title: Activated neutrophils express proteinase 3 on their plasma membrane in vitro and in vivo.
abstract: Apart from the diagnostic value of anti-neutrophil cytoplasmic antibodies (ANCA), their detailed characterization and that of their corresponding antigens have opened new ways for the exploration of the pathogenesis of primary systemic vasculitis. ANCA are now thought to play an important functional role via activation of phagocytic cells (e.g. polymorphonuclear neutrophils (PMN)). In this study we examined the mechanisms by which ANCA could gain access to proteinase 3 (PR3) in intact PMN, at two levels: ex vivo by analysing the presence of PR3 on the plasma membrane of PMN from patients with ANCA-associated vasculitis, and in vitro by stimulation of PMN using different cytokines, including recombinant tumour necrosis factor-alpha (rhTNF-alpha) and two forms of IL-8 (produced by monocytic and endothelial cells). Using immunocytochemical staining techniques (FACS and immunoelectronmicroscopy) PR3 has been detected on the plasma membrane of PMN from patients with active ANCA-associated vasculitis. However, this phenomenon is also seen in patients with sepsis who do not have ANCA. In addition, TNF-alpha and both forms of IL-8 act synergistically and induce a translocation of PR3 from the intragranular loci to the cell surface of PMN. These results provide strong evidence for the hypothesis that ANCA are directly pathogenic by binding to PR3 which is expressed on the cell surface of primed/activated PMN.
authors.affiliation: Rhematological Department, Medical University Lübeck, Germany.
authors.forename: E
authors.initials: E
authors.lastname: Csernok
authors.forename: M
authors.initials: M
authors.lastname: Ernst
authors.forename: W
authors.initials: W
authors.lastname: Schmitt
authors.forename: D F
authors.initials: DF
authors.lastname: Bainton
authors.forename: W L
authors.initials: WL
authors.lastname: Gross
mesh_terms: D019268:Antibodies, Antineutrophil Cytoplasmic; D015703:Antigens, CD / Q000032:analysis; D001323:Autoantibodies / Q000032:analysis; D002462:Cell Membrane / Q000201:enzymology; D014890:Granulomatosis with Polyangiitis / Q000097:blood*; D006801:Humans; D016209:Interleukin-8 / Q000494:pharmacology; D053146:Myeloblastin; D009504:Neutrophils / Q000187:drug effects / Q000201:enzymology*; D010980:Platelet Membrane Glycoproteins / Q000032:analysis; D012697:Serine Endopeptidases / Q000032:analysis*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D060149:Tetraspanin 30; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D019268:Antibodies, Antineutrophil Cytoplasmic; D015703:Antigens, CD; D001323:Autoantibodies; C554586:CD63 protein, human; D016209:Interleukin-8; D010980:Platelet Membrane Glycoproteins; D060149:Tetraspanin 30; D014409:Tumor Necrosis Factor-alpha; D012697:Serine Endopeptidases; D053146:Myeloblastin; D013755:Tetradecanoylphorbol Acetate