Anti-TF antibodies suppress early eosinophil transmigration under flow conditions. Endothelial cells were treated with 20 ng/mL IL-4 for 24 hours. Stimulated endothelial cells were assembled into the flow chamber and freshly isolated eosinophils (5 × 105/mL) were perfused over the monolayer at 2 dyn/cm2. The endothelial cells and eosinophils were pretreated for 15 minutes with buffer alone, 10 μg/mL VIC7 or VD8. (A) Total accumulated cells were determined between 4 and 5 minutes as described in “Materials and methods.” Over 90% of the accumulating cells were firmly adherent. No statistically significant differences were observed between the mean values. (B) Transmigration was assessed from multiple fields between 3 and 4 minutes or (C) between 6 and 7 minutes. (D) Transmigration kinetics as assessed every 15 seconds between 2 and 5 minutes. In some cases, the endothelial cells were selectively precoated with VD8 (VD8(EC)). Data represent means ± SEM of at least 3 experiments. *P < .05 (versus control).
