Rolling on E-selectin or P-selectin evokes extended, but not high affinity LFA-1, in human neutrophils. (A) Whole blood was perfused through flow chambers coated with E-selectin and isotype antibody, KIM127, NKI-L16, mAb 24, or TS1-22 for 2 minutes at 5.94 dyn/cm2. (B) Isolated human neutrophils were stimulated with 1mM Mn2+ and immediately perfused through flow chambers coated with E-selectin and isotype antibody, mAb 24 or TS1-22 for 2 minutes at 5.94 dyn/cm2. (C) Whole blood was treated with piceatannol (200μM at 37°C for 60 minutes), PP2 (10μM at 37°C for 10 minutes), or SB203580 (20mM at 37°C for 30 minutes) before the perfusion through the flow chamber coated with E-selectin and isotype antibody or KIM127. (D) Whole blood was perfused through flow chambers coated with P-selectin and isotype antibody, KIM127, NKI-L16, mAb 24, or TS1-22 for 2 minutes at 5.94 dyn/cm2. (E) Purified human neutrophils were stimulated with 1mM Mn2+ and immediately perfused through flow chambers coated with P-selectin and isotype antibody, mAb 24, or TS1-22 for 2 minutes at 5.94 dyn/cm2. Number of neutrohils was counted after the Wright-Giemsa staining. Data are normalized to E-selectin or P-selectin and KIM127 (= 100%; panels A, C, and D) or E-selectin or P-selectin and TS1-22 (= 100%; panels B and E) and presented as mean ± SEM. *P < .05 from isotype control. †P < .05 from KIM127. Data are representative of at least 3 independent experiments.
