12 research outputs found

    Functional networks most significanty modulated in PPD-B stimulated PBMC from vaccinated/protected calves.

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    <p>Visualisation of the trend and significance of each network: Red bars = up-regulation; blue bars = down-regulation of network. Horizontal bar: p = 0.05.</p

    Results of RNA-Seq analysis.

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    <p>A. Signficantly modulated genes in the three treatment groups. B. Venn diagrams of genes significantly up-regulated and (C) down-regulated genes after vaccination but prior to <i>M. bovis</i> challenge. A. Fold change compared to unstimulated PBMC (medium controls) of PPD-B stimulated PBMC compared to medium controls from unvaccinated, naïve calves (group 1), vaccinated/non-protected (group 2), and vaccinated/protected calves (group 3).</p

    Gene expression in PPDB-stimulated PBMC from BCG vaccinated and control cattle.

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    <p>A. Protective efficacy after <i>M. bovis</i> challenge determined by pathology scoring. <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1003077#s2" target="_blank">Results</a> are expressed as total pathology scores <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1003077#ppat.1003077-Vordermeier1" target="_blank">[5]</a>. Filled circles, unvaccinated control calves; open triangles, BCG vaccinated calves. (B, C). Transcription of the genes expressing IFN-γ (B) and IL-22 (C) following <i>in vitro</i> stimulation. PBMC were collected from BCG vaccinated (filled symbols) and controls (open symbols) before challenge (week -1) and after challenge with <i>M. bovis</i> at weeks 2, 4 and 8, and stimulated with PPD-B for 24 hours. cDNA was prepared and gene expression determined by RT-qPCR. Data are expressed as log10 relative expression levels compared to non-stimulated cells. Statistical analysis: 2-way ANOVA with Bonferroni post test, * P<0.05.</p

    BCG-vaccinated and control cattle samples mapped to <i>ifn-γ</i> and <i>il-22</i> genes.

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    <p>Visualization by IGB of RNA sequencing reads of representative PPD-B stimulated PBMC from vaccinated-protected, vaccinate-un-protected and non-vaccinated control cattle. Y-axis shows the number of reads covering each base along the transcript in RPKM expression values for each sample. Black track: unvaccinated control cattle; red track: vaccinated/un-protected cattle and blue track: vaccinated/protected cattle. The schematic representation of transcript for (A) <i>ifn-γ</i> and (B) <i>il-22</i> is show in green at the bottom of the each figure; the boxes show the exons of the gene.</p

    Protection and <i>in vitro</i> IFN-γ responses prior to challenge.

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    <p>A. Individual pathology scores are shown for the animals used in this study. Naïve animals = no vaccination, Vacc/NP = vaccinated calves that were not protected; Vacc/P = vaccinated calves that were protected. B. Correlation of IFN-γ protein production in culture supernatants measured by Bovigam ELISA (y-axis) and <i>ifn-γ</i> gene expression as determined by deep sequencing (x-axis). Data are shown from PPD-B stimulated PBMC from individual animals. Supernatants and RNA were prepared after 24 h culture.</p

    Schematic representation of the genes involved in the cytokine-cytokine receptor interaction.

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    <p>Colour codes indicate genes that were significantly modulated in vaccinated/protected calves prior to <i>M. bovis</i> challenge. Red colour: up-regulated genes; blue colour: down-regulated genes. Darkness of colour indicates level of gene modification.</p

    Spleen gene signature after 3 and 14 days after infection with <i>M. bovis</i>.

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    <p>The global transcriptional response in spleen cells of mice infected with <i>M. bovis</i> was compared to responses in uninfected mice. Genes were considered significantly modulated when their corrected p-values were below 0.05 with more than 2-fold change of expression at both time points. After 3 and 14 days post-infection (blue square and red squares, respectively at the bottom of graph), the mice were euthanized and their splenocytes were stimulated in vitro for 3 days with M7 protein pool (<i>see </i><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030626#s4" target="_blank"><i>Materials and Methods</i></a>). Black squares: Naïve control mice. Unsupervised hierarchical cluster was performed using a centroid linkage with a Person centered measure showing that 618 of these genes were modulated at both time points (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030626#pone.0030626.s001" target="_blank">table S1</a> for list of these genes, with genes significantly modulated at both time points highlighted in bold).</p

    Pulmonary gene signature after 14 days after infection with <i>M. bovis</i>.

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    <p>The global transcriptional response in the lung of mice infected with <i>M. bovis</i> was compared the response of uninfected mice. Genes were considered significant when their correct p-value were below 0.05 with more than 2-fold change of expression. After 14 days post-infection the mice were euthanized and their lung cells were stimulated in vitro for 3 days with M7 protein pool (<i>see </i><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030626#s4" target="_blank"><i>Materials and Methods</i></a>). Unsupervised hierarchical cluster was performed using a centroid linkage with a Person centered measure showing that 282 of these genes were modulated (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030626#pone.0030626.s002" target="_blank">table S2</a> for list of these genes).</p

    Expression of the most up-regulated genes found in the murine model in cattle using PBMC from uninfected (bTb-free, n = 9) and naturally with <i>M. bovis</i> infected cows (bTb, n = 11).

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    <p>The data are represented as mean (± SEM) fold changes in expression after stimulation of PBMC with bovine PPD-B. Significance level for comparison of results in bTb-free and infected animals: p -value≤0.0033 (*).</p>a<p>Used as positive control.</p><p>NA, not applicable.</p

    Functional networks (A) and canonical pathways (B) most significantly modulated in lung cells 14 days after <i>M. bovis</i> infection.

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    <p>Visualization of the trend and significance in the regulation of each network and pathway. Dark blue: all genes represented in a network. Light blue: genes that were up-regulated in a network. Cyan: gene those were down-regulated in a network. Fisher's exact test threshold value of p≤0.05.</p
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