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. 2017 Apr 6;102(7):1161–1172. doi: 10.3324/haematol.2016.156869

Figure 6.

Figure 6.

ETB but nor ETA blockade prevents adhesion of neutrophils from healthy individuals and patients with sickle cell disease on activated endothelial cells and recombinant adhesion proteins. (A) Adhesion of neutrophils to endothelial cells (HMEC-1) over a 30 min period in a microfluidic system at a shear stress of 1 dyn/cm2, expressed as median area of neutrophil-associated fluorescence per field comparing six healthy controls (AA) to 12 SCD patients (SS). (B) Representative photomicrographs of anti-CD16 Alexa 488 (in green)-labeled neutrophils that have gradually adhered to the endothelial cells in microfluidic channels. Three channels were infused with the same batch of whole blood. Conditions were: NT: untreated blood; BQ123: blood preincubated with 1 μM BQ123; BQ788: blood previously treated with 1 μM BQ788. (C) Relative change in neutrophil adhesion on activated HMEC-1 after 30 min of infusion of whole blood from seven healthy AA controls (green circles) and 14 SS SCD patients (red triangles) previously incubated with 1 μM BQ123 or 1 μM BQ788 compared with non-treated blood. (D) Relative change in neutrophil adhesion 30 min after infusion of whole blood from five healthy donors (green circles) and five SCD patients (red triangles) on activated HMEC previously incubated with BQ123 or BQ788 compared with untreated HMEC. (E) Neutrophil adhesion on recombinant P-selectin, VCAM-1 and ICAM-1 proteins 30 min after infusion of whole blood from four healthy donors (green circles) and four SCD subjects (red triangles) previously incubated with BQ123 or BQ788 compared with non-treated blood. *P<0,05, ***P<0,001 with paired, non-parametric t-test (Wilcoxon test) vs. untreated blood or HMEC.