Figure 5
TNFα release is diminished in pulmonary neutrophils from PAR-2-deficient mice compared to wild type mice. A. Whole lungs from naïve BALB/c and PAR-2 mice were dissociated and neutrophils were isolated by Percoll gradient. Neutrophils (1 × 106) were then treated ex vivo with PBS or GC frass and 18 h later, the supernatants were harvested, clarified, and analyzed for TNFα levels by ELISA. Data are expressed as means ± SEM (n = 3 separate experiments run in duplicate) and statistical significance was analyzed by ANOVA (*p < 0.001). B. BALB/c and PAR-2-deficient mice were administered a single intratracheal instillation of GC frass and 18 h later BAL fluid was harvested. Neutrophils (5 × 105) were isolated by Percoll gradient and were cultured without additional treatment for 6 h. Supernatants were then harvested, clarified and analyzed for TNFα levels by ELISA. Data are expressed as means ± SEM (n = 5 mice per group) and statistical significance was analyzed by Student’s t-test (*p = 0.002).