title: Interaction of antibodies to proteinase 3 (classic anti-neutrophil cytoplasmic antibody) with human renal tubular epithelial cells: impact on signaling events and inflammatory mediator generation.
abstract: Among the anti-neutrophil cytoplasmic Abs (ANCA), those targeting proteinase 3 (PR3) have a high sensitivity and specificity for Wegener's granulomatosis (WG). A pathogenetic role for these autoantibodies has been proposed due to their capacity of activating neutrophils in vitro. Recently, PR3 was also detected in human renal tubular epithelial cells (TEC). In the present study, the effect of murine monoclonal anti-PR3 Abs (anti-PR3) and purified c-ANCA targeting PR3 from WG serum on isolated human renal tubular cell signaling and inflammatory mediator release was characterized. Priming of TEC with TNF-alpha resulted in surface expression of PR3, as quantified in immunofluorescence studies and by flow cytometry. Moreover, PR3 was immunoprecipitated on surface-labeled TEC. Primed TEC responded to anti-PR3 with a dose- and time-dependent activation of phosphoinositide hydrolysis, resulting in a remarkable accumulation of inositolphosphates. Control IgG was entirely ineffective, whereas PR3-ANCA reproduced the phosphoinositide response. The signaling response was accompanied by a pronounced release of superoxidanion into the cell supernatant. Moreover, large amounts of PGE(2) and, to a lesser extent, of thromboxane B(2), the stable metabolite of TxA(2), were secreted from anti-PR3-stimulated TEC. In parallel, a rise in intracellular cAMP levels was observed, which was blocked by the cyclooxygenase inhibitor indomethacin. We conclude that anti-PR3 Abs directly target renal TECs, thereby provoking pronounced activation of the phosphoinositide-related signal transduction pathway. Associated metabolic events such as the release of reactive oxygen species and lipid mediators may directly contribute to the development of renal lesions and loss of kidney function in WG.
authors.affiliation: Department of Internal Medicine, Institute for Clinical Immunology and Transfusion Medicine, Justus-Liebig-University, Giessen, Germany.
authors.forename: Katja
authors.initials: K
authors.lastname: Hattar
authors.forename: Ulrich
authors.initials: U
authors.lastname: Grandel
authors.forename: Annette
authors.initials: A
authors.lastname: Bickenbach
authors.forename: Andreas
authors.initials: A
authors.lastname: Schwarting
authors.forename: Werner-Johannes
authors.initials: WJ
authors.lastname: Mayet
authors.forename: Jürgen
authors.initials: J
authors.lastname: Bux
authors.forename: Sönke
authors.initials: S
authors.lastname: Jessen
authors.forename: Claus
authors.initials: C
authors.lastname: Fischer
authors.forename: Werner
authors.initials: W
authors.lastname: Seeger
authors.forename: Friedrich
authors.initials: F
authors.lastname: Grimminger
authors.forename: Ulf
authors.initials: U
authors.lastname: Sibelius
mesh_terms: D019268:Antibodies, Antineutrophil Cytoplasmic / Q000378:metabolism* / Q000494:pharmacology; D000911:Antibodies, Monoclonal / Q000378:metabolism / Q000494:pharmacology; D000937:Antigen-Antibody Reactions; D016718:Arachidonic Acid / Q000378:metabolism; D001324:Autoantigens / Q000096:biosynthesis / Q000276:immunology; D002462:Cell Membrane / Q000201:enzymology / Q000276:immunology; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism; D015232:Dinoprostone / Q000378:metabolism; D004847:Epithelial Cells / Q000201:enzymology / Q000276:immunology* / Q000378:metabolism*; D006801:Humans; D006868:Hydrolysis; D007106:Immune Sera / Q000378:metabolism / Q000494:pharmacology; D018836:Inflammation Mediators / Q000378:metabolism*; D007424:Intracellular Fluid / Q000378:metabolism; D007684:Kidney Tubules / Q000201:enzymology / Q000276:immunology* / Q000378:metabolism*; D053146:Myeloblastin; D010716:Phosphatidylinositols / Q000378:metabolism; D012697:Serine Endopeptidases / Q000096:biosynthesis / Q000276:immunology*; D015398:Signal Transduction / Q000276:immunology*; D013481:Superoxides / Q000378:metabolism; D013928:Thromboxane A2 / Q000378:metabolism
chemical_list: D019268:Antibodies, Antineutrophil Cytoplasmic; D000911:Antibodies, Monoclonal; D001324:Autoantigens; D007106:Immune Sera; D018836:Inflammation Mediators; D010716:Phosphatidylinositols; D013481:Superoxides; D016718:Arachidonic Acid; D013928:Thromboxane A2; D000242:Cyclic AMP; D012697:Serine Endopeptidases; D053146:Myeloblastin; D015232:Dinoprostone