Figure 4

A) MDM were generated from CD14++ monocytes purified from PBMC by MACS separation followed by 5-day incubation with M-CSF (100 ng/ml). Cells remained untreated (none) or were stimulated with LPS (10 ng/ml) or with particle-mix of ultrafine P90 and fine TiO₂ (each with 32 μg/ml) for 3 h (n = 5 patients with COPD, n = 8 healthy controls, mean ± S.D.; * p < 0.05 compared to untreated controls). B) Effect of particles on CYP1B1 expression in sputum macrophages of healthy non-smokers, healthy smokers and patients with COPD. Sputum macrophages were purified using RosetteSep to deplete unwanted leukocytes. Cells remained untreated (none) or were stimulated with LPS (10 ng/ml) or with particle-mix of ultrafine P90 and fine TiO₂ (each with 32 μg/ml) for 3 h (n = 5 non-smokers, n = 4 smokers, n = 7 patients with COPD, mean ± S.D.; * p < 0.05). C) Cells remained untreated (none) or were stimulated with LPS (10 ng/ml), with particle mix of ultrafine P90 and fine TiO₂ (each with 32 μg/ml) or with each particle separately for 22 h (each with 32 μg/ml) (A549 n = 3, Calu-3 n = 4 experiments from different cell passages, mean ± S.D.; * p < 0.05 compared to untreated control). D) Effect of ultrafine P90 on CYP1B1 mRNA expressin in primary bronchial epithelial cells. Cells were obtained by bronchial brush biopsy. Cells remained untreated (none) or were stimulated with ultrafine P90 (32 μg/ml) for 3 h (n = 7 incubations from different donors, mean ± S.D.; * p < 0.05 compared to untreated control).