Redistribution of adhesion receptors to the uropod of neutrophils stimulated by chemoattractants.

(i) Neutrophils were allowed to bind to coverslips coated with 20 μg/mL of an 80-kd fibronectin (FN80) fragment for 10 minutes at 37°C and stimulated with interleukin 8 (IL-8) for an additional 5 minutes. Neutrophils were then fixed and stained with the following monoclonal antibodies (mAb): anti-intercellular adhesion molecule 3 (ICAM-3) TP1/25 (B), anti-P-selectin glycoprotein ligand 1 (PSGL-1) KPL-1 (C), anti-CD43 TP1/36 (D), anti-CD44 HP2/9 (E), and anti-CD45 D3/9 (F). The control was unstimulated neutrophils stained with the anti-ICAM-3 TP1/25 mAb (A). (ii) Neutrophils were stimulated in the presence of 2 nmol/L of C5a (A, B, and C),N-formyl-methionyl-leucyl-phenylalanine (D, E, and F), or tumor necrosis factor (TNF-α) (G, H, and I) for 5 minutes at 37°C. Cells were then fixed and stained with the following mAb: anti-ICAM-3 TP1/25 (A, D, and G), anti-PSGL-1 KPL1 (B, E, and H), and anti-CD45 D3/9 (C, F, and I).