Characterization of the airway inflammatory response in the chronic model of HP: Histology and cytokine.

<p>WT and <i>Cd103</i>^<i>-/-</i> mice were exposed to SR for 3 weeks and a section of the left lobe was used for histology while other lobes were used for analysis of cytokine production. <b>A)</b> Lung sections of WT and <i>Cd103</i>^<i>-/-</i> mice exposed to saline or SR. <b>B)</b> Histological score was obtained and compared between WT and <i>Cd103</i>^<i>-/-</i> mice. <b>C)</b> Flow cytometry gating strategy for the polarity of the effector lung response after <i>ex vivo</i> stimulation of lung leukocytes isolated from SR-exposed mice. CD4⁺ T cells were gated from total lung cells and cytokine-positive cells were analyzed using Fluorescence Minus One (FMO) controls. <b>D)</b> Number of cells and <b>E)</b> % of IFNg⁺, IL-13⁺ and IL-17A⁺CD4⁺ T cells in the lung of saline and SR-exposed mice. Results representative of 2 different experiments; n = 4–6 mice/group. * = p < 0.05.</p

Chorographie du Royaume de Léon des Provinces des Asturies et de Galice

Representación planimétrica de hidrografía, vales, pontes, vías de comunicacion, abadías, castelos e bispadosLiñas límite de reinos, iluminadas con técnica de aguadaO relevo está representado por perfís abatidos (pequenas montañas)Toponimia local en castelán aínda que con erros. Cartela, rosa dos ventos e escalas gráficas en francésOrixe de lonxitude na illa del HierroContén rosa dos ventos indicando a orientación, expresando cunha flor de lis a direccion N

Transfers of DCs and CD4⁺ T cells.

<p>WT and <i>Cd103</i>^<i>-/-</i> DCs were injected in a criss-cross manner in WT and <i>Cd103</i>^<i>-/-</i> mice. 24h after injection, recipient mice were submitted to either the acute or chronic model. <b>A)</b> Total BAL neutrophils (cells/mL) and <b>B)</b> total BAL cells (cells/mL) of SR-exposed mice in the acute model. <b>C)</b> Total BAL lymphocytes (cells/mL) of SR-exposed mice in the chronic model. WT or <i>Cd103</i>^<i>-/-</i> CD4⁺ T cells were injected in <i>Rag</i>^<i>-/-</i> mice. 1 month after the transfer, recipient mice were submitted to the chronic model. <b>D)</b> Total BAL cells (cells/mL) and <b>E)</b> total BAL lymphocytes (cells/mL) of saline and SR-exposed mice. Results were pooled from two experiments; n = 6–10 mice/group. * = p < 0.05 with multi-comparison test.</p

Modulation of CD103 expression on DCs in response to SR <i>in vitro</i>.

<p>DCs were isolated from lung and spleen of WT mice and stimulated with 0μg/mL (ctrl), 1μg/mL or 5μg/mL of SR extract for 18h. DCs were identified as autofluorescence^-/ CD11c⁺/ MHC-II^hi cells and CD103 and XCR1 expression was analyzed as shown in <b>A)</b>. <b>B)</b> Flow cytometry examples of CD103 and XCR1 expression on SR-stimulated spleen- and lung-isolated DCs. <b>C)</b> Viability, measured with trypan blue, of spleen- and lung-isolated DCs after stimulation with SR. <b>D)</b> % of spleen- and lung-isolated DCs expressing CD103 (CD103⁺XCR1^-) or XCR1 and CD103 (CD103⁺XCR1⁺). <b>E)</b> Mean Fluorescence Intensity (MFI) of CD103 from spleen- and lung-isolated CD103⁺XCR1⁺ DCs. <b>F)</b> Transcript level of CD103 relative to Rplp0 and GNB in spleen- and lung-isolated DCs. <b>G)</b> <i>In vivo</i> CD103⁺ or CD103⁺XCR1⁺ on lung CD11c⁺/ MHC-II^hi DCs (as analyzed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0179678#pone.0179678.g004" target="_blank">Fig 4A</a>) 18h after SR exposure. Results are representative of 2 separate experiments; n = 5–6 mice/group. * = p < 0.05. † = p < 0.05 with multi-comparison test.</p

Airway inflammatory response in WT and <i>Cd103</i>^<i>-/-</i> mice in the acute model of HP.

<p>WT and <i>Cd103</i>^<i>-/-</i> mice were exposed to SR for 3 days and the BAL content was evaluated: <b>A)</b> BAL total and differential cell numbers (cells/mL) in saline and SR-exposed mice; <b>B)</b> BAL differential cell % (MΦ: macrophages, Ly: lymphocytes, Ne: neutrophils, Eo: eosinophils) in saline and SR-exposed mice. C-E): WT and <i>Cd103</i>^<i>-/-</i> mice were exposed to SR and the BAL content was evaluated 2h, 12h and 18h after the exposure: <b>C)</b> BAL total numbers (cells/mL) in saline and SR-exposed mice; <b>D)</b> BAL differential numbers (cells/mL) and <b>E)</b> % 18h after exposure to saline or SR. For A and B, results from 2 experiments were pooled; n = 6–12 mice/group. For C and D, results are representative of 2 different experiments; n = 3–4 mice/group. * = p < 0.05.</p

Characterization of the airway inflammatory response in the chronic model of HP: BAL and IgGs.

<p><b>A)</b> Timeline of the chronic and acute models of exposure to SR. Full line represents days of intranasal instillation of either saline or SR while dashed line represents day of euthanasia. B-E): WT and <i>Cd103</i>^<i>-/-</i> mice were exposed to SR for 3 weeks and the BAL content and serum immunoglobulins were measured. <b>B)</b> BAL total and differential cell numbers (cells/mL) in saline and SR-exposed mice. <b>C)</b> BAL differential cell % (MΦ: macrophages, Ly: lymphocytes, Ne: neutrophils) in saline and SR-exposed mice. SR-specific serum levels of <b>D)</b> IgG₁ and <b>E)</b> IgG_2a. For B and C, results were pooled between two experiments, and are representative of over 5 separate experiments; n = 8–12 mice/group. For D and E, results are representative of 2 different experiments; n = 4–6 mice/group. * = p < 0.05.</p

Human and Mouse Eosinophils Differ in Their Ability to Biosynthesize Eicosanoids, Docosanoids, the Endocannabinoid 2-Arachidonoyl-glycerol and Its Congeners

High eosinophil (EOS) counts are a key feature of eosinophilic asthma. EOS notably affect asthmatic response by generating several lipid mediators. Mice have been utilized in hopes of defining new pharmacological targets to treat asthma. However, many pinpointed targets in mice did not translate into clinics, underscoring that key differences exist between the two species. In this study, we compared the ability of human (h) and mouse (m) EOS to biosynthesize key bioactive lipids derived from arachidonic acid (AA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). hEOS were isolated from the blood of healthy subjects and mild asthmatics, while mEOSs were differentiated from the bone marrow. EOSs were treated with fatty acids and lipid mediator biosynthesis assessed by LC-MS/MS. We found that hEOS biosynthesized leukotriene (LT) C4 and LTB4 in a 5:1 ratio while mEOS almost exclusively biosynthesized LTB4. The biosynthesis of the 15-lipoxygenase (LO) metabolites 15-HETE and 12-HETE also differed, with a 15-HETE:12-HETE ratio of 6.3 for hEOS and 0.727 for mEOS. EOS biosynthesized some specialized pro-resolving mediators, and the levels from mEOS were 9-times higher than those of hEOS. In contrast, hEOS produced important amounts of the endocannabinoid 2-arachidonoyl-glycerol (2-AG) and its congeners from EPA and DHA, a biosynthetic pathway that was up to ~100-fold less prominent in mEOS. Our data show that hEOS and mEOS biosynthesize the same lipid mediators but in different amounts. Compared to asthmatics, mouse models likely have an amplified involvement of LTB4 and specialized pro-resolving mediators and a diminished impact of the endocannabinoid 2-arachidonoyl-glycerol and its congeners.Other UBCNon UBCReviewedFacultyResearche

Spatiotemporal signaling underlies progressive vascular rarefaction in myocardial infarction

Abstract Therapeutic angiogenesis represents a promising avenue to revascularize the ischemic heart. Its limited success is partly due to our poor understanding of the cardiac stroma, specifically mural cells, and their response to ischemic injury. Here, we combine single-cell and positional transcriptomics to assess the behavior of mural cells within the healing heart. In response to myocardial infarction, mural cells adopt an altered state closely associated with the infarct and retain a distinct lineage from fibroblasts. This response is concurrent with vascular rarefaction and reduced vascular coverage by mural cells. Positional transcriptomics reveals that the infarcted heart is governed by regional-dependent and temporally regulated programs. While the remote zone acts as an important source of pro-angiogenic signals, the infarct zone is accentuated by chronic activation of anti-angiogenic, pro-fibrotic, and inflammatory cues. Together, our work unveils the spatiotemporal programs underlying cardiac repair and establishes an association between vascular deterioration and mural cell dysfunction