Figure 3

(A) Proportion of small sputum macrophages after LPS inhalation. Sputum induction was performed in healthy control donors 24 h before (left plot) and 24 h after (right plot) inhalation of 20 µg LPS. Sputum macrophages were purified by RosetteSep and regated in a forward versus side scatter plot after staining with CD66b/CD16b/CD14. Percentage of small sputum macrophages before LPS inhalation is 13.4% and 66.5% 24 h after inhalation. Shown is 1 out of 4 donors. The average percentage of small sputum macrophages was 18.3% ± 19.3% before and 63.7% ± 25.7 % after LPS inhalation (P < 0.05). (B) Chemokine expression before and after LPS inhalation. Macrophages from induced sputum were purified 24 h before and 24 h after LPS inhalation. RT-PCR was performed for CCL2, CCL7, CCL13 and CCL22. mRNA expression before LPS inhalation (left bars) was set as one and shown is the fold difference after LPS challenge (right bars). This was 6.5 ± 4.5 for CCL2, 3.2 ± 1.2 for CCL7, 0.3 ± 0.1 for CCL13 and 0.9 ± 0.4 for CCL22. Shown are the mean values from four donors. *P < 0.05 **P < 0.01 ***P < 0.001.