title: Antineutrophil cytoplasm autoantibodies from patients with systemic vasculitis activate neutrophils through distinct signaling cascades: comparison with conventional Fcgamma receptor ligation.
abstract: In systemic vasculitis, interactions between antineutrophil cytoplasm autoantibodies (ANCAs) and neutrophils initiate endothelial and vascular injury. ANCAs directed against either myeloperoxidase (MPO) or proteinase 3 (PR3) can activate cytokine-primed neutrophils by binding cell surface-expressed MPO or PR3, with the concurrent engagement of Fcgamma receptors (FcgammaR). Because roles for phospholipase D (PLD) and phosphatidylinositol 3 kinase (PI3K) have been demonstrated in FcgammaR activation of neutrophils, this study investigated the hypothesis that ANCA stimulation of neutrophils involved a similar engagement of FcgammaR and activation of PLD and PI3K. Pretreatment of tumor necrosis factor (TNF) alpha-primed neutrophils with antibodies against FcgammaRII and FcgammaRIII inhibited MPO-ANCA and PR3-ANCA induced superoxide generation, confirming that FcgammaR ligation is involved in ANCA-mediated neutrophil activation. However, although stimulation of TNF-alpha-primed neutrophils by conventional FcgammaR ligation, either using antibody-mediated cross-linking of FcgammaR or aggregated IgG, induced PLD activation, ANCA stimulation did not. Moreover, although ANCA-induced neutrophil activation results in significant PI3K activation-as assessed by phosphatidylinositol 3,4,5-triphosphate generation-conventional FcgammaR ligation, but not ANCA, activates the p85/p110 PI3K subtype. Inhibition of ANCA-induced superoxide generation with pertussis toxin suggests that ANCAs activate the p101/p110gamma PI3K isoform. In addition, the kinetics of activation of protein kinase B differs between conventional FcgammaR ligation and ANCA stimulation of neutrophils. These results demonstrate that though ligation of FcgammaRIIa and FcgammaRIIIb may be necessary, it is likely that ANCAs require other membrane cofactors for neutrophil activation.
authors.affiliation: Renal Immunobiology, MRC Centre for Immune Regulation, the Department of Biochemistry, and the Institute for Cancer Studies, The Medical School, University of Birmingham, United Kingdom.
authors.forename: A
authors.initials: A
authors.lastname: Ben-Smith
authors.forename: S K
authors.initials: SK
authors.lastname: Dove
authors.forename: A
authors.initials: A
authors.lastname: Martin
authors.forename: M J
authors.initials: MJ
authors.lastname: Wakelam
authors.forename: C O
authors.initials: CO
authors.lastname: Savage
mesh_terms: D000730:Androstadienes / Q000494:pharmacology; D019268:Antibodies, Antineutrophil Cytoplasmic / Q000276:immunology*; D001324:Autoantigens / Q000276:immunology; D001327:Autoimmune Diseases / Q000276:immunology*; D002867:Chromones / Q000494:pharmacology; D004075:Diglycerides / Q000096:biosynthesis; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D007527:Isoenzymes / Q000502:physiology; D008970:Molecular Weight; D009025:Morpholines / Q000494:pharmacology; D053146:Myeloblastin; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000187:drug effects / Q000276:immunology*; D009195:Peroxidase / Q000276:immunology; D037342:Pertussis Toxin; D019869:Phosphatidylinositol 3-Kinases / Q000502:physiology*; D018129:Phosphatidylinositol Phosphates / Q000096:biosynthesis; D010739:Phospholipase D / Q000502:physiology*; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt; D017452:Receptors, IgG / Q000276:immunology*; D016897:Respiratory Burst / Q000187:drug effects; D012697:Serine Endopeptidases / Q000276:immunology; D015398:Signal Transduction / Q000187:drug effects; D013481:Superoxides / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D014657:Vasculitis / Q000276:immunology*; D010566:Virulence Factors, Bordetella / Q000494:pharmacology; D000077191:Wortmannin
chemical_list: D000730:Androstadienes; D019268:Antibodies, Antineutrophil Cytoplasmic; D001324:Autoantigens; D002867:Chromones; D004075:Diglycerides; D004791:Enzyme Inhibitors; D007074:Immunoglobulin G; D007527:Isoenzymes; D009025:Morpholines; D053146:Myeloblastin; D009240:N-Formylmethionine Leucyl-Phenylalanine; D009195:Peroxidase; D037342:Pertussis Toxin; D019869:Phosphatidylinositol 3-Kinases; D018129:Phosphatidylinositol Phosphates; D010739:Phospholipase D; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins; D051057:Proto-Oncogene Proteins c-akt; D017452:Receptors, IgG; D012697:Serine Endopeptidases; D013481:Superoxides; D014409:Tumor Necrosis Factor-alpha; D010566:Virulence Factors, Bordetella; D000077191:Wortmannin; C060974:phosphatidylinositol 3,4,5-triphosphate; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one