Effects of Bordetella pertussis infection on human respiratory epithelium in vivo and in vitro.

ABSTRACT Pertussis is an acute respiratory disease caused by the bacterium Bordetella pertussis, for which humans are the only known reservoir. During infection, B. pertussis releases several toxins, including pertussis toxin (PT) and adenylate cyclase toxin (ACT), which have both been shown to play roles in promoting bacterial growth during early infection in a mouse model. Furthermore, in vitro and in vivo studies suggest that PT and ACT affect neutrophil chemotaxis and/or function, thereby altering the innate immune response. In this study we depleted animals of neutrophils to investigate whether neutrophils play a protective role during B. pertussis infection in mice. In addition, by infection with toxin-deficient strains, we investigated whether neutrophils are the main targets for PT and/or ACT activity in promoting bacterial growth. Surprisingly, we found no role for neutrophils during B. pertussis infection in naïve mice. However, in previously infected (immune) mice or in mice receiving immune serum, we observed a significant role for neutrophils during infection. Furthermore, in this immune mouse model our evidence indicates that neutrophils appear to be the main target cells for ACT, but not for PT.

Download Full-text

Strain-Dependent Role of BrkA during Bordetella pertussis Infection of the Murine Respiratory Tract

Infection and Immunity ◽

10.1128/iai.72.10.5919-5924.2004 ◽

2004 ◽

Vol 72 (10) ◽

pp. 5919-5924 ◽

Cited By ~ 20

Author(s):

Kelly D. Elder ◽

Eric T. Harvill

Keyword(s):

Respiratory Tract ◽

Bordetella Pertussis ◽

Whooping Cough ◽

Single Gene ◽

Wild Type ◽

Pertussis Infection ◽

Redundant Functions

ABSTRACT Bordetella pertussis, the causative agent of whooping cough, expresses many virulence factors believed to be involved in infection and disease progression. While these factors as a group are required for infection, deletion of individual virulence factor genes generally has limited effects on the ability of B. pertussis to efficiently infect the respiratory tract of mice, suggesting they may perform noncritical or redundant functions. We have recently observed that a B. pertussis strain, putatively with a mutation of a single gene, brkA, results in a severe defect in vivo. Although BrkA has been shown to be required for B. pertussis to resist complement-mediated killing in vitro, the relevance of these findings to the in vivo role of BrkA during infection has not been examined. Transducing this mutation into multiple wild-type B. pertussis strains allowed us to confirm the in vitro phenotype of reduced resistance to serum complement. All ΔbrkA mutants were increased in their sensitivity to complement in vitro, both in the presence and absence of antibodies. However, these strains differed substantially in their phenotypes in vivo. ΔbrkA mutants of recent clinical isolates were indistinguishable from wild-type strains in their efficient infection of respiratory organs, suggesting that the function of BrkA in these strains is noncritical or redundant. In contrast, multiple ΔbrkA strains derived from Tohama I were severely defective during the first week postinoculation compared to their wild-type parent. This defect was present even in complement-deficient mice, revealing a complement-independent phenotype for the ΔbrkA mutant in respiratory tract infection.

Download Full-text

The intracerebral infection of mice with Bordetella pertussis

Journal of Hygiene ◽

10.1017/s0022172400038869 ◽

1961 ◽

Vol 59 (2) ◽

pp. 205-216 ◽

Cited By ~ 11

Author(s):

Jean M. Dolby ◽

A. F. B. Standfast

Keyword(s):

Bordetella Pertussis ◽

Viable Count ◽

Lethal Level ◽

Virulent Strains ◽

Single Organism ◽

Striking Change ◽

Circulating Antibodies ◽

Intracerebral Infection

The growth of virulent strains of Bordetella pertussis in the brains of mice was studied by carrying out viable counts on mice killed at various times during the infection. The results suggested that this system conformed to the general model which postulates that the organisms causing death multiply in vivo at a rate which is constant for all doses and that death is certain to occur when the number of organisms reaches a certain constant figure.Perhaps the most important factor in this route of infection is the lodgement of the parasite in the host, for if this is accomplished a single organism grows until the lethal level is reached. There is no sublethal infection.In actively and passively protected mice, the growth of the organism is approximately the same as in unprotected controls for the first 4–5 days. At this time there is a striking change in protected animals and the viable count falls rapidly and progressively and the animals survive. At the same time the blood-brain barrier becomes permeable and circulating antibodies diffuse into the brain. In vitro, specific antisera plus complement are highly bactericidal.

Download Full-text

Functional Differences in Pertussis Toxins from Bordetella pertussis Clinical Isolates as Determined by in vitro and in vivo Assays

Journal of Clinical & Experimental Pathology ◽

10.4172/2161-0681.1000243 ◽

2015 ◽

Vol 05 (05) ◽

Author(s):

Catpagavalli Asokanathan ◽

Chun-Ting Yuen

Keyword(s):

Bordetella Pertussis ◽

Clinical Isolates ◽

In Vivo Assays ◽

Functional Differences

Download Full-text

The effect of the antigen which elicits the bactericidal antibody and of the mouse-protective antiǵen on the growth of Bordetella pertussis in the mouse brain

Journal of Hygiene ◽

10.1017/s0022172400046738 ◽

1975 ◽

Vol 74 (1) ◽

pp. 71-83 ◽

Cited By ~ 3

Author(s):

Jean M. Dolby ◽

J. P. Ackers ◽

D. E. Dolby

Keyword(s):

Bordetella Pertussis ◽

Protective Antigen ◽

Passive Immunity ◽

Cell Vaccine ◽

Potency Assay ◽

Whole Cell ◽

Protective Antibody ◽

Bactericidal Antibody

SUMMARYThe effect of antigens of Bordetella pertussis and their antibodies on brain infections by B. pertussis in mice are suppression of an infection immediately, so that the initial 90 % loss due to leakage from the brain is maintained or the numbers of bacteria are reduced even further, sometimes with complete sterilization particularly after a small lethal challenge of 10 LD 50 (mechanism 1), and a delayed antibacterial activity in vivo which does not begin until 3 days after challenge (mechanism 2). The first, immediate reaction is over in 2–3 days; the second is maintained from 3–4 days onwards, and results in elimination of the bacteria and protection of mice.The parts played in vivo in overcoming infection in these two ways by two antigens and their respective antibodies have been investigated. These antigens are a lipopolysaccharide capable of eliciting an antibody which is bactericidal in vitro in the presence of complement called the ‘bactericidal antigen’, and the mouse protective antigen.Considering first passive immunity, bactericidal antibody elicited by isolated antigen, and of high titre in vitro, is only very weakly active by mechanism (1) in vivo. Brains are seldom sterilized and mice not therefore protected. Antisera to whole cell vaccines whether they contain the ‘bactericidal antigen’ or not, or the protective antigen or not can more easily reduce infections by mechanism (1), eliminating small lethal challenges in some mice which are protected. A passive, intracerebrally protective antibody (PIPA) different from other known antibodies, has been postulated to account for this. Antisera to whole cell vaccine which is protective as denned in the potency assay, can, in additon to this, protect mice by mechanism (2) not only against 10 LD 50 but also 100 LD 50 challenge, and is the only antibody which can do this.These antibodies have been investigated by injecting them with the challenging organisms. The antibody effects described above are given by antisera stimulated by several injections and also by the concentrated serum immunoglobulins of once vaccinated mice. The antibody, which is bactericidal in vitro only, is in the 7 S globulin fraction of the serum of once vaccinated mice. The protective antibody capable of overcoming small and large challenges is in the 19 S and 11 S globulins. The antibody, PIPA, protecting against small lethal challenges only is in the fraction A2 containing mainly 11 S globulin.In active immunization experiments the suppression of infection which immediately follows intracerebral vaccination, but which only lasts 2–3 days (mechanism 1), is not dependent on either ‘bactericidal’ or protective antigens but on a component present in all our whole cell vaccines. Vaccines which also had protective antigen eliminated the remaining infection at 4–6 days after challenge by mechanism (2).As in passive immunity, only the protective antigen can completely overcome 100 LD 50. Suppression of a small, lethal, intracerebral infection given 14 days after intraperitoneal vaccination by mechanism (1) may however be correlated with protective antigen.

Download Full-text

In-vitro and in-vivo analysis of the production of the Bordetella type three secretion system effector A in Bordetella pertussis, Bordetella parapertussis and Bordetella bronchiseptica

Microbes and Infection ◽

10.1016/j.micinf.2013.02.006 ◽

2013 ◽

Vol 15 (5) ◽

pp. 399-408 ◽

Cited By ~ 14

Author(s):

Nicolas Hegerle ◽

Lamya Rayat ◽

Grégory Dore ◽

Nora Zidane ◽

Hugues Bedouelle ◽

...

Keyword(s):

Bordetella Pertussis ◽

Secretion System ◽

Bordetella Bronchiseptica ◽

Bordetella Parapertussis ◽

Type Three Secretion System ◽

In Vivo Analysis ◽

Type Three Secretion

Download Full-text

Analysis of bvgR Expression in Bordetella pertussis

Journal of Bacteriology ◽

10.1128/jb.185.23.6902-6912.2003 ◽

2003 ◽

Vol 185 (23) ◽

pp. 6902-6912 ◽

Cited By ~ 27

Author(s):

Tod J. Merkel ◽

Philip E. Boucher ◽

Scott Stibitz ◽

Vanessa K. Grippe

Keyword(s):

Bordetella Pertussis ◽

Transcriptional Activation ◽

Northern Blot Analysis ◽

Whooping Cough ◽

Transmembrane Protein ◽

In Vitro Transcription ◽

Environmental Signals ◽

Transcription System

ABSTRACT Bordetella pertussis, the causative agent of whooping cough, produces a wide array of factors that are associated with its ability to cause disease. The expression and regulation of these virulence factors are dependent upon the bvg locus, which encodes three proteins: BvgA, a 23-kDa cytoplasmic protein; BvgS, a 135-kDa transmembrane protein; and BvgR, a 32-kDa protein. It is hypothesized that BvgS responds to environmental signals and interacts with BvgA, a transcriptional regulator, which upon modification by BvgS binds to specific promoters and activates transcription. An additional class of genes is repressed by the products of the bvg locus. The repression of these genes is dependent upon the third gene, bvgR. Expression of bvgR is dependent upon the function of BvgA and BvgS. This led to the hypothesis that the binding of phosphorylated BvgA to the bvgR promoter activates the expression of bvgR. We undertook an analysis of the transcriptional activation of bvgR expression. We identified the bvgR transcript by Northern blot analysis and identified the start site of transcription by primer extension. We determined that transcriptional activation of the bvgR promoter in an in vitro transcription system requires the addition of phosphorylated BvgA. Additionally, we have identified cis-acting regions that are required for BvgA activation of the bvgR promoter by in vitro footprinting and in vivo deletion and linker scanning analyses. A model of BvgA binding to the bvgR promoter is presented.

Download Full-text

Role of Adenylate Cyclase-Hemolysin in Alveolar Macrophage Apoptosis during Bordetella pertussis Infection In Vivo

Infection and Immunity ◽

10.1128/iai.66.4.1718-1725.1998 ◽

1998 ◽

Vol 66 (4) ◽

pp. 1718-1725 ◽

Cited By ~ 97

Author(s):

Pascale Gueirard ◽

Anne Druilhe ◽

Marina Pretolani ◽

Nicole Guiso

Keyword(s):

Adenylate Cyclase ◽

Bronchoalveolar Lavage ◽

Bordetella Pertussis ◽

Alveolar Macrophages ◽

Apoptotic Cells ◽

Neutrophil Accumulation ◽

Pertussis Infection ◽

Intranasal Infection

ABSTRACT Bordetella pertussis induces in vitro apoptosis of murine alveolar macrophages by a mechanism that is dependent on expression of bacterial adenylate cyclase-hemolysin. Using a murine respiratory model, we found in this study that intranasal infection with a parental B. pertussis strain, but not with an isogenic variant deficient in the expression of all toxins and adhesins, induced a marked neutrophil accumulation in the bronchoalveolar lavage fluid and an early decrease in macrophage numbers. These phenomena paralleled a time-dependent rise in the proportion of apoptotic nuclei, as detected by flow cytometry, and of macrophages which had engulfed apoptotic bodies. Apoptotic death of bronchopulmonary cells was observed exclusively following intranasal infection with bacteria reisolated from lungs of infected animals and not with B. pertussis collected after in vitro subculture. Using the terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling technique coupled to fluorescence microscopy and morphological analysis, we established that the apoptotic cells in bronchoalveolar lavage fluids were neutrophils and macrophages. Histological analysis of the lung tissues from B. pertussis-infected mice showed increased numbers of apoptotic cells in the alveolar compartments. Cellular accumulation in bronchoalveolar lavage fluids and apoptosis of alveolar macrophages were significantly attenuated in mice infected with a mutant deficient in the expression of adenylate cyclase-hemolysin, indicating a role of this enzyme in these processes.

Download Full-text