17 research outputs found

    sCD14 exerts bimodal effects in acute lung inflammation depending on the dose of S-LPS.

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    <p>WT and CD14KO mice were treated intranasally with 10 Β΅g S-LPS (left panel) or 0.1 Β΅g S-LPS (right panel) and 10 Β΅g sCD14 was administered simultaneously with S-LPS to groups of CD14KO mice. Six hours after LPS (and sCD14) administration, BALF was isolated and analyzed for PMN counts (A, B), TNF levels (C, D) and LIX levels (ER, F). Eight to nine mice were used per group. Data are are mean Β± SEM. *, P<0.05; **, P<0.01; ***, P<0001 versus WT mice; ##, P<0.01; ###, P<0.001 versus CD14KO mice.</p

    Pulmonary CD14 partially diminishes lung inflammation by high dose R-LPS, but enhances lung inflammation by low dose R-LPS.

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    <p>Mice (nβ€Š=β€Š6–9) were treated intranasally with 10 Β΅g R-LPS (left panel), 1 Β΅g R-LPS (middle panel) or 0.1 Β΅g R-LPS (right panel). Six hours after LPS administration, BALF was isolated and analysed for PMN counts (A–C), TNF levels (D–F) and LIX levels (G–I). Data are mean Β± SEM. *, P<0.05; **, P<0.01; ***, P<0001 versus WT mice.</p

    Pulmonary CD14 diminishes lung inflammation by high dose S-LPS, but enhances lung inflammation by low dose S-LPS.

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    <p>Mice (nβ€Š=β€Š7–9) were treated intranasally with 10 Β΅g S-LPS (left panel), 1 Β΅g S-LPS (middle panel) or 0.1 Β΅g S-LPS (right panel). Six hours later BALF was isolated and analysed for PMN counts (A–C), TNF levels (D–F) and LIX levels (G–I). Data are mean Β± SEM. **, P<0.01; ***, P<0001 versus WT mice.</p

    S-LPS induces sCD14 release in the lung in a dose dependent manner.

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    <p>sCD14 was measured in BALF obtained from WT mice 6 hours after intranasal administration of different doses (10–0.1 Β΅g) of S-LPS. Eight to nine mice were used per group. Data are mean Β± SEM. Dotted line represents the mean value of sCD14 in BALF of naive mice.</p

    Inflammatory response in <i>LysM-MyD88<sup>βˆ’/βˆ’</sup></i> and <i>Tie2-MyD88<sup>βˆ’/βˆ’</sup></i> during <i>K. pneumonia</i> pulmonary tract infection.

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    <p>Control, LysM-MyD88<sup>βˆ’/βˆ’</sup> and Tie2-MyD88<sup>βˆ’/βˆ’</sup> mice were inoculated with ∼6Γ—10<sup>3</sup> CFU <i>K. pneumoniae</i> and sacrificed 24 hours later. Homogenates were prepared from right lungs. Cytokine and chemokine levels are presented in pg/ml lung homogenate or plasma. Data are mean (SE) of 5–8 mice per group.</p><p>*<i> p</i><0.05,</p><p>** <i>p</i><0.01 vs control mice.</p><p>Bdβ€Š=β€Šbelow detection.</p><p>Inflammatory response in <i>LysM-MyD88<sup>βˆ’/βˆ’</sup></i> and <i>Tie2-MyD88<sup>βˆ’/βˆ’</sup></i> during <i>K. pneumonia</i> pulmonary tract infection.</p

    Impaired survival and bacterial defense in LysM-<i>Myd88<sup>βˆ’/βˆ’</sup></i> and Tie2-<i>Myd88<sup>βˆ’/βˆ’</sup></i> mice.

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    <p>Control, LysM-<i>Myd88<sup>βˆ’/βˆ’</sup></i> and Tie2-<i>Myd88<sup>βˆ’/βˆ’</sup></i> mice were intranasally infected with ∼6Γ—10<sup>3</sup> CFU <i>K. pneumoniae</i>. Survival of control (dark grey symbols, nβ€Š=β€Š37), LysM-<i>Myd88<sup>βˆ’/βˆ’</sup></i> (light grey symbols, nβ€Š=β€Š9) and Tie2-<i>Myd88<sup>βˆ’/βˆ’</sup></i> mice (white symbols, nβ€Š=β€Š13) expressed as Kaplan-Meier plot <b>(A)</b>, bacterial loads in lung <b>(B)</b>, blood <b>(C)</b>, spleen <b>(D)</b> and liver <b>(E)</b>, of control (dark grey bars, nβ€Š=β€Š8), LysM-<i>Myd88<sup>βˆ’/βˆ’</sup></i> (light grey bars, nβ€Š=β€Š8) and Tie2-<i>Myd88<sup>βˆ’/βˆ’</sup></i> mice (white bars, nβ€Š=β€Š5 mice). Data are expressed as box-and-whisker diagrams depicting the smallest observation, lower quartile, median, upper quartile, and largest observation. BC+β€Š=β€Šnumber of positive blood cultures. Survival curves were compared with Log-Rank test Bacterial loads were compared to control mice determined with Mann-Whitney <i>U</i> test: * <i>p</i><0.05, ** <i>p</i><0.01, *** <i>p</i><0.001.</p

    Genetic and functional characterization of primary cells from LysM-<i>Myd88<sup>βˆ’/βˆ’</sup></i> and Tie2-<i>Myd88<sup>βˆ’/βˆ’</sup></i> mice.

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    <p>The residual amount of the MyD88<sup>fl/fl</sup> allel in blood and primary cells LysM-<i>Myd88<sup>βˆ’/βˆ’</sup></i> and Tek-<i>Myd88<sup>βˆ’/βˆ’</sup></i> mice was quantified via qRT-PCR relative to the unaffected <i>Socs-3</i> gene. The amount of remaining β€œfloxed” MyD88 region in LysM-MyD88<sup>βˆ’/βˆ’</sup> and Tek-MyD88<sup>βˆ’/βˆ’</sup> mice was calculated using the 2<sup>-deltaCt</sup> (ΔΔCt) method using the amount of genomic DNA from <i>Myd88<sup>fl/fl</sup></i> mice for the no-deletion control. The deletion efficiency was calculated as (1 - residual Myd88<sup>fl</sup>) Γ—100% <b>(A)</b>. Whole blood <b>(B)</b>, alveolar and peritoneal macrophages <b>(C,D)</b> and splenocytes <b>(E)</b> derived from control, LysM-<i>Myd88<sup>βˆ’/βˆ’</sup></i> and Tie2-<i>Myd88<sup>βˆ’/βˆ’</sup></i> mice (nβ€Š=β€Š3 per group) were in vitro stimulated with LPS derived from <i>Klebsiella pneumoniae</i> (1 Β΅g/ml) or heat killed <i>K. pneumoniae</i> in two concentrations (2Γ—10<sup>5</sup> CFU/ml or 2Γ—10<sup>7</sup>/ml) for 20 hours. Data are expressed as mean (SE). * <i>p</i><0.05, ** <i>p</i><0.01, *** <i>p</i><0.001.</p

    Lung inflammatory response.

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    <p>Mice were intranasally infected with ∼6Γ—10<sup>3</sup> CFU <i>K. pneumoniae</i>; Histological scores 24 hours after infection determined as described in the Methods section, in control (dark grey, nβ€Š=β€Š8), LysM-<i>Myd88<sup>βˆ’/βˆ’</sup></i> (light grey, nβ€Š=β€Š8) and Tie2-<i>Myd88<sup>βˆ’/βˆ’</sup></i> mice (white, nβ€Š=β€Š5) <b>(A)</b>. Panel <b>(B)</b> shows representative lung histology of control, LysM-<i>Myd88<sup>βˆ’/βˆ’</sup></i> and Tie2-<i>Myd88<sup>βˆ’/βˆ’</sup></i> mice H&E staining, original magnification 20Γ—. Neutrophil influx compared between mouse groups as reflected by Ly6 lung surface positivity <b>(C)</b> and whole lung MPO levels <b>(D)</b>. Panel <b>(E)</b> shows representative images of Ly-6 staining on lung slides from control, LysM-<i>Myd88<sup>βˆ’/βˆ’</sup></i> and Tie2-<i>Myd88<sup>βˆ’/βˆ’</sup></i> mice; Data are expressed as box-and-whisker diagrams depicting the smallest observation, lower quartile, median, upper quartile, and largest observation. * <i>p</i><0.05, ** <i>p</i><0.01.</p

    The absence of MyD88 in the hematopoietic compartment determines the impaired antibacterial defense of Tie2-<i>Myd88<sup>βˆ’/βˆ’</sup></i> mice.

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    <p>Control and Tie2-<i>Myd88<sup>βˆ’/βˆ’</sup></i> mice were irradiated and injected with control or Tie2-<i>Myd88<sup>βˆ’/βˆ’</sup></i> bone marrow cells. Six weeks after transplantation, mice were infected with 6Γ—10<sup>3</sup> CFU <i>K. pneumoniae</i> and sacrificed after 24 hours. Bacterial loads in lung <b>(A)</b>, blood <b>(B)</b>, spleen <b>(C)</b> of control mice transplanted with control bone marrow (Co+ Co BM, grey bars, nβ€Š=β€Š8) and Tie2-<i>Myd88<sup>βˆ’/βˆ’</sup></i> mice transplanted with control bone marrow (Tie2-<i>Myd88<sup>βˆ’/βˆ’</sup></i>+Co BM, white dotted bars) or Tie2-<i>Myd88<sup>βˆ’/βˆ’</sup></i> bone marrow (Tie2-<i>Myd88<sup>βˆ’/βˆ’</sup></i>+Tie2-<i>Myd88<sup>βˆ’/βˆ’</sup></i> BM, white bars). Data are expressed as box-and-whisker diagrams depicting the smallest observation, lower quartile, median, upper quartile, and largest observation. * <i>p</i><0.05, ** <i>p</i><0.01.</p

    Bone marrow transfer restores responsiveness of hematopoietic cells from Tie2-<i>Myd88<sup>βˆ’/βˆ’</sup></i> mice to <i>Klebsiella</i>.

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    <p>Whole blood <b>(A)</b>, alveolar and peritoneal macrophages <b>(B,C)</b> and splenocytes <b>(D)</b> derived from control mice transplanted with control bone marrow (Co+ Co BM, grey bars) and Tie2-<i>Myd88<sup>βˆ’/βˆ’</sup></i> mice transplanted with control bone marrow (Tie2-<i>Myd88<sup>βˆ’/βˆ’</sup></i>+Co BM, white dotted bars) or Tie2-<i>Myd88<sup>βˆ’/βˆ’</sup></i> bone marrow (Tie2-<i>Myd88<sup>βˆ’/βˆ’</sup></i>+Tie2-<i>Myd88<sup>βˆ’/βˆ’</sup></i> BM, white bars). (nβ€Š=β€Š2–3 per group) were in vitro stimulated with LPS derived from <i>Klebsiella pneumoniae</i> (1 Β΅g/ml) or heat killed <i>K. pneumoniae</i> in two concentrations (2Γ—10<sup>5</sup> CFU/ml or 2Γ—10<sup>7</sup>/ml) for 20 hours. Data are expressed as mean (SE). * <i>p</i><0.05, ** <i>p</i><0.01. NDβ€Š=β€Šnot determined.</p
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