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Adaptability and persistence of the emerging pathogen Bordetella petrii.

Zelazny AM, Ding L, Goldberg JB, Mijares LA, Conlan S, Conville PS, Stock F, Ballentine SJ, Olivier KN, Sampaio EP, Murray PR, Holland SM - PLoS ONE (2013)

Bottom Line: Strains were compared genetically, phenotypically and by antibody recognition from the patient and from inoculated mice.Finally, we characterize one strain that was poorly recognized by the patient's antibodies, due to a defect in the lipopolysaccharide O-antigen, and identify a mutation associated with this phenotype.We propose that B. petrii is remarkably adaptable in vivo, providing a possible connection between immune response and bacterial evasion and supporting infection persistence.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Clinical Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA. azelazny@mail.nih.gov

ABSTRACT
The first described, environmentally isolated, Bordetella petrii was shown to undergo massive genomic rearrangements in vitro. More recently, B. petrii was isolated from clinical samples associated with jaw, ear bone, cystic fibrosis and chronic pulmonary disease. However, the in vivo consequences of B. petrii genome plasticity and its pathogenicity remain obscure. B. petrii was identified from four sequential respiratory samples and a post-mortem spleen sample of a woman presenting with bronchiectasis and cavitary lung disease associated with nontuberculous mycobacterial infection. Strains were compared genetically, phenotypically and by antibody recognition from the patient and from inoculated mice. The successive B. petrii strains exhibited differences in growth, antibiotic susceptibility and recognition by the patient's antibodies. Antibodies from mice inoculated with these strains recapitulated the specificity and strain dependent response that was seen with the patient's serum. Finally, we characterize one strain that was poorly recognized by the patient's antibodies, due to a defect in the lipopolysaccharide O-antigen, and identify a mutation associated with this phenotype. We propose that B. petrii is remarkably adaptable in vivo, providing a possible connection between immune response and bacterial evasion and supporting infection persistence.

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SDS PAGE (A), immunoblots with our patient’s serum (B) and TNF-α induction of B. petrii LPS.(A) Five ml overnight cultures from each bacterial strain (B. petrii 1, B. petrii 3, NCTC 13363 and ATCC BAA-461) were spun 10 min at 8000g to harvest bacterial cells and LPS isolated by using a LPS extraction kit. Extracted LPS samples were separated on a 12% SDS-PAGE and stained using Pro-Q Emerald 300 Lipopolysaccharide gel stain kit. LPS concentrations were measured spectrophotometrically at 205 nm. Control E coli LPS was obtained commercially. I, II and III indicate LPS bands I, II and III respectively, described in the text (B) Immunoblots of our patient’s serum against LPS were performed as described in Fig. 4A. (C) Purified LPS (200 ng/ml) was added to human peripheral blood mononuclear cells (PBMCs) from normal volunteers, and supernatants were collected for cytokine measurements after 20 hours. p<0.001 for NCTC 13363 vs B. petrii 1, B. petrii 3 or BAA-461. Graph shows mean and SEM from three experiments.
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pone-0065102-g005: SDS PAGE (A), immunoblots with our patient’s serum (B) and TNF-α induction of B. petrii LPS.(A) Five ml overnight cultures from each bacterial strain (B. petrii 1, B. petrii 3, NCTC 13363 and ATCC BAA-461) were spun 10 min at 8000g to harvest bacterial cells and LPS isolated by using a LPS extraction kit. Extracted LPS samples were separated on a 12% SDS-PAGE and stained using Pro-Q Emerald 300 Lipopolysaccharide gel stain kit. LPS concentrations were measured spectrophotometrically at 205 nm. Control E coli LPS was obtained commercially. I, II and III indicate LPS bands I, II and III respectively, described in the text (B) Immunoblots of our patient’s serum against LPS were performed as described in Fig. 4A. (C) Purified LPS (200 ng/ml) was added to human peripheral blood mononuclear cells (PBMCs) from normal volunteers, and supernatants were collected for cytokine measurements after 20 hours. p<0.001 for NCTC 13363 vs B. petrii 1, B. petrii 3 or BAA-461. Graph shows mean and SEM from three experiments.

Mentions: Following the differential antibody recognition of B. petrii 3 LPS by the patient and inoculated mice, we compared the SDS-PAGE profiles of LPS from B. petrii 3, B. petrii 1 and reference strains. As shown in Fig. 5A, the LPS from B. petrii 1 and B. petrii 3 show similar profiles for the putative lipid A-inner core (band I) and lipid A-inner core-outer core (band II) components. However, while B. petrii 1 LPS shows a band corresponding to the lipid A-inner core-outer core-O-antigen (band III), the B. petrii 3 LPS lacks this component. The environmentally isolated type strain of B. petrii (ATCC BAA-461) has a band suggestive of O-antigen, while the strain NCTC 13363 does not (Fig. 5A). Interestingly, patient serum recognized putative bands I and II in the LPS of NCTC 13363, but failed to recognize any band of ATCC BAA-461 (Fig. 5B). As expected, the patient recognized O-antigen band of B. petrii 1 LPS as well as band I and II. The later were also detected in B. petrii 3 LPS.


Adaptability and persistence of the emerging pathogen Bordetella petrii.

Zelazny AM, Ding L, Goldberg JB, Mijares LA, Conlan S, Conville PS, Stock F, Ballentine SJ, Olivier KN, Sampaio EP, Murray PR, Holland SM - PLoS ONE (2013)

SDS PAGE (A), immunoblots with our patient’s serum (B) and TNF-α induction of B. petrii LPS.(A) Five ml overnight cultures from each bacterial strain (B. petrii 1, B. petrii 3, NCTC 13363 and ATCC BAA-461) were spun 10 min at 8000g to harvest bacterial cells and LPS isolated by using a LPS extraction kit. Extracted LPS samples were separated on a 12% SDS-PAGE and stained using Pro-Q Emerald 300 Lipopolysaccharide gel stain kit. LPS concentrations were measured spectrophotometrically at 205 nm. Control E coli LPS was obtained commercially. I, II and III indicate LPS bands I, II and III respectively, described in the text (B) Immunoblots of our patient’s serum against LPS were performed as described in Fig. 4A. (C) Purified LPS (200 ng/ml) was added to human peripheral blood mononuclear cells (PBMCs) from normal volunteers, and supernatants were collected for cytokine measurements after 20 hours. p<0.001 for NCTC 13363 vs B. petrii 1, B. petrii 3 or BAA-461. Graph shows mean and SEM from three experiments.
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Related In: Results  -  Collection

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pone-0065102-g005: SDS PAGE (A), immunoblots with our patient’s serum (B) and TNF-α induction of B. petrii LPS.(A) Five ml overnight cultures from each bacterial strain (B. petrii 1, B. petrii 3, NCTC 13363 and ATCC BAA-461) were spun 10 min at 8000g to harvest bacterial cells and LPS isolated by using a LPS extraction kit. Extracted LPS samples were separated on a 12% SDS-PAGE and stained using Pro-Q Emerald 300 Lipopolysaccharide gel stain kit. LPS concentrations were measured spectrophotometrically at 205 nm. Control E coli LPS was obtained commercially. I, II and III indicate LPS bands I, II and III respectively, described in the text (B) Immunoblots of our patient’s serum against LPS were performed as described in Fig. 4A. (C) Purified LPS (200 ng/ml) was added to human peripheral blood mononuclear cells (PBMCs) from normal volunteers, and supernatants were collected for cytokine measurements after 20 hours. p<0.001 for NCTC 13363 vs B. petrii 1, B. petrii 3 or BAA-461. Graph shows mean and SEM from three experiments.
Mentions: Following the differential antibody recognition of B. petrii 3 LPS by the patient and inoculated mice, we compared the SDS-PAGE profiles of LPS from B. petrii 3, B. petrii 1 and reference strains. As shown in Fig. 5A, the LPS from B. petrii 1 and B. petrii 3 show similar profiles for the putative lipid A-inner core (band I) and lipid A-inner core-outer core (band II) components. However, while B. petrii 1 LPS shows a band corresponding to the lipid A-inner core-outer core-O-antigen (band III), the B. petrii 3 LPS lacks this component. The environmentally isolated type strain of B. petrii (ATCC BAA-461) has a band suggestive of O-antigen, while the strain NCTC 13363 does not (Fig. 5A). Interestingly, patient serum recognized putative bands I and II in the LPS of NCTC 13363, but failed to recognize any band of ATCC BAA-461 (Fig. 5B). As expected, the patient recognized O-antigen band of B. petrii 1 LPS as well as band I and II. The later were also detected in B. petrii 3 LPS.

Bottom Line: Strains were compared genetically, phenotypically and by antibody recognition from the patient and from inoculated mice.Finally, we characterize one strain that was poorly recognized by the patient's antibodies, due to a defect in the lipopolysaccharide O-antigen, and identify a mutation associated with this phenotype.We propose that B. petrii is remarkably adaptable in vivo, providing a possible connection between immune response and bacterial evasion and supporting infection persistence.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Clinical Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA. azelazny@mail.nih.gov

ABSTRACT
The first described, environmentally isolated, Bordetella petrii was shown to undergo massive genomic rearrangements in vitro. More recently, B. petrii was isolated from clinical samples associated with jaw, ear bone, cystic fibrosis and chronic pulmonary disease. However, the in vivo consequences of B. petrii genome plasticity and its pathogenicity remain obscure. B. petrii was identified from four sequential respiratory samples and a post-mortem spleen sample of a woman presenting with bronchiectasis and cavitary lung disease associated with nontuberculous mycobacterial infection. Strains were compared genetically, phenotypically and by antibody recognition from the patient and from inoculated mice. The successive B. petrii strains exhibited differences in growth, antibiotic susceptibility and recognition by the patient's antibodies. Antibodies from mice inoculated with these strains recapitulated the specificity and strain dependent response that was seen with the patient's serum. Finally, we characterize one strain that was poorly recognized by the patient's antibodies, due to a defect in the lipopolysaccharide O-antigen, and identify a mutation associated with this phenotype. We propose that B. petrii is remarkably adaptable in vivo, providing a possible connection between immune response and bacterial evasion and supporting infection persistence.

Show MeSH
Related in: MedlinePlus