Phenotype profile CD68⁺ F4/80⁻ cells in the lungs following pancreatitis.

<p>Flow cytometry analysis of CCR2, CD11c (M1) and CD206 (M2) activation markers of lung macrophages gated for FSC/SSC (R1) and CD68⁺F4/80⁻. Significant enrichment of the number of CCR2⁺ CD11c⁻ (A) and CD206⁺ CD11c⁻ (B) cells in the lungs following pancreatitis (ligated) compared to sham controls. Bars show mean ±SEM, n = 8 per group. *<i>P</i><0.05 versus sham control, by two-tailed Student <i>t</i>-test.</p

Acute pancreatitis induces changes in local pancreatic and lung chemokine levels.

<p>Tissue homogenates from pancreas (A, C) and lungs (B, D) were evaluated by ELISA for CXCL1 (A, B) and CCL2 (C, D) levels at the indicated time points. Both CXCL1 and CCL2 levels were significantly elevated in the pancreas (A) and lungs (B) 9, 24 and 48 h after pancreatitis induction compared to sham controls. Bars show mean ± SEM, n = 10 per group. *<i>P</i><0.05, **<i>P</i><0.01, ***<i>P</i><0.001 versus sham control, by two-tailed Student <i>t</i>-test.</p

Changes in pancreatic and lung neutrophil number during acute pancreatitis.

<p>Representative photomicrographs of pancreas (A) and lung (C) tissue stained for NIMP-R14, showing infiltration of neutrophils (brown) 24 h after induction of pancreatitis (original magnification, ×20). The number of NIMP-R14⁺ neutrophils in the pancreas (B) and lungs (D) were significantly enriched following pancreatitis induction (ligated) compared to sham control. Lung MPO activity was significantly increased following acute pancreatitis compared to sham controls at all time points evaluated (Fig. E). Bars show mean ± SEM, n = 10 per group. *<i>P</i><0.05, **<i>P</i><0.01, ***<i>P</i><0.001 versus sham control, by two-tailed Student <i>t</i>-test.</p

Changes in lung macrophage sub-populations during acute pancreatitis.

<p>Single cell preparations of the right lung were evaluated by flow cytometry. Dot plots from one representative experiment of sham control (A) and 24 h post pancreatitis induction (B) showing the gating strategy. Significant modulations in the percentage of R1 (C) and R2 (D) gated populations following acute pancreatitis compared to sham operated animals. Representative profiles of CD68 and F4/80 expressing cells in the R1 population of sham (D) and ligated (E) mice after 24 h are shown. A significant enrichment in the total number of R1 gated CD68⁺ F4/80⁻ cells in the right lung 9 h (F) and 24 h (G) after pancreatitis induction compared to sham controls. CD68⁺ cells were increased significantly in the immunohistochemical staining of the lung sections in the acute pancreatitis compared to sham at 9, 24 and 48 h. , n = 8 per group. *<i>P</i><0.05, **<i>P</i><0.01, ***<i>P</i><0.001 versus control, by two-tailed Student <i>t</i>-test.</p

Changes in pancreatic and lung F4/80⁺ macrophage number during acute pancreatitis.

<p>Representative photomicrographs of pancreas (A) and lung (C) sections stained for F4/80 as indicator of macrophage infiltration (brown) 24 h after pancreatitis induction (original magnification, ×20). Quantification of F4/80⁺ cells showed a significantly increased enrichment in the pancreas from 9 h following pancreatitis compared to sham controls (B). No significant difference was observed in the number of lung F4/80⁺ cells between pancreatitis and sham control groups (D). Bars show mean ± SEM, n = 10 per group. **<i>P</i><0.01, ***<i>P</i><0.001, by two-tailed Student <i>t</i>-test.</p

Morphological and physiological changes associated with acute pancreatitis and associated acute lung injury.

<p>Representative H&E images of pancreas (A, B) and lungs (C, D) 24 hours after induction of pancreatitis through BPD ligation (A, C) are depicted. Pronounced edema and necrosis observed in the pancreas after pancreatitis induction (B) compared to sham control (A). Lung section shows thickening of alveolar septa, hemorrhage and edema after pancreatitis induction (D) compared to sham control (C). Original magnification, 20×. Mice experienced a significant weight loss 24 and 48 h after induction of pancreatitis through BPD ligation compared to sham control (E). Data expressed as mean ± SEM, n = 10 per group. *<i>P</i><0.05, ***<i>P</i><0.001 versus sham control, by two-tailed Student <i>t</i>-test.</p

Additional file 3: Figure S2. of IL-1β mediates lung neutrophilia and IL-33 expression in a mouse model of viral-induced asthma exacerbation

Increased immune cells and total protein in BALF in both WT and IL-1β−/− groups 24 h after the last HDM challenge alone. Total protein in BALF (a), total cells from BALF (b) and total neutrophil count in BALF (c). Data are presented as mean ± SEM, n = 4–7 mice in each group. ## = p < 0.01 compared to respective HDM/Saline control, and # = p < 0.05 compared to respective HDM/Saline control. (TIFF 941 kb

Additional file 4: Figure S3. of IL-1β mediates lung neutrophilia and IL-33 expression in a mouse model of viral-induced asthma exacerbation

Increased immune cell count and chemokine expression at exacerbation, although in both WT and IL-1β−/− mice. Total eosinophil count (a), macrophage count (b) and lymphocyte count (c) from BALF at exacerbation. Chemokine mRNA expression of CCL11 (d) and CCL2 (e) from lung tissue homogenates were analysed with RT-qPCR. The relative gene expression was related to the reference gene 18S and normalised to control. Data are presented as mean ± SEM, n = 4–8 mice in each group. ## = p < 0.01 compared to respective HDM/Saline control. (TIFF 1200 kb

Additional file 2: Table S1. of IL-1β mediates lung neutrophilia and IL-33 expression in a mouse model of viral-induced asthma exacerbation

Commercially available primer sequences for genes analysed with RT-qPCR. (DOCX 13 kb