COMPLEMENT RECEPTOR-3 (CR3, MAC-1, INTEGRIN-ALPHA(M)BETA(2), CD11B/CD18) IS REQUIRED FOR TYROSINE PHOSPHORYLATION OF PAXILLIN IN ADHERENT AND NONADHERENT NEUTROPHILS

Expression of the leukocyte (beta(2)) integrins is required for many functions of activated neutrophils (PMN), even when there is no recognized Ligand for any beta(2) integrin. To investigate the hypothesis that beta(2) integrins may be involved in a signal transduction pathway related to cytoskeletal reorganization, we examined whether beta(2) integrins have a role in tyrosine phosphorylation of the cytoskeletal protein paxillin. Treatment of PMN in suspension with phorbol esters, f-Met-Leu-Phe, and TNF-alpha resulted in paxillin tyrosine phosphorylation. However, treatment of beta(2)-deficient (LAD) PMN failed to induce paxillin tyrosine phosphorylation. Normal PMN phosphorylated paxillin in response to adhesion to immune complexes, while the LAD PMN did not. Adhesion of phorbol ester activated-LAD PMN to the extracellular matrix proteins fibronectin, laminin, and vitronectin faded to induce paxillin tyrosine phosphorylation. Treatment of activated normal PMN with mAb directed against the beta(2) integrin alpha chains demonstrated that CR3 (alpha(M) beta(2)) was required for paxillin phospholylation. Transfection of the cell Line K562 with CR3 confirmed that CR3 ligation resulted in paxillin tyrosine phosphorylation. As a control, K562 transfected with CR2 (CD21) which bound equally avidly to the same complement C3-derived ligand (C3bi) as the CR3 transfectants, showed no enhanced tyrosine phosphorylation of paxillin upon receptor ligation. While both CR2 and CR3 transfectants showed efficient adhesion to a C3bi-coated surface, only the CR3 transfectants spread during adhesion and phosphorylated paxillin. Together these data demonstrate that CR3 is required for paxillin phosphorylation during activation of both adherent and nonadherent PMN. Even PMN activated in suspension or by adhesion to immune complexes, when no CR3 ligand is apparent, still require CR3 for a signal transduction pathway leading to paxillin tyrosine phosphorylation. This pathway is likely to be important for PMN function in inflammation and host defense.