23 research outputs found

    IL-37 dampens inflammasome activation in phagocytic cells.

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    <p>Alveolar macrophages and epithelial cells from naive mice and peripheral neutrophils were pre-exposed to recombinant IL-37 precursor for 8 hours before stimulation with live <i>Aspergillus</i> conidia for 2 hours. (<b>A</b>) Percent of phagocytosis and conidiocidal activity. (<b>B</b>) <i>Il1b</i> and <i>Nos2</i> mRNA expression by RT-PCR on total lung cells. Ct, control cells. None, <i>Aspergillus</i>-pulsed, untreated cells. (<b>C</b>) Activation of distinct intracellular kinases in RAW cells, using Proteome Profiler Array, pre-exposed to 100 ng/ml IL-37 for 8 hours before stimulation with live <i>Aspergillus</i> conidia for 30 min. Data are representative (Proteome Profiler Array) or pooled from two experiments. *P<0.05,**P<0.01, ***P<0.001, IL-37-stimulated <i>vs</i> unstimulated cells.</p

    NLRP3-deficient mice exhibit reduced neutrophil recruitment and IL-1β production in pulmonary aspergillosis.

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    <p>C57BL/6 and <i>Nlrp3<sup>−/−</sup></i> mice were infected intranasally with <i>A. fumigatus</i> and treated with 1000 ng/mouse recombinant IL-37 precursor administered intraperitoneally 1 hour before the infection. (<b>A</b>) BAL fluid morphometry [number of total (T) cells and polymorphonuclear neutrophils (P). Values represent the mean±SD of three mice per group and are representative of 3 independent experiments]. (<b>B</b>) Lung histology (periodic acid-Schiff staining) and cell recruitment (insets). Scale bars, 100 µm and 25 µm, respectively. Arrows indicate neutrophils. (<b>C</b>) <i>Mpo</i> and <i>Cxcl2</i> mRNA expression by RT-PCR on total lung cells. (<b>D</b>) IL-1β production (ELISA on lung homogenates). Assays were done 3 days after the infection. Data are representative (histology) or pooled from three experiments. *P<0.05, <i>Nlrp3<sup>−/−</sup> vs</i> C57BL/6 mice and treated <i>vs</i> untreated (None) mice. Naïve, uninfected and untreated mice.</p

    IL-37 reduces inflammatory cell recruitment in mice with inflammatory aspergillosis.

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    <p>C57BL/6 mice were infected intranasally (in) with <i>A. fumigatus</i> and pretreated one time with different doses IL-37 administered intraperitoneally (ip) at different times before the infection. Mice were assessed for: (<b>A</b>) fungal growth (Log<sub>10</sub> CFU, mean±SD) in the lungs at 1 and 3 days post-infection (dpi); (<b>B</b>) BAL fluid morphometry [number of total (T) cells and polymorphonuclear neutrophils (P) upon May Grunwald Giemsa staining. Values represent the mean±SD of three mice per group and are representative of 3 independent experiments]; (<b>C</b>) lung histology (periodic acid-Schiff and, in the inset, TUNEL staining). Red arrows indicate PMN and white arrows indicate increased deposition of DNA on lung parenchyma (in TUNEL-stained sections). Scale bars, 25 µm; (<b>D</b>) myeloperoxidase (<i>Mpo</i>), <i>Cxcl1</i> and <i>Cxcl2</i> mRNA expression by RT-PCR on total lung cells; (<b>E</b>) lung histology (periodic acid-Schiff staining, scale bars, 25 µm) and (<b>F</b>) <i>Mpo</i>, <i>Cxcl1</i> and <i>Cxcl2</i> mRNA expression (RT-PCR on total lung cells) in mice pretreated with IL-37 given ip or in, at different hours before the infection. (<b>G</b>) Numbers of CD11b/Gr1–positive cells were assessed by flow cytometry of total lung cells from LPS-treated mice. Data are representative (histology) or pooled from three experiments. *P<0.05,**P<0.01, treated <i>vs</i> untreated (None) mice. Naïve, uninfected and untreated mice.</p

    IL-37 restrains inflammation in fungal allergy and <i>Cftr<sup>−/−</sup></i> mice.

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    <p>(<b>A</b>) Lung histology (PAS- and Masson's trichrome-stained sections, scale bars 100 and 25 (insets) µm); (<b>B</b>) hydroxyproline content (µg/lung); (<b>C</b>) expression of mucins (RT-PCR on total lung cells); (<b>D</b>) expression of cytokines and Th transcription factors in total lung cells from mice with ABPA and treated with IL-37. None, untreated mice. Naïve, uninfected and untreated mice. (<b>E</b>) Fungal growth (Log<sub>10</sub> CFU, mean±SD) in the lungs of <i>Cftr<sup>−/−</sup></i> mice infected intranasally with <i>A. fumigatus</i> and treated intraperitoneally with IL-37, at the dose of 1000 ng/mouse, 1 hour before the infection. (<b>F</b>) BAL fluid morphometry [number of total (T) cells and polymorphonuclear neutrophils (P) upon May Grunwald Giemsa staining]. (<b>G</b>) Lung histology (periodic acid-Schiff staining) and cell recruitment (insets). Scale bars, 100 µm and 25 µm in the insets; (<b>H</b>) <i>Mpo</i> and <i>Cxcl2</i> mRNA expression and (<b>I</b>) cytokine gene expression on total lung cells by RT-PCR, 3 days after the infection. Data are pooled from two experiments. *P<0.05, **P<0.01, treated <i>vs</i> untreated (None) mice. Naïve, uninfected and untreated mice.</p

    Surface analysis of resting conidia of <i>ags</i>Δ_<i>5T</i> mutant and parental (<i>ku80</i>) strains.

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    <p>(A): height images (z-range = 1 µm; recorded in water with silicon nitride tips). Atomic Force Microscopy (AFM) images showing the amorphous surface without the rodlet layer on the triple <i>ags</i>Δ_<i>5T</i> mutant conidia whereas the rodlet are observed on the parental strain conidial surface. (B): TEM observations. Note the presence of an extracellular material on the surface of the <i>ags</i>Δ_<i>5T</i> conidia (arrow); CW: cell wall. (C): SDS-PAGE (15% gel) of Hydrofluoric acid (HF) extracts of rodlets from resting conidia showing the two bands, 16 kDa and 14.5 kDa of RodAp classically seen from HF treatment of the conidia <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1003716#ppat.1003716-Aimanianda1" target="_blank">[10]</a>. Data are representative of at least three independent experiments.</p

    Cyclophosphamide immunosuppressed mice and anti-Ly6G treated neutropenic mice infected with resting conidia of <i>ags</i>Δ_<i>5T</i> and parental (<i>ku80</i>) strains.

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    <p>(A–C) Cyclophosphamide immunosuppressed mice; (D–E) anti-Ly6G treated neutropenic mice; (A) Survival (%) and (B) fungal growth estimated as CFUs in lung. (C and E) lung histology (periodic acid-Schiff-staining). Note the polymorphonuclear cells and mononuclear infiltrates surrounding the bronchi in <i>ku80</i> infected lung. (D) Histological appearance of lungs of anti-Ly6G neutropenic mice infected with conidia of <i>ags</i>Δ_<i>5T</i> and <i>ku80</i> (Gomori's methanamine silver-staining). Note the absence of mycelial development of <i>ags</i>Δ_<i>5T</i> conidia in neutropenic mice. Data are representative of at least three independent experiments. *: p<0.05.</p

    Working model explaining sequential and differential immune events upon inhalation of the <i>ags</i>Δ mutant and the parental (<i>ku80</i>) strain conidia.

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    <p>The presence of the glycoprotein layer on the triple <i>ags</i>Δ mutant conidial surface hides the rodlet layer. Increased exposure of PAMPs (WGA and ConA positive molecules and β-(1,3)-glucans) during vegetative growth in the triple <i>ags</i>Δ mutant modifies the host immunological response. This facilitates phagocytosis and killing of the triple <i>ags</i>Δ mutant and stimulates pro-inflammatory immune responses.</p
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