extravasation in vivo integrin is pivotal for integrin activation to bind ICAMs and leukocyte -chain phosphorylation of the human leukocyte CD11b/CD18 (Mac-1) α

Abstract The promiscuous CD11b/CD18 (Mac-1) integrin has important roles in regulating many immunological functions like leukocyte adhesion and emigration from the bloodstream via interactions with the endothelial ligands ICAM-1 and ICAM-2 , iC3b-mediated phagocytosis and apoptosis. However, the mechanisms for Mac-1 inside-out activation have remained poorly understood. Phosphorylation of integrin cytoplasmic domains is emerging as an important mechanism of regulating integrin functions. Here we have studied phosphorylation of human CD11b, which takes place on the cytoplasmic Ser1126 in neutrophils. We show that mutation of the serine phosphorylation site leads to inability of Mac-1 to become activated to bind the cellular ligands ICAM-1 and ICAM-2. However, CD11b-mutant cells are fully capable of binding other studied CD11b-ligands, i.e. iC3b and denatured BSA. Activation epitopes expressed in the extracellular domain of the integrin and affinity for soluble ICAM ligands were decreased for the mutated integrin. Additionally, the mutation resulted in inhibition of chemokine-induced migration in a transendothelial assay