title: Coexpression of CD177 and membrane proteinase 3 on neutrophils in antineutrophil cytoplasmic autoantibody-associated systemic vasculitis: anti-proteinase 3-mediated neutrophil activation is independent of the role of CD177-expressing neutrophils.
abstract: OBJECTIVE
  Wegener's granulomatosis (WG) is strongly associated with antineutrophil cytoplasmic autoantibodies (ANCAs) directed against proteinase 3 (PR3). Recent studies have shown that membrane-bound PR3 (mPR3) is differentially expressed and colocalizes with CD177/NB1 on circulating neutrophils. We undertook this study to assess the differential expression of CD177 on neutrophils from patients with ANCA-associated systemic vasculitis (ASV) in comparison with patients with systemic lupus erythematosus (SLE), patients with rheumatoid arthritis (RA), and healthy individuals, and to investigate whether colocalization of mPR3 and CD177 affects anti-PR3-mediated neutrophil activation.
  
  
  METHODS
  Expression of CD177 and mPR3 was analyzed by flow cytometry on isolated neutrophils from patients with ASV (n=53), those with SLE (n=30), those with RA (n=26), and healthy controls (n=31). Neutrophil activation mediated by anti-PR3 antibodies was assessed by measuring the oxidative burst with a dihydrorhodamine assay.
  
  
  RESULTS
  Percentages of CD177-expressing neutrophils were significantly higher in patients with ASV and those with SLE than in healthy controls. In 3 healthy donors, CD177 expression was not detected. After priming with tumor necrosis factor alpha, neutrophils remained negative for CD177 while mPR3 expression was induced. Neutrophils from CD177-negative donors or CD177- neutrophils sorted from donors with bimodal expression were susceptible to anti-PR3-mediated oxidative burst. Variation in the extent of anti-PR3-mediated neutrophil activation among different donors occurred independent of the percentage of CD177-expressing neutrophils.
  
  
  CONCLUSIONS
  Membrane expression of CD177 on circulating neutrophils is increased in patients with ASV and in those with SLE, but not in RA patients. However, primed neutrophils from CD177-negative individuals also express mPR3 and are susceptible to anti-PR3-mediated oxidative burst, suggesting that recruitment of CD177-independent mPR3 is involved in anti-PR3-induced neutrophil activation.
authors.affiliation: Department of Rheumatology and Clinical Immunology, University Medical Center Groningen, Groningen, The Netherlands.
authors.forename: N
authors.initials: N
authors.lastname: Hu
authors.forename: J
authors.initials: J
authors.lastname: Westra
authors.forename: M G
authors.initials: MG
authors.lastname: Huitema
authors.forename: M
authors.initials: M
authors.lastname: Bijl
authors.forename: E
authors.initials: E
authors.lastname: Brouwer
authors.forename: C A
authors.initials: CA
authors.lastname: Stegeman
authors.forename: P
authors.initials: P
authors.lastname: Heeringa
authors.forename: P C
authors.initials: PC
authors.lastname: Limburg
authors.forename: C G M
authors.initials: CG
authors.lastname: Kallenberg
mesh_terms: D019268:Antibodies, Antineutrophil Cytoplasmic / Q000032:analysis* / Q000276:immunology; D001172:Arthritis, Rheumatoid / Q000276:immunology; D002462:Cell Membrane / Q000201:enzymology; D005434:Flow Cytometry; D058851:GPI-Linked Proteins; D014890:Granulomatosis with Polyangiitis / Q000276:immunology; D006801:Humans; D007519:Isoantigens / Q000032:analysis*; D008180:Lupus Erythematosus, Systemic / Q000276:immunology; D008562:Membrane Glycoproteins / Q000032:analysis*; D053146:Myeloblastin / Q000032:analysis* / Q000276:immunology / Q000502:physiology; D018375:Neutrophil Activation / Q000502:physiology*; D009504:Neutrophils / Q000737:chemistry* / Q000276:immunology*; D011956:Receptors, Cell Surface / Q000032:analysis*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D014657:Vasculitis / Q000276:immunology*
chemical_list: D019268:Antibodies, Antineutrophil Cytoplasmic; C407242:CD177 protein, human; D058851:GPI-Linked Proteins; D007519:Isoantigens; D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface; D014409:Tumor Necrosis Factor-alpha; D053146:Myeloblastin