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

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

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

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

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