko; Cowell, J L; Manclark, C R

doi:10.1128/iai.29.1.261-266.1980

Cited by 94 publications

(80 citation statements)

References 19 publications

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“…A 21 h culture of B. pertussis grown on BG agar with or without 50 mM MgSO 4 was suspended in sterile PBS or PBS + MgSO 4 , respectively, at a concentration of approximately 2 ¥ 10 9 cfu ml -1 . Each different strain was administered to 17 day old BALB/cAnNCr mice (Animal Production Program, Division of Cancer Treatment, National Cancer Institute, Frederick, Md) in a separate aerosol challenge for 30 min as described previously (Sato et al, 1980;Shahin et al, 1990). Mice were removed from the chamber 1 h after termination of the aerosol challenge.…”

Section: Mouse Aerosol Challengementioning

confidence: 99%

Demonstration of differential virulence gene promoter activation in vivo in Bordetella pertussis using RIVET

Veal-Carr

Stibitz

2004

Molecular Microbiology

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SummaryBordetella pertussis , the etiologic agent of whooping cough, causes disease by employing an array of virulence factors controlled by the BvgA-BvgS twocomponent signal transduction system. Regulation by this system has been extensively characterized in vitro , where bvg -activated genes are repressed in a process known as phenotypic modulation. Differential regulation of these genes by the response regulator BvgA results in promoters that are activated early, middle, or late after being released from modulation. However, the in vivo environmental signal and regulation pattern has not been described. In order to investigate BvgAS-mediated regulation of B. pertussis virulence factors in vivo using the mouse aerosol challenge model, we have adapted the recombinasebased in vivo technology (RIVET) system for use in B. pertussis . We have demonstrated that these strains show resolution during in vitro growth under nonmodulating conditions. In addition, we have demonstrated that modulating strains by growth on media containing MgSO 4 does not affect virulence in the mouse aerosol challenge model. We have therefore used the RIVET system to reveal the time-course of gene expression in vivo for selected B. pertussis virulence factors ( cya , fha , prn and ptx ). Our data indicate that this method can be effectively used to monitor and compare in vivo and in vitro gene expression in B. pertussis , and that temporal regulation patterns previously observed in vitro are mirrored in vivo .

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Section: Mouse Aerosol Challengementioning

confidence: 99%

Demonstration of differential virulence gene promoter activation in vivo in Bordetella pertussis using RIVET

Veal-Carr

Stibitz

2004

Molecular Microbiology

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“…Respiratory infection of mice was performed by aerosol challenge using a modification of the method described by Sato et al 19 Bacteria from a 48-hr culture were resuspended in physiological saline containing 1% casein at a concentration of 2 2 10 10 colonyforming units (CFU)/ml. The challenge inoculum was administered to the mice for 12 min using a nebulizer in a sealed container within a class 3 exhaust protected cabinet.…”

Section: Bordetella Pertussis Challengementioning

confidence: 99%

Th1/Th2 cell dichotomy in acquired immunity to Bordetella pertussis: variables in the in vivo priming and in vitro cytokine detection techniques affect the classification of T‐cell subsets as Th1, Th2 or Th0

et al. 1996

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SUMMARYIn studies of the mechanism of immunity to Bordetella pertussis in a murine respiratory infection model, we have previously demonstrated that natural infection of immunization with a whole cell vaccine induces a potent protective immune response, which is mediated by T-helper type-1 (Th1) cells. In contrast an acellular vaccine generates Th2 cells and is associated with delayed bacterial clearance following respiratory challenge. In the present study we have investigated the apparent Th1/Th2 cell dichotomy in acquired immunity and have examined the factors that affect their induction or detection. The cytokine profiles of B. pertussis-specific T cells in immune animals were determined using antigenstimulated ex vivo spleen cells or CD4T-cell lines and clones established in the presence of interleukin-2 (IL-2) or IL-4. Antigen-specific T cells derived from mice immunized with the acellular vaccine were almost exclusively of the Th2 cell type. In contrast, T-cell lines and clones established following respiratory infection or immunization with the whole cell vaccine were predominantly of the Th1 type. However, a proportion of T cells from convalescent mice, especially when cultured in the presence of IL-4, secreted IL-4 and IL-5 with or without detectable IL-2 and interferon-g (IFN-g), suggesting that Th0 or Th2 cells were also primed during natural infection in vivo. Furthermore, when mice were assessed 6 months after infection, spleen cells produced significant levels of IL-4 and IL-5, which were not evident at 6 weeks. The route of immunization and the genetic background of the mice were also found to influence the preferential priming of Th1 cells, and this was directly related to the level of protection against respiratory or intracerebral (i.c.) challenge. Our findings underline the critical role of CD4 Th1 cells in immunity to B. pertussis, but also demonstrate that a number of factors in the in vivo priming and in vitro restimulation can skew the apparent dominance of one Th cell type over another.

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“…The intracerebral challenge model, which is currently employed in vaccine testing, poses several problems since not all virulent strains of B. pertussis are active in this assay (Standfast, 1958). New approaches are being studied (Sato et al, 1980) and these may provide better models for the disease, enabling us to describe virulence more precisely in genetic terms. That variation is detrimental to pathogenicity via the intracerebral route was suggested by Standfast (1958) and supported by our inability to challenge mice intracerebrally with the # 165 strain (unpublished data).…”

Section: Discussionmentioning

confidence: 99%

Spontaneous phase variation in Bordetella pertussis is a multistep non-random process.

Goldman¹,

Hanski²,

Fish³

1984

The EMBO Journal

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Pathogenic strains of Bordetella pertussis undergo spontaneous phase variation and become non‐pathogenic upon culturing in vitro. Spontaneous variants of the Tohama and #165 pathogenic strains of B. pertussis were selected by their ability to grow on synthetic and semi‐synthetic solid media. The frequency of these variants was between 10(‐6) and 10(‐7). About 250 variant strains were screened for the presence of virulence‐associated traits, such as production of hemolysin, pertussis toxin and filamentous hemagglutinin (FHA). Only four different combinations of the traits were found: 7‐11% of the variants displayed all traits, 17% of the variants carried the toxin and FHA, 5‐11% carried FHA only and 66% were devoid of all virulence traits. The strains which had at least one virulence trait also demonstrated some adenylate cyclase activity. The disappearance of hemolysin quantitatively affected the other traits. These results suggest that phase variation in B. pertussis is a non‐random process, involving multistep disappearance of virulence factors in the following order: hemolysin, pertussis toxin and FHA. In contrast, all 300 variants of strain #18323 of B. pertussis, which were able to grow on the selective solid media, carried all the virulence traits. This is in accordance with the strain's unique intracerebral growth capability.

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Aerosol infection of mice with Bordetella pertussis

Cited by 94 publications

References 19 publications

Demonstration of differential virulence gene promoter activation in vivo in Bordetella pertussis using RIVET

Demonstration of differential virulence gene promoter activation in vivo in Bordetella pertussis using RIVET

Th1/Th2 cell dichotomy in acquired immunity to Bordetella pertussis: variables in the in vivo priming and in vitro cytokine detection techniques affect the classification of T‐cell subsets as Th1, Th2 or Th0

Spontaneous phase variation in Bordetella pertussis is a multistep non-random process.

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