title: Anti-neutrophil cytoplasm antibody IgG subclasses in Wegener's granulomatosis: a possible pathogenic role for the IgG4 subclass.
abstract: A characteristic feature of Wegener's granulomatosis is the presence of antineutrophil cytoplasm antibodies (ANCA) to proteinase 3 (PR3). In vitro, ANCA activate neutrophils by co-ligating PR3 and FcgammaRIIa/IIIb receptors. ANCA are predominantly of the IgG isotype, and IgG1, IgG3 and IgG4 subclasses are particularly represented. To address the pathogenic role of individual ANCA-IgG subclass antibodies, patients' sera were screened using indirect immunofluorescence, enzyme-linked immunosorbent assay (ELISA) and subclass PR3-ELISA to identify patients with high titres of PR3-ANCA within the IgG1, IgG3 or IgG4 subclasses. Unfractionated ANCA-IgG and subclass fractions were isolated by affinity chromatography and compared for their capacities to stimulate superoxide production by primed human neutrophils. Donor neutrophils were analysed for constitutive and induced FcgammaRI expression by flow cytometry. The IgG1, IgG3 and IgG4 subclass fractions, isolated from three different ANCA sera, each stimulated superoxide production from neutrophils derived from multiple donors. Subsequently, IgG4 subclass fractions isolated from a further four ANCA positive sera demonstrated varying abilities to stimulate release of superoxide; unrelated to PR3-ANCA titre, neutrophil donor, or neutrophil FcgammaRI expression. The stimulation of superoxide release by IgG1- and IgG3-ANCA subclass fractions is consistent with the proposed mechanism of co-ligation of PR3 antigen and FcgammaRIIa/IIIb receptors. However, the demonstration of similar activity for the IgG4-ANCA subclass fractions isolated from some sera was unexpected. This activity was independent of neutrophil donor and expression of FcgammaRI, suggesting it was capable of activating neutrophils via constitutively expressed FcgammaRIIa/IIIb or co-ligation of other, unidentified, cell surface molecules.
authors.affiliation: MRC Centre for Immune Regulation, The Medical School, University of Birmingham, Birmingham, UK.
authors.forename: M
authors.initials: M
authors.lastname: Holland
authors.forename: P
authors.initials: P
authors.lastname: Hewins
authors.forename: M
authors.initials: M
authors.lastname: Goodall
authors.forename: D
authors.initials: D
authors.lastname: Adu
authors.forename: R
authors.initials: R
authors.lastname: Jefferis
authors.forename: C O S
authors.initials: CO
authors.lastname: Savage
mesh_terms: D000368:Aged; D019268:Antibodies, Antineutrophil Cytoplasmic / Q000276:immunology*; D000915:Antibody Affinity / Q000276:immunology; D001324:Autoantigens / Q000276:immunology; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect / Q000379:methods; D014890:Granulomatosis with Polyangiitis / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology*; D007371:Interferon-gamma / Q000276:immunology; D008297:Male; D008875:Middle Aged; D053146:Myeloblastin; D018375:Neutrophil Activation / Q000276:immunology; D009504:Neutrophils / Q000276:immunology; D009195:Peroxidase / Q000276:immunology; D017452:Receptors, IgG / Q000032:analysis / Q000276:immunology; D012697:Serine Endopeptidases / Q000276:immunology
chemical_list: D019268:Antibodies, Antineutrophil Cytoplasmic; D001324:Autoantigens; C489017:FCGR1A protein, human; D007074:Immunoglobulin G; D017452:Receptors, IgG; D007371:Interferon-gamma; D009195:Peroxidase; D012697:Serine Endopeptidases; D053146:Myeloblastin