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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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Related in: MedlinePlus

Immunoblots with patient’s serum against her own and reference strains.An aliquot containing 10 µg of protein from the soluble (A) and insoluble (B) fractions obtained from B. petrii strains was electrophoresed on 12% SDS–PAGE, and then transferred to PVDF membrane. The membrane was incubated with the patient serum (1∶5000 dilution) obtained ∼2 months after isolation of B. petrii 3 and then horseradish peroxidase conjugated sheep anti-human IgG (1∶10000). The blots were developed using the enhanced chemiluminescence kit. B. petrii 4, B. petrii 4b, B. petrii 4c and B. petrii 4d refer to four different colonies obtained from the primary isolation plate of B. petrii 4. Fig. 2B had a shorter exposure time than Fig. 2A, so bands could be better visualized. If using similar exposure times, the intensity of the bands in Fig. 2B is 48% higher than in Fig. 2A (as determined by densitometric analysis).
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pone-0065102-g002: Immunoblots with patient’s serum against her own and reference strains.An aliquot containing 10 µg of protein from the soluble (A) and insoluble (B) fractions obtained from B. petrii strains was electrophoresed on 12% SDS–PAGE, and then transferred to PVDF membrane. The membrane was incubated with the patient serum (1∶5000 dilution) obtained ∼2 months after isolation of B. petrii 3 and then horseradish peroxidase conjugated sheep anti-human IgG (1∶10000). The blots were developed using the enhanced chemiluminescence kit. B. petrii 4, B. petrii 4b, B. petrii 4c and B. petrii 4d refer to four different colonies obtained from the primary isolation plate of B. petrii 4. Fig. 2B had a shorter exposure time than Fig. 2A, so bands could be better visualized. If using similar exposure times, the intensity of the bands in Fig. 2B is 48% higher than in Fig. 2A (as determined by densitometric analysis).

Mentions: To determine whether the patient generated antibodies against components of B. petrii, immunoblots were performed using serum from the patient. A strong response was seen against soluble fractions of bacterial extracts from B. petrii 1, B. petrii 2, B. petrii 4 and B. petrii 5 but not those from B. petrii 3 or the reference strains (Fig. 2A). A dominant band with apparent MW ∼14–28 KDa, a high molecular weight band (>98 KDa) and two smaller bands (∼6 KDa) were seen in B. petrii 1, B. petrii 2, and B. petrii 5 profiles. B. petrii 3 profile included the two smaller bands but not the dominant 14–28 KDa nor the >98 KDa bands, while only the later was missing in B. petrii 4. Screening additional colonies of B. petrii 4 yielded comparable results.


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)

Immunoblots with patient’s serum against her own and reference strains.An aliquot containing 10 µg of protein from the soluble (A) and insoluble (B) fractions obtained from B. petrii strains was electrophoresed on 12% SDS–PAGE, and then transferred to PVDF membrane. The membrane was incubated with the patient serum (1∶5000 dilution) obtained ∼2 months after isolation of B. petrii 3 and then horseradish peroxidase conjugated sheep anti-human IgG (1∶10000). The blots were developed using the enhanced chemiluminescence kit. B. petrii 4, B. petrii 4b, B. petrii 4c and B. petrii 4d refer to four different colonies obtained from the primary isolation plate of B. petrii 4. Fig. 2B had a shorter exposure time than Fig. 2A, so bands could be better visualized. If using similar exposure times, the intensity of the bands in Fig. 2B is 48% higher than in Fig. 2A (as determined by densitometric analysis).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3672207&req=5

pone-0065102-g002: Immunoblots with patient’s serum against her own and reference strains.An aliquot containing 10 µg of protein from the soluble (A) and insoluble (B) fractions obtained from B. petrii strains was electrophoresed on 12% SDS–PAGE, and then transferred to PVDF membrane. The membrane was incubated with the patient serum (1∶5000 dilution) obtained ∼2 months after isolation of B. petrii 3 and then horseradish peroxidase conjugated sheep anti-human IgG (1∶10000). The blots were developed using the enhanced chemiluminescence kit. B. petrii 4, B. petrii 4b, B. petrii 4c and B. petrii 4d refer to four different colonies obtained from the primary isolation plate of B. petrii 4. Fig. 2B had a shorter exposure time than Fig. 2A, so bands could be better visualized. If using similar exposure times, the intensity of the bands in Fig. 2B is 48% higher than in Fig. 2A (as determined by densitometric analysis).
Mentions: To determine whether the patient generated antibodies against components of B. petrii, immunoblots were performed using serum from the patient. A strong response was seen against soluble fractions of bacterial extracts from B. petrii 1, B. petrii 2, B. petrii 4 and B. petrii 5 but not those from B. petrii 3 or the reference strains (Fig. 2A). A dominant band with apparent MW ∼14–28 KDa, a high molecular weight band (>98 KDa) and two smaller bands (∼6 KDa) were seen in B. petrii 1, B. petrii 2, and B. petrii 5 profiles. B. petrii 3 profile included the two smaller bands but not the dominant 14–28 KDa nor the >98 KDa bands, while only the later was missing in B. petrii 4. Screening additional colonies of B. petrii 4 yielded comparable results.

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