Antineutrophil cytoplasmic antibodies induce human monocytes to produce oxygen radicals in vitro.
OBJECTIVE
Antineutrophil cytoplasmic antibodies (ANCA) are believed to play a pathogenetic role in necrotizing small-vessel vasculitis. While the involvement of neutrophils in this disease has been extensively studied in vitro, we undertook to analyze thoroughly the contribution of monocytes to tissue destruction in systemic vasculitis.


METHODS
Monocytes obtained from normal human individuals were stimulated by ANCA isolated from patients with active vasculitis. The formation of oxygen radicals was measured by a fluorometric assay using 2',7'-dichlorofluorescin diacetate.


RESULTS
ANCA induced monocytes to produce oxygen radicals, resulting in a mean 43% increase (range 21-84%) in oxygen radical formation compared with normal IgG. The formation of reactive oxygen species was time and concentration dependent and was also induced by ANCA F(ab')2 fragments. Normal nonspecific IgG or their corresponding F(ab')2 fragments induced no release or very little release of oxygen radicals. Preincubation of monocytes with the Fcy receptor type II-blocking monoclonal antibody IV.3 before addition of ANCA greatly reduced formation of oxygen radicals. Using ligand affinity chromatography with proteinase 3 (PR3) and myeloperoxidase (MPO), ANCA were further purified by depletion of patient IgG. The stimulation of monocytes with these pure PR3- and MPO-ANCA confirmed that cellular activation was specifically induced by ANCA.


CONCLUSIONS
These results show that ANCA induce the formation of reactive oxygen species in human monocytes. These findings support the notion that ANCA specifically activate monocytes by several mechanisms to participate in the inflammatory process of ANCA-associated vasculitis.
Weidner
S
S
University of Erlangen-Nürnberg, Germany.
Neupert
W
W
Goppelt-Struebe
M
M
Rupprecht
H D
HD
D019268:Antibodies, Antineutrophil Cytoplasmic / Q000494:pharmacology*; D000918:Antibody Specificity; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000494:pharmacology; D066298:In Vitro Techniques; D017493:Leukocyte Common Antigens / Q000032:analysis; D018950:Lipopolysaccharide Receptors / Q000032:analysis; D009000:Monocytes / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D017452:Receptors, IgG / Q000378:metabolism; D014657:Vasculitis / Q000276:immunology* / Q000378:metabolism*
D019268:Antibodies, Antineutrophil Cytoplasmic; D007140:Immunoglobulin Fab Fragments; D018950:Lipopolysaccharide Receptors; D017382:Reactive Oxygen Species; D017452:Receptors, IgG; D017493:Leukocyte Common Antigens