FIGURE 5.

Lipoxin B₄ enhances the lymphatic transmigration of neutrophils from patients with atherosclerosis but does not affect neutrophils from healthy controls. Neutrophils were isolated from healthy controls or patients with atherosclerosis and added to the upper well of a transwell with a confluent lymphatic endothelial cell monolayer. The neutrophils transmigrated for three hours, after which the cells in the lower well were counted to determine the migration rate. The migration experiment was conducted using either vehicle or the chemoattractant interleukin (IL)‐8 (5 ng/ml) in the lower well. (A) A schematic illustration of the transmigration assay is shown. (B) The percentage of neutrophils that migrated across lymphatic endothelial cell monolayer were quantified, either spontaneously or in response to the chemoattractant IL‐8. The migration pattern was investigated in neutrophils isolated from healthy controls (n = 6, white bars) and patients with atherosclerosis (n = 6, gray bars). (C) Neutrophils were treated with vehicle (0.01% ethanol), lipoxin A₄ (LXA₄:1 nM, blue bars), or lipoxin B₄ (LXB₄: 1 nM, green bars) for 15 minutes prior to the transmigration assay. Lipoxin‐induced changes to the neutrophil migration rate were calculated as the log₂ fold change relative to respective vehicle‐treated condition. Assuming non‐Gaussian distribution of the human samples, statistical analysis was determined using Mann–Whitney U test when comparing two groups, and Kruskal–Wallis test with Dunn's post hoc comparisons when comparing over two groups. Data are presented as mean ± standard error of the mean (SEM). Statistical significance is indicated as *p < .05, **p < .01