PI3K is involved in L-selectin- and PSGL-1-mediated neutrophil rolling on E-selectin via F-actin redistribution and assembly.

L-selectin and P-selectin glycoprotein ligand-1 (PSGL-1) are adhesion molecules that play critical roles in neutrophil rolling during inflammation and lymphocyte homing. On the other hand they also function as signaling receptors to induce cytoskeleton changes. The present study is to investigate the signaling kinases responsible for the F-actin changes mediated by L-selectin and PSGL-1 during neutrophil rolling on E-selectin. Western blot analysis demonstrated that PI3K activation, peaking within 5 min, was induced by ligation of L-selectin and PSGL-1 with E-selectin, and that Vav1 (the pivotal downstream effector of PI3K signaling pathway involved in cytoskeleton regulation) was recruited to the membrane and tyrosine-phosphorylated, depending on PI3K. Furthermore, the F-actin redistribution and assembly mediated by ligation with E-selectin were blocked by LY294002, a PI3K specific inhibitor. Additional experiments showed that PI3K activity was involved in neutrophil rolling on E-selectin. However, Syk/Zap70, the well-known upstream kinase of PI3K, was not involved in this event. These data suggest that PI3K is required for the F-actin-based cytoskeleton changes during neutrophil rolling on E-selectin, which may consequently regulate the rolling event.