Administration of anti-IL-22 neutralizing antibody enhances BLM-induced pulmonary inflammation and fibrosis. Mice were induced with once bleomycin (BLM) in combination with intraperitoneal injection of anti-IL-22 neutralizing antibody (Anti-IL-22 Ab) or isotype antibody (Isotype Ab) for two consecutive weeks, as compared with BLM-treated mice (BLM control) and saline-treated mice (saline control) without any administration followed. (a) Cell counts of lymphocytes from bronchoalveolar lavage fluid (BALF) were shown. (b) Representative photomicrographs of lung sections were stained with hematoxylin and eosin (H&E) or with Masson’s trichrome, as well as by immunohistochemistry for anti-α-smooth muscle actin (α-SMA), anti-collagen I (Col I), anti-collagen III (Col III), and anti-transforming growth factor (TGF)-β antibody. μm (for H&E and Masson’s trichrome staining), or 50 μm (for immunohistochemistry). (c) Levels of transcripts for -sma, col1a2, col3a1, and tgf-β in the lungs were measured by real-time reverse transcription-polymeras chain reaction (RT-PCR) analysis, normalized to that of saline-treated mice. (d) Level of transcript for tgf-β in the lungs was assessed as described in Figure 4(c) (left). Protein levels of phosphorylated Smad2 (pSmad2), total Smad2, and GADPH in lung homogenates were determined by western blotting (middle). Densitometry was performed, and fold increase in pSmad2 expression after normalization to total Smad2 expression was shown (right). Values in (b), (c), and (d) are the mean and SEM of 5 to 9 mice per group from at least two separate experiments. ; .
