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Virulence Dec 2021and are major pathogens in pigs, which are frequently isolated from co-infections in the respiratory tract and contribute to the porcine respiratory disease complex...
and are major pathogens in pigs, which are frequently isolated from co-infections in the respiratory tract and contribute to the porcine respiratory disease complex (PRDC). Despite the high impact of co-infections on respiratory diseases of swine (and other hosts), very little is known about pathogen-pathogen-host interactions and the mechanisms of pathogenesis. In the present study, we established a porcine precision-cut lung slice (PCLS) model to analyze the effects of infection on adherence, colonization, and cytotoxic effects of . We hypothesized that induction of ciliostasis by a clinical isolate of may promote subsequent infection with a virulent serotype 2 strain. To investigate this theory, we monitored the ciliary activity by light microscopy, measured the release of lactate dehydrogenase, and calculated the number of PCLS-associated bacteria. To study the role of the pore-forming toxin suilysin (SLY) in -induced cytotoxicity, we included a SLY-negative isogenic mutant and the complemented mutant strain. Furthermore, we analyzed infected PCLS by histopathology, immunofluorescence microscopy, and field emission scanning electron microscopy. Our results showed that pre-infection with promoted adherence, colonization, and, as a consequence of the increased colonization, the cytotoxic effects of , probably by reduction of the ciliary activity. Moreover, cytotoxicity induced by is strictly dependent on the presence of SLY. Though the underlying molecular mechanisms remain to be fully clarified, our results clearly support the hypothesis that paves the way for infection.
Topics: Animals; Bacterial Adhesion; Bordetella bronchiseptica; Cilia; Coinfection; Hemolysin Proteins; Host-Pathogen Interactions; In Vitro Techniques; Lung; Streptococcus suis; Swine; Swine Diseases
PubMed: 33372837
DOI: 10.1080/21505594.2020.1858604 -
Toxins Sep 2017and are the causal agents of whooping cough in humans. They produce diverse virulence factors, including adenylate cyclase-hemolysin (AC-Hly), a secreted toxin of the...
Characterization of Post-Translational Modifications and Cytotoxic Properties of the Adenylate-Cyclase Hemolysin Produced by Various Bordetella pertussis and Bordetella parapertussis Isolates.
and are the causal agents of whooping cough in humans. They produce diverse virulence factors, including adenylate cyclase-hemolysin (AC-Hly), a secreted toxin of the repeat in toxins (RTX) family with cyclase, pore-forming, and hemolytic activities. Post-translational modifications (PTMs) are essential for the biological activities of the toxin produced by . In this study, we compared AC-Hly toxins from various clinical isolates of and , focusing on (i) the genomic sequences of genes, (ii) the PTMs of partially purified AC-Hly, and (iii) the cytotoxic activity of the various AC-Hly toxins. The genes encoding the AC-Hly toxins of and displayed very limited polymorphism in each species. Most of the sequence differences between the two species were found in the C-terminal part of the protein. Both toxins harbored PTMs, mostly corresponding to palmitoylations of the lysine 860 residue and palmoylations and myristoylations of lysine 983 for and AC-Hly and palmitoylations of lysine 894 and myristoylations of lysine 1017 for AC-Hly. Purified AC-Hly from was cytotoxic to macrophages, whereas that from was not.
Topics: Adenylate Cyclase Toxin; Bordetella parapertussis; Bordetella pertussis; Cell Line; Macrophages; Protein Processing, Post-Translational
PubMed: 28954396
DOI: 10.3390/toxins9100304 -
Microbiology Spectrum Oct 2021Noncoding small RNAs (sRNAs) are crucial for the posttranscriptional regulation of gene expression in all organisms and are known to be involved in the regulation of...
Noncoding small RNAs (sRNAs) are crucial for the posttranscriptional regulation of gene expression in all organisms and are known to be involved in the regulation of bacterial virulence. In the human pathogen Bordetella pertussis, which causes whooping cough, virulence is controlled primarily by the master two-component system BvgA (response regulator)/BvgS (sensor kinase). In this system, BvgA is phosphorylated (Bvg mode) or nonphosphorylated (Bvg mode), with global transcriptional differences between the two. B. pertussis also carries the bacterial sRNA chaperone Hfq, which has previously been shown to be required for virulence. Here, we conducted transcriptomic analyses to identify possible B. pertussis sRNAs and to determine their BvgAS dependence using transcriptome sequencing (RNA-seq) and the prokaryotic sRNA prediction program ANNOgesic. We identified 143 possible candidates (25 Bvg mode specific and 53 Bvg mode specific), of which 90 were previously unreported. Northern blot analyses confirmed all of the 10 ANNOgesic candidates that we tested. Homology searches demonstrated that 9 of the confirmed sRNAs are highly conserved among B. pertussis, Bordetella parapertussis, and Bordetella bronchiseptica, with one that also has homologues in other species of the family. Using coimmunoprecipitation with a B. pertussis FLAG-tagged Hfq, we demonstrated that 3 of the sRNAs interact directly with Hfq, which is the first identification of sRNA binding to B. pertussis Hfq. Our study demonstrates that ANNOgesic is a highly useful tool for the identification of sRNAs in this system and that its combination with molecular techniques is a successful way to identify various BvgAS-dependent and Hfq-binding sRNAs. Noncoding small RNAs (sRNAs) are crucial for posttranscriptional regulation of gene expression in all organisms and are known to be involved in the regulation of bacterial virulence. We have investigated the presence of sRNAs in the obligate human pathogen B. pertussis, using transcriptome sequencing (RNA-seq) and the recently developed prokaryotic sRNA search program ANNOgesic. This analysis has identified 143 sRNA candidates (90 previously unreported). We have classified their dependence on the B. pertussis two-component system required for virulence, namely, BvgAS, based on their expression in the presence/absence of the phosphorylated response regulator BvgA, confirmed several by Northern analyses, and demonstrated that 3 bind directly to B. pertussis Hfq, the RNA chaperone involved in mediating sRNA effects. Our study demonstrates the utility of combining RNA-seq, ANNOgesic, and molecular techniques to identify various BvgAS-dependent and Hfq-binding sRNAs, which may unveil the roles of sRNAs in pertussis pathogenesis.
Topics: Bacterial Proteins; Bordetella bronchiseptica; Bordetella parapertussis; Bordetella pertussis; Computational Biology; Gene Expression Profiling; Gene Expression Regulation, Bacterial; Host Factor 1 Protein; RNA, Small Untranslated; Software; Transcription Factors; Transcriptome; Virulence; Virulence Factors, Bordetella
PubMed: 34550019
DOI: 10.1128/Spectrum.00044-21 -
Journal of Clinical Microbiology Mar 1998PCR, using primers Plp1 and Plp2, was evaluated for the detection of DNA from Bordetella pertussis in bacterial strains and in nasopharyngeal samples from patients with... (Comparative Study)
Comparative Study
PCR, using primers Plp1 and Plp2, was evaluated for the detection of DNA from Bordetella pertussis in bacterial strains and in nasopharyngeal samples from patients with a cough lasting at least 7 days. The assay could detect DNA from 6 CFU of B. pertussis/10 microl of sample. Results of the PCR assay were compared with those of cultures, a determination of serum antibodies against pertussis toxin and filamentous hemagglutinin, and a clinical evaluation of 2,442 coughing episodes. The overall sensitivity of PCR was 65% (623 of 956), which was higher than the sensitivity of cultures (58%) (P < 0.001). Factors influencing the sensitivity of PCR were the interval between the onset of symptoms and sampling and the vaccination status of the patient. The specificity of PCR was 98% (1,451 of 1,486). The positive and negative predictive values were 95 and 81%, respectively. Parapertussis PCR, using primers BPPA and BPPZ, was positive in 11 of 18 culture-positive cases and was confirmed by serology in another 4 cases. In conclusion, PCR is a valuable complement to cultures and can probably replace cultures for diagnosis of B. pertussis and Bordetella parapertussis infections.
Topics: Antibodies, Bacterial; Bordetella; Bordetella Infections; Bordetella pertussis; DNA Primers; DNA, Bacterial; Diphtheria-Tetanus-Pertussis Vaccine; Enzyme-Linked Immunosorbent Assay; Evaluation Studies as Topic; Humans; Infant; Nasopharynx; Polymerase Chain Reaction; Predictive Value of Tests; Sensitivity and Specificity; Toxoids; Whooping Cough
PubMed: 9508295
DOI: 10.1128/JCM.36.3.679-683.1998 -
Annals of Laboratory Medicine Jul 2021
Review
Topics: Asia; Bordetella; Bordetella Infections; Humans; Pneumonia
PubMed: 33536366
DOI: 10.3343/alm.2021.41.4.439 -
Epidemiology and Infection Feb 2000Fifty-seven bacterial isolates previously identified as Bordetella avium or B. hinzii were characterized by restriction enzyme analysis (REA) and/or ribotyping. Twenty...
Fifty-seven bacterial isolates previously identified as Bordetella avium or B. hinzii were characterized by restriction enzyme analysis (REA) and/or ribotyping. Twenty restriction endonucleases were evaluated for REA. Digestion of chromosomal DNA from the 42 B. avium and 15 B. hinzii isolates with HinfI produced 8 and 7 distinct fingerprint profiles, respectively. Digestion with DdeI further discriminated these Bordetella species and produced 12 fingerprint profiles for B. avium and 4 profiles of B. hinzii. In addition, B. avium isolates were clearly distinguishable from B. hinzii isolates by ribotyping with the restriction endonuclease PvuII. The ribotype patterns of these two species of Bordetella were unique when compared to previously reported ribotype patterns for B. bronchiseptica isolates. Since it was possible to discern differences among isolates within each Bordetella species by REA analysis, we suggest that REA could be used in developing a typing system based on the fingerprint profiles generated.
Topics: Bacterial Typing Techniques; Bordetella; DNA Fingerprinting; DNA Restriction Enzymes; DNA, Bacterial; DNA, Ribosomal; Humans; Polymorphism, Restriction Fragment Length; Prohibitins; Restriction Mapping
PubMed: 10722134
DOI: 10.1017/s0950268899003337 -
Infection and Immunity Dec 2004Surfactant proteins A (SP-A) and D (SP-D) play an important role in the innate immune defenses of the respiratory tract. SP-A binds to the lipid A region of...
Interactions of pulmonary collectins with Bordetella bronchiseptica and Bordetella pertussis lipopolysaccharide elucidate the structural basis of their antimicrobial activities.
Surfactant proteins A (SP-A) and D (SP-D) play an important role in the innate immune defenses of the respiratory tract. SP-A binds to the lipid A region of lipopolysaccharide (LPS), and SP-D binds to the core oligosaccharide region. Both proteins induce aggregation, act as opsonins for neutrophils and macrophages, and have direct antimicrobial activity. Bordetella pertussis LPS has a branched core structure and a nonrepeating terminal trisaccharide. Bordetella bronchiseptica LPS has the same structure, but lipid A is palmitoylated and there is a repeating O-antigen polysaccharide. The ability of SP-A and SP-D to agglutinate and permeabilize wild-type and LPS mutants of B. pertussis and B. bronchiseptica was examined. Previously, wild-type B. pertussis was shown to resist the effects of SP-A; however, LPS mutants lacking the terminal trisaccharide were susceptible to SP-A. In this study, SP-A was found to aggregate and permeabilize a B. bronchiseptica mutant lacking the terminal trisaccharide, while wild-type B. bronchiseptica and mutants lacking only the palmitoyl transferase or O antigen were resistant to SP-A. Wild-type B. pertussis and B. bronchiseptica were both resistant to SP-D; however, LPS mutants of either strain lacking the terminal trisaccharide were aggregated and permeabilized by SP-D. We conclude that the terminal trisaccharide protects Bordetella species from the bactericidal functions of SP-A and SP-D. The O antigen and palmitoylated lipid A of B. bronchiseptica play no role in this resistance.
Topics: Animals; Anti-Bacterial Agents; Bordetella bronchiseptica; Bordetella pertussis; Cell Membrane Permeability; Lipopolysaccharides; Macrophages, Alveolar; Phagocytosis; Pulmonary Surfactant-Associated Protein A; Pulmonary Surfactant-Associated Protein D; Rats
PubMed: 15557636
DOI: 10.1128/IAI.72.12.7124-7130.2004 -
Clinical and Vaccine Immunology : CVI Nov 2016Pertussis is a severe respiratory disease caused by infection with the bacterial pathogen Bordetella pertussis The disease affects individuals of all ages but is...
Pertussis is a severe respiratory disease caused by infection with the bacterial pathogen Bordetella pertussis The disease affects individuals of all ages but is particularly severe and sometimes fatal in unvaccinated young infants. Other Bordetella species cause diseases in humans, animals, and birds. Scientific, clinical, public health, vaccine company, and regulatory agency experts on these pathogens and diseases gathered in Buenos Aires, Argentina from 5 to 8 April 2016 for the 11th International Bordetella Symposium to discuss recent advances in our understanding of the biology of these organisms, the diseases they cause, and the development of new vaccines and other strategies to prevent these diseases. Highlights of the meeting included pertussis epidemiology in developing nations, genomic analysis of Bordetella biology and evolution, regulation of virulence factor expression, new model systems to study Bordetella biology and disease, effects of different vaccines on immune responses, maternal immunization as a strategy to prevent newborn disease, and novel vaccine development for pertussis. In addition, the group approved the formation of an International Bordetella Society to promote research and information exchange on bordetellae and to organize future meetings. A new Bordetella.org website will also be developed to facilitate these goals.
Topics: Animals; Argentina; Bacterial Outer Membrane Proteins; Bordetella pertussis; Humans; Infant; Pertussis Vaccine; Vaccination; Virulence Factors, Bordetella; Whooping Cough
PubMed: 27655886
DOI: 10.1128/CVI.00388-16 -
Microbes and Infection Jan 2001The genus Bordetella comprises seven species with pathogenic potential for different host organisms. This article attempts to review our current knowledge about the... (Review)
Review
The genus Bordetella comprises seven species with pathogenic potential for different host organisms. This article attempts to review our current knowledge about the systematics and evolution of this important group of pathogens, their relationship to environmental microorganisms and about molecular mechanisms of host adaptation.
Topics: Animals; Biological Evolution; Bordetella; Genome, Bacterial; Humans; Phylogeny; Species Specificity
PubMed: 11226855
DOI: 10.1016/s1286-4579(00)01353-8 -
Clinical Microbiology and Infection :... Sep 2012Bordetella pertussis and Bordetella parapertussis are closely related bacterial agents of whooping cough. Whole-cell pertussis (wP) vaccine was introduced in France in...
Bordetella pertussis and Bordetella parapertussis are closely related bacterial agents of whooping cough. Whole-cell pertussis (wP) vaccine was introduced in France in 1959. Acellular pertussis (aP) vaccine was introduced in 1998 as an adolescent booster and was rapidly generalized to the whole population, changing herd immunity by specifically targeting the virulence of the bacteria. We performed a temporal analysis of all French B. pertussis and B. parapertussis isolates collected since 2000 under aP vaccine pressure, using pulsed-field gel electrophoresis (PFGE), genotyping and detection of expression of virulence factors. Particular isolates were selected according to their different phenotype and PFGE type and their characteristics were analysed using the murine model of respiratory infection and in vitro cell cytotoxic assay. Since the introduction of the aP vaccines there has been a steady increase in the number of B. pertussis and B. parapertussis isolates collected that are lacking expression of pertactin. These isolates seem to be as virulent as those expressing all virulence factors according to animal and cellular models of infection. Whereas wP vaccine-induced immunity led to a monomorphic population of B. pertussis, aP vaccine-induced immunity enabled the number of circulating B. pertussis and B. parapertussis isolates not expressing virulence factors to increase, sustaining our previous hypothesis.
Topics: Animals; Bacterial Outer Membrane Proteins; Blotting, Western; Bordetella Infections; Bordetella parapertussis; Bordetella pertussis; Disease Models, Animal; Electrophoresis, Gel, Pulsed-Field; Evolution, Molecular; Genotype; Humans; Macrophages; Mice; Pertussis Vaccine; Virulence Factors, Bordetella; Whooping Cough
PubMed: 22717007
DOI: 10.1111/j.1469-0691.2012.03925.x