Patient neutrophil dysfunction. (A) (Left column) Temporal changes in forward angle light scatter (polarization) in normal neutrophils and neutrophils from patients I.2.2 and I.2.3 after treatment with buffer or fMLF (1 × 10−7 M). (Right column) Similar time course of changes in right angle light scatter (membrane ruffling). In B, neutrophils (1 μL of 2 × 106 cells/mL in HBSS with divalent cations) were added to the “Cell” well of EZ-TAXIScan and either buffer or fMLF was added to the “Chemoattractant” well. The cells were incubated for 60 minutes, and images were collected every 2.5 minutes. Ten randomly chosen cells were electronically traced using the acquired images and the paths of the cells plotted with the position at t = 0 anchored at the origin. (Left column) Random migration of neutrophils from a normal subject and from patients I.2.2 and I.2.3. (Right column) Directed migration in response to fMLF (1 × 10−8 M). (C) Scattergrams of the average velocities of the individual cells that were tracked in B. Note that the cells from both patients have a significant defect (analysis of variance, ****P < .001) in both their basal migration and a defect in their migration to the chemoattractant, fMLF.
