UPGMA Tree Based on the Analysis of the Pertactin Gene of <i>Bordetella</i> Isolates Used in the MLST Analysis

<p>The DNA segment coding for the extracellular domain of pertactin (P.69) was used for analysis, with the exclusion of the repeat regions 1 and 2. Bootstrap values are shown for the nodes separating the complexes and are based on 500 bootstrap replicates. The scale indicates the genetic distance along the branches. Colors of the branches indicate the four complexes defined by MLST. The number of strains of each branch is shown in boxes, as well as the host distribution.</p

Gene Content of the Differentially Hybridizing Virulence Loci between <i>B. bronchiseptica</i> Complex I and IV, as Determined by CGH

<p>Each column represents one strain. Strain numbers and STs are indicated above the columns. Each row represents one ORF (in <i>B. bronchiseptica</i> RB50 gene order), ORF designations are shown to the right of the rows. In the case of <i>tcfA</i> and <i>prn</i>, the origins of the probes are indicated between parentheses. The BP probe of <i>tcfA</i> was 100% similar to <i>B. pertussis</i> Tohama and 85.1% similar to <i>B. bronchiseptica</i> RB50. The BP <i>prn</i> probe was 100% similar to <i>B. pertussis</i> Tohama and 86% similar to <i>B. parapertussis</i> 12822 and <i>B. bronchiseptica</i> RB50. The BB/BPP <i>prn</i> probes were both 100% similar to <i>B. parapertussis</i> 12822 (BPP) and <i>B. bronchiseptica</i> RB50 (BB) and 86% similar to <i>B. pertussis</i> Tohama. The yellow-black-blue color scale indicates the hybridization value relative to the reference; references are <i>B. bronchiseptica</i> RB50, <i>B. parapertussis</i> 12822, and <i>B. pertussis</i> Tohama. For <i>B. bronchiseptica</i> RB50 and <i>B. pertussis</i> Tohama, the data in the figure are based on the genomic sequences. Yellow indicates decreased hybridization, black indicates hybridization values comparative to the references, and blue indicates gene duplications. Intermediate values indicate partial deletions or sequence divergence. Missing data are represented in gray.</p

Model of the Evolution of the Mammalian Bordetellae

<p>The bar on the left indicates increasing degrees of adaptation to the human host. Arrows indicate descent; double arrows between complexes indicate possible within-host immune competition. In boxes, genetic events are shown that may have played a role in speciation and niche adaptation. Numbers between parentheses refer to references. See text for details.</p

Expression of LPS by <i>B. bronchiseptica</i> Complex I and Complex IV Strains and Gene Content Variation at the LPS Biosynthesis Locus

<div><p>(A) Top: Electrophoretic LPS profiles obtained by tricine-SDS-PAGE and silver staining. Middle: Western blot of the same samples with mAb 36G3, which detects band A. Bottom: Western blot of the same samples with mAb BL8, which detects band B.</p><p>(B) Gene content of the LPS biosynthesis locus as determined by CGH. See <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.0010045#ppat-0010045-g003" target="_blank">Figure 3</a> for details. For <i>B. bronchiseptica</i> RB50 and <i>B. pertussis</i> Tohama, the data in the figure are based on the genomic sequences. The genes <i>wbmPQRSTU</i> represent an alternative LPS O-antigen biosynthesis sublocus that is orthologous to the genes found in <i>B. parapertussis</i> 12822 [<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.0010045#ppat-0010045-b010" target="_blank">10</a>] and <i>B. bronchiseptica</i> C7635E [<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.0010045#ppat-0010045-b026" target="_blank">26</a>]. LPS genetic profiles as described in the text are indicated at the top of the columns. Color scale as in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.0010045#ppat-0010045-g003" target="_blank">Figure 3</a>. Missing data are represented in gray.</p></div

Minimum Spanning Tree of <i>B. bronchiseptica, B. pertussis,</i> and <i>B. parapertussis</i>

<p>The tree was based on the sequence of seven housekeeping genes. Individual genes were split into five subloci, and a categorical clustering was performed. In the minimum spanning tree, sequence types sharing the highest number of single locus variants were connected first. Each circle represents a sequence type (ST) the size of which is related to the number of isolates within that particular ST. Colors within circles indicate host distribution. The numbers between connected STs represent the number of different subloci between those STs. The clonal complexes (I, II, III, and IV) are indicated by colored strips between connected STs. ST16 (<i>B. bronchiseptica</i> complex I) harbors the <i>B. parapertussis_ov</i> strains. STs containing strains of which the genome has been sequenced (<i>B. pertussis</i> Tohama, <i>B. parapertussis</i> 12822 or <i>B. bronchiseptica</i> RB50) are indicated by a thickset, dashed line. The distribution of the insertion sequence elements IS<i>481</i>, IS<i>1001</i>, IS<i>1002,</i> and IS<i>1663</i> is shown in boxes (see also <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.0010045#ppat-0010045-st001" target="_blank">Table S1</a>); numbers between parentheses indicate the percentage of strains that contained the ISE as determined by PCR amplification. The divergence times between <i>B. bronchiseptica</i> complexes I and IV and <i>B. pertussis</i> complex II are shown.</p

The role of Toll-like receptor-4 in pertussis vaccine-induced immunity-0

Injected with 1/5 human dose (HD) wP, aP, or adjuvant (C), twice before intranasal infection. Three and seven days after challenge lungs were excised, and the number of viable was determined in right lung lobes. Each symbol represents the number of bacteria in the lung of an individual mouse; horizontal lines represent the group average. Non-boxed -values: compared to the indicated treatment group (same strain and day after infection). Boxed -values: compared to the wild-type strain (same treatment and day after infection). ANOVA followed by Bonferroni post-hoc test. A single representative experiment of 2 is shown.<p><b>Copyright information:</b></p><p>Taken from "The role of Toll-like receptor-4 in pertussis vaccine-induced immunity"</p><p>http://www.biomedcentral.com/1471-2172/9/21</p><p>BMC Immunology 2008;9():21-21.</p><p>Published online 22 May 2008</p><p>PMCID:PMC2409298.</p><p></p

The role of Toll-like receptor-4 in pertussis vaccine-induced immunity-1

Uvant (C), twice before intranasal infection. Two hours before challenge, and three and seven days after challenge blood was taken. Test and positive control sera were tested for Ptx-, FHA-, and Prn-specific IgG. Each symbol represents the serum level of an individual mouse; horizontal lines represent the group average. (#) increased compared to the adjuvant control (same strain and time point; < 0.001), (+) increased compared to wP-vaccinated mice (same strain and time point; < 0.001). Non-boxed -values: compared to the indicated group (same strain and time point), boxed -values: compared to the wild-type strain (same treatment and time point), and stippled-boxed P-values: compared to the pre-challenge level (same strain and treatment). ANOVA followed by Bonferroni post-hoc test. A single representative experiment of 2 is shown.<p><b>Copyright information:</b></p><p>Taken from "The role of Toll-like receptor-4 in pertussis vaccine-induced immunity"</p><p>http://www.biomedcentral.com/1471-2172/9/21</p><p>BMC Immunology 2008;9():21-21.</p><p>Published online 22 May 2008</p><p>PMCID:PMC2409298.</p><p></p

The role of Toll-like receptor-4 in pertussis vaccine-induced immunity-5

Injected with 1/5 human dose (HD) wP, aP, or adjuvant (C), twice before intranasal infection. Three and seven days after challenge lungs were excised, and the number of viable was determined in right lung lobes. Each symbol represents the number of bacteria in the lung of an individual mouse; horizontal lines represent the group average. Non-boxed -values: compared to the indicated treatment group (same strain and day after infection). Boxed -values: compared to the wild-type strain (same treatment and day after infection). ANOVA followed by Bonferroni post-hoc test. A single representative experiment of 2 is shown.<p><b>Copyright information:</b></p><p>Taken from "The role of Toll-like receptor-4 in pertussis vaccine-induced immunity"</p><p>http://www.biomedcentral.com/1471-2172/9/21</p><p>BMC Immunology 2008;9():21-21.</p><p>Published online 22 May 2008</p><p>PMCID:PMC2409298.</p><p></p

The role of Toll-like receptor-4 in pertussis vaccine-induced immunity-2

Uvant (C), twice before intranasal infection. Two hours before challenge, and three and seven days after challenge mice were weighed. Data are indicated as mean ± SEM (N = 6). Non-boxed -values: compared to the adjuvant control (same strain and day after challenge). Boxed -value: compared to the wild-type strain (same treatment and day after challenge). ANOVA followed by Bonferroni post-hoc test. A single representative experiment of 2 is shown.<p><b>Copyright information:</b></p><p>Taken from "The role of Toll-like receptor-4 in pertussis vaccine-induced immunity"</p><p>http://www.biomedcentral.com/1471-2172/9/21</p><p>BMC Immunology 2008;9():21-21.</p><p>Published online 22 May 2008</p><p>PMCID:PMC2409298.</p><p></p