Xpb82d3a2

Mice were sensitized intranasally (i.n.) with 50 μg of HDM (Dermatophagoides. pteronyssinus; Greer Laboratories, XPB82D3A2.5) on day 1, followed by 10 μg of HDM (i.n.) on days 8 and 13. In some experiments, mice were treated with 3-MA (i.p., 300 mg; Sigma-Aldrich, M9281), dexamethasone (i.p., 1 mg/kg; Sigma-Aldrich, D4902) or phosphate-buffered saline (PBS; Gibco, 14,190), anti-mouse IL33 antibody (i.n., 6 μg/mice, clone 396,118; R&D, AF3626) or Goat IgG isotype controls (R&D, AB-108-C), and anti-mouse TSLP antibody (i.n., 20 μg/mice, clone 152,614; R&D, MAB555) or Rat IgG2A isotype control antibody (R&D, MAB006), one day before each HDM challenge. Two days after the last HDM challenge, mice were anesthetized using 300 μl i.p. injection of ketamine (10 mg/ml) and xylazine (1 mg/ml). Measurements of airway resistance and dynamic compliance were conducted using the Fine Pointe RC system (Buxco Research Systems), in which mice were mechanically ventilated using a modified version as described previously [18 (link),68 (link)]. Mice were sequentially challenged with aerosolized PBS (baseline), followed by increasing doses of methacholine (2.5 mg/ml, 5.0 mg/ml, 10 mg/ml, 20 mg/ml and 40 mg/ml); Sigma-Aldrich, A2251. R_L and C_dyn values were recorded during a 3-min period after each methacholine challenge.