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To perform bronchoalveolar lavage (BAL), a 20-gauge angiocath ligated into the trachea and 1 ml of sterile PBS was instilled, then removed through the angiocath; the process was repeated three times. The samples were pooled and a 200 μl aliquot of the BAL fluid was placed in a cytospin and centrifuged at 500 g for 10 min. The pellet was re-suspended in 1 ml of PBS, plated on glass slides, stained with crystal violet and subjected to a blinded manual cell count and differential. Cytokine and chemokine levels in the supernatant were measured using the BD Cytometric Bead Array (BD Biosciences). Samples were analysed in triplicate using the Mouse Inflammation Kit (BD Biosciences) according to the instructions provided. This kit detects interleukin (IL)-6, MCP-1 and tumour necrosis factor (TNF)-α. Total transforming growth factor (TGF)-β1 was measured using the TGF-β1 RII DuoSet enzyme-linked immunosorbent assay (ELISA, R&D Systems, Minneapolis, MN) according to the manufacturer’s instructions. Total interleukin (IL)-1β was measured using the IL-β1 Ready-Set-GO ELISA (Ebioscience, San Diego, CA) according to the manufacturer’s instructions. Caspase-1 activity was measured using the Caspase-1 Fluorometric Assay (R&D Systems), according to the manufacturer’s instructions.