title: Systemic injection of products of activated neutrophils and H2O2 in myeloperoxidase-immunized rats leads to necrotizing vasculitis in the lungs and gut.
abstract: The strong association of anti-neutrophil cytoplasmic antibodies with various forms of systemic vasculitis suggests a role for these autoantibodies in the pathophysiology of systemic vasculitis. In the present study, we tested the hypothesis that release of neutrophil lysosomal enzymes in the presence of an anti-myeloperoxidase (anti-MPO) immune response may underlie the development of systemic vasculitis. Brown Norway rats were immunized with MPO in complete Freund's adjuvant or complete Freund's adjuvant alone. Two weeks after immunization, rats bad developed antibodies to human and rat MPO as measured by enzyme-linked immunosorbent assay. Next, rats were intravenously infused with 400 micrograms of a human neutrophil lysosomal extract containing 200 micrograms of MPO followed by 0.5 ml of a 1 mmol/L solution of H2O2 through a cannula inserted into the right jugular vein. Rats were sacrificed at 4 hours, 24 hours, 7 days, or 14 days, and several organs (lungs, heart, liver, spleen, gut, and kidneys) were examined for vasculitic lesions and inflammatory cell infiltrates. Macroscopically, patchy hemorrhagic spots were observed in the lungs and gut of MPO-immunized rats at days 7 and 14 after systemic infection of the neutrophil lysosomal extract and H2O2. Such changes were not observed at earlier time points or in control immunized rats. Histologically, the lungs of MPO-immunized rats sacrificed at days 7 and 14 showed patchy inflammatory cell infiltrates associated with vasculitis, granuloma formation, giant cells, and foci of hemorrhage. At 14 days, early signs of fibrosis were found with deposition of collagen and proliferation of fibroblasts. Furthermore, a prominent leukocytoclastic vasculitis was found in the small intestine of these rats characterized by fibrinoid necrosis and an extensive neutrophilic infiltrate. No inflammatory changes were found in the other organs studied (heart, liver, spleen, and kidneys). Control immunized rats, sacrificed at days 7 and 14 showed only some small foci of inflammatory infiltrates in the lungs whereas no inflammatory changes were found in the gastrointestinal tract. These studies show that release of products from activated neutrophils in the presence of anti-MPO autoantibodies may be relevant to the pathogenesis of anti-MPO-associated vasculitides.
authors.affiliation: Department of Clinical Immunology, University Hospital Groningen, The Netherlands.
authors.forename: P
authors.initials: P
authors.lastname: Heeringa
authors.forename: P
authors.initials: P
authors.lastname: Foucher
authors.forename: P A
authors.initials: PA
authors.lastname: Klok
authors.forename: M G
authors.initials: MG
authors.lastname: Huitema
authors.forename: J W
authors.initials: JW
authors.lastname: Tervaert
authors.forename: J J
authors.initials: JJ
authors.lastname: Weening
authors.forename: C G
authors.initials: CG
authors.lastname: Kallenberg
mesh_terms: D000818:Animals; D006801:Humans; D006861:Hydrogen Peroxide / Q000008:administration & dosage / Q000378:metabolism*; D007262:Infusions, Intravenous; D007421:Intestine, Small / Q000473:pathology*; D019272:Leukocyte Elastase / Q000008:administration & dosage; D008168:Lung / Q000473:pathology*; D008247:Lysosomes / Q000201:enzymology; D053146:Myeloblastin; D009336:Necrosis; D018375:Neutrophil Activation / Q000276:immunology*; D009504:Neutrophils / Q000378:metabolism*; D009195:Peroxidase / Q000008:administration & dosage / Q000276:immunology*; D051381:Rats; D011914:Rats, Inbred BN; D012697:Serine Endopeptidases / Q000008:administration & dosage; D014657:Vasculitis / Q000139:chemically induced* / Q000473:pathology*
chemical_list: D006861:Hydrogen Peroxide; D009195:Peroxidase; D012697:Serine Endopeptidases; D019272:Leukocyte Elastase; D053146:Myeloblastin