title: Integrins and cytokines activate nuclear transcription factor-kappaB in human neutrophils.
abstract: Neutrophil adhesion to extracellular matrix is necessary for an effective inflammatory response. Adhesion may accelerate neutrophil activation by affecting intracellular signaling pathways. The nuclear transcription factor kappaB (NF-kappaB) controls several cellular functions, including inflammation, proliferation, and cell survival. We explored the role of adhesion in NF-kappaB activation in human neutrophils. Cells were stimulated with tumor necrosis factor-alpha (TNF-alpha), granulocyte macrophage-colony-stimulating factor (GM-CSF), interleukin-8 (IL-8), and formyl-methionyl-leucyl-phenylalanine (fMLP). All four initiated neutrophil adherence to and spreading on fibronectin. GM-CSF and IL-8 did not activate NF-kappaB in suspended neutrophils but rapidly activated NF-kappaB under adherent conditions on matrix, as shown by IkappaB kinase activity assay, IkappaBalpha degradation, electromobility shift assay, and quantitative reverse transcriptase-PCR. In contrast, TNF-alpha activated NF-kappaB both in suspended cells and adherent cells. fMLP did not activate NF-kappaB in either suspended or adherent cells. Specific beta(2) integrin blockade prevented NF-kappaB activation by GM-CSF and IL-8 on fibronectin. Co-stimulating CD18 and CD11b with activating antibodies resulted in NF-kappaB activation by GM-CSF and IL-8 in suspended cells. We inhibited actin polymerization with cytochalasin and blocked the non-receptor kinase Syk with piceatannol. Both maneuvers prevented the co-stimulatory NF-kappaB-activating signal by beta(2) integrins. Thus, in addition to beta(2) integrin ligand binding, NF-kappaB activation depended on the formation of the receptor-associated intracellular focal adhesion complex. We conclude that beta(2) integrins may provide co-stimulatory signals allowing some soluble mediators to activate the NF-kappaB pathway even when they are not capable of doing so in suspension. This effect may become important when human neutrophils leave the circulating blood and migrate through extracellular matrix during inflammation.
authors: Medical Faculty of the Charité, Department of Nephrology and Hypertension, Franz Volhard Clinic, HELIOS-Klinikum-Berlin and Max Delbrück Center for Molecular Medicine, 13125 Berlin, Germany. kettritz@fvk-berlin.de
authors: Ralph
authors: R
authors: Kettritz
authors: Mira
authors: M
authors: Choi
authors: Susanne
authors: S
authors: Rolle
authors: Maren
authors: M
authors: Wellner
authors: Friedrich C
authors: FC
authors: Luft
mesh_terms: D016207:Cytokines / Q000378:metabolism*; D006801:Humans; D016023:Integrins / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism*; D018375:Neutrophil Activation; D020556:Neutrophil Infiltration; D009504:Neutrophils / Q000378:metabolism*; D015854:Up-Regulation
chemical_list: D016207:Cytokines; D016023:Integrins; D016328:NF-kappa B