Figure 3

CSE-induced adhesion of PMNs to fibrinogen is inhibited after pre-incubation with α-CD11b and α-CD18. Calcein-AM labeled PMNs were added to fibrinogen-coated wells and incubated with indicated reagents for 30 minutes at 37°C. After incubation, fluorescence was measured before and after three washes with HEPES incubation buffer. (A) Cigarette smoke extract (CSE; OD 0.06-0.24) induces adhesion of PMNs to fibrinogen (***, p < 0.001 CSE vs. control. Legend: each symbol represents a different donor (N = 8, n = 3-8). (B) After labeling, PMNs were pre-incubated for 20 minutes with medium (white bars), α-CD18 (10 μg/ml; grey bars) or α-CD11b (10 μg/ml; black bars) at 37°C prior to incubation with medium or cigarette smoke extract (CSE) for 30 minutes at 37°C. Cigarette smoke extract (CSE; OD 0.06-0.24) induces adhesion of PMNs to fibrinogen (^, p < 0.05; ^^, p < 0.01 CSE vs. control), which is blocked by α-CD18 and α-CD11b (*, p < 0.05; **, p < 0.01 CSE α-CD11b/CD18 vs. CSE no Ab). FMLP (10^-8 M) induces adhesion of PMNs to fibrinogen (⁺⁺, p < 0.01 fMLP vs. control), which is blocked by α-CD18 and α-CD11b (*, p < 0.05; **, p < 0.01 fMLP α-CD11b/CD18 vs. fMLP no Ab). Legend: each symbol represents a different donor (N = 4, n = 3).