NEUTROPHILS USE BOTH SHARED AND DISTINCT MECHANISMS TO ADHERE TO SELECTINS UNDER STATIC AND FLOW CONDITIONS

Both P-selectin glycoprotein ligand-1 (PSGL-1) and L-selectin are localized on the microvilli of neutrophils and have been implicated in the attachment of neutrophils to P-selectin or E-selectin, We directly compared the attachment and rolling of neutrophils on P-selectin and E-selectin under flow, with emphasis on the functions of PSGL-1 and L-selectin, Flowing neutrophils attached more avidly and roiled at lower velocities on P-selectin than on E-selectin at matched densities, Studies with purified molecules indicated that P-selectin and E-selectin bound to a related site on PSGL-1 that overlapped the epitope for the anti-PSGL-1 mAb PL1, E-selectin bound with lower affinity than P-selectin to this site and also bound to an additional site(s) on PSGL-1, PL1 abolished adhesion of neutrophils to P-selectin under shear or static conditions, whereas DREG-56, a mAb to L-selectin, had no effect on adhesion to P-selectin, PL1 inhibited attachment of neutrophils to E-selectin under flow but not static conditions, DREG-56 also inhibited attachment of flowing neutrophils to E-selectin, and a combination of DREG-56 and PL1 nearly eliminated attachment to E-selectin under flow, These data suggest that PSGL-1 functions cooperatively with L-selectin to mediate optimal attachment of flowing neutrophils to E-selectin but not to P-selectin. Neutrophils attach more efficiently and with greater strength to P-selectin, perhaps because of the higher affinity of P-selectin for the PL1-defined site on PSGL-1.