title: Priming by tumor necrosis factor-alpha of human neutrophil NADPH-oxidase activity induced by anti-proteinase-3 or anti-myeloperoxidase antibodies.
abstract: Anti-proteinase-3 (anti-PR3) or anti-myeloperoxidase (anti-MPO) antibodies are capable of activating human neutrophils primed by TNF-alpha in vitro. We described previously the involvement of FcgammaRIIa and beta(2) integrins in this neutrophil activation. In the literature, the requirement of TNF priming has been attributed to an effect of TNF-alpha on the expression of PR3 or MPO on the cell surface. Under our experimental conditions, TNF-alpha (2 ng/ml) increased the binding of the antibody against PR3, whereas binding of the antibody against MPO could hardly be detected, not even after TNF-alpha treatment. The aim of this study was to consider (an)other(s) role(s) for TNF-alpha in facilitating the NADPH-oxidase activation by these antibodies. We demonstrate the early mobilization of the secretory vesicles as a result of TNF-induced increase in intracellular-free calcium ions, the parallel colocalization of gp91(phox), the main component of the NADPH oxidase with beta(2) integrins and FcgammaRIIa on the neutrophil surface, and the FcgammaRIIa clustering upon TNF priming. TNF-alpha also induced redistribution of FcgammaRIIa to the cytoskeleton in a dose- and time-dependent manner. Moreover, blocking CD18 MHM23 antibody, cytochalasin B, and D609 (an inhibitor of phosphatidylcholine phospholipase C) inhibited this redistribution and the respiratory burst in TNF-treated neutrophils exposed to anti-PR3 or anti-MPO antibodies. Our results indicate direct effects of TNF-alpha in facilitating neutrophil activation by these antibodies and further support the importance of cytoskeletal rearrangements in this priming process.
authors.affiliation: Faculté des Sciences Pharmaceutiques et Biologiques, Université de Lille-2, 3 rue du Professeur Laguesse, 59006 Lille cedex, France. dominique.reumaux@libertysurf.fr
authors.forename: Dominique
authors.initials: D
authors.lastname: Reumaux
authors.forename: Peter L
authors.initials: PL
authors.lastname: Hordijk
authors.forename: Patrick
authors.initials: P
authors.lastname: Duthilleul
authors.forename: Dirk
authors.initials: D
authors.lastname: Roos
mesh_terms: D000906:Antibodies / Q000276:immunology / Q000494:pharmacology*; D015703:Antigens, CD / Q000276:immunology / Q000378:metabolism; D001952:Bridged-Ring Compounds / Q000494:pharmacology; D018821:CD18 Antigens / Q000276:immunology / Q000378:metabolism; D002478:Cells, Cultured; D003572:Cytochalasins / Q000494:pharmacology; D003599:Cytoskeleton / Q000276:immunology / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects* / Q000276:immunology; D006801:Humans; D008562:Membrane Glycoproteins / Q000096:biosynthesis* / Q000276:immunology; D053146:Myeloblastin / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000276:immunology; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases / Q000096:biosynthesis* / Q000276:immunology; D018375:Neutrophil Activation / Q000187:drug effects* / Q000276:immunology; D009504:Neutrophils / Q000201:enzymology* / Q000276:immunology; D009636:Norbornanes; D009195:Peroxidase / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000276:immunology; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D017452:Receptors, IgG / Q000276:immunology / Q000378:metabolism; D013859:Thiocarbamates; D013871:Thiones / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology / Q000494:pharmacology*; D010738:Type C Phospholipases / Q000037:antagonists & inhibitors / Q000276:immunology
chemical_list: D000906:Antibodies; D015703:Antigens, CD; D001952:Bridged-Ring Compounds; D018821:CD18 Antigens; D003572:Cytochalasins; C080568:Fc gamma receptor IIA; D008562:Membrane Glycoproteins; D009636:Norbornanes; D010726:Phosphodiesterase Inhibitors; D017452:Receptors, IgG; D013859:Thiocarbamates; D013871:Thiones; D014409:Tumor Necrosis Factor-alpha; C031218:cytochalasin A; C046498:tricyclodecane-9-yl-xanthogenate; D009195:Peroxidase; C056558:CYBB protein, human; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases; D010738:Type C Phospholipases; C105598:phosphatidylcholine-specific phospholipase C; D053146:Myeloblastin