Figure 1.
Dose-dependent effects of rhesus θ-defensin (RTD)-1 on neutrophil migration in vitro. To assess RTD-1 effects, peripheral blood neutrophils were isolated and (A) 4 × 105 cells in Rosewell Park Memorial Institute medium were pretreated with RTD-1 or PBS for 15 minutes and placed in the apical chamber of their respective transwells. Inserts were then placed in the presence of 15 ng/ml CXCL8 to induce chemotaxis. Cells in the basolateral chamber are expressed as percent normalized migrated cells; n = 3 per group; mean (±SD). Horizontal dashed line indicates 50% migration. Treatment compared with control by one-way ANOVA. Neutrophil–endothelial cell adhesion was assessed by exposing endothelial cells to 100 ng/ml TNF before incubation with drug-treated neutrophil. Adhesion was assessed by measuring fluorescence from the calcein-AM–labeled neutrophils (B and C); n = 3 per group (technical replicates) taken from two experiments; mean (±SD). Representative images from a third experiment (C). Magnification ×10. Treatment compared with control by unpaired two samples t test. *P < 0.05, ***P < 0.001; HMVEC-L = human microvascular endothelial lung cells; PMN = polymorphonuclear leukocyte.