title: Antibodies to proteinase-3 mediate expression of intercellular adhesion molecule-1 (ICAM-1, CD 54).
abstract: We investigated the role of intercellular adhesion molecule-1 (ICAM-1) in the adhesion of polymorphonuclear neutrophils (PMN) to classic antineutrophil cytoplasmic antibody (C-ANCA)-treated endothelial cells, independently of cytokines. Human umbilical vein endothelial cells (HUVEC) grown to confluence in cytokine-free conditions were stimulated with C-ANCA sera and affinity-purified anti-proteinase 3 antibodies (PR3) from Wegener's granulomatosis (WG) patients. Non-activated PMN were added to treated HUVEC and adhesion was measured. In parallel experiments, treated HUVEC were fixed and ICAM-1 and E-selectin were assayed by cyto-ELISA; in other experiments anti-ELAM-1 and anti-ICAM-1 antibodies were assessed. In this in vitro model, adhesion of non-activated PMN to anti-PR3-stimulated HUVEC was enhanced. Adhesion was greater with anti-PR3 antibodies than with control and normal immunoglobulins, and correlated with the level of anti-PR3 antibodies. Neutralization of anti-PR3 antibodies by neutrophil azurophilic granule proteins abolished adhesion. This adhesion increased at the fourth hour after simulation, peaked at the twelfth hour and then decreased. This phenomenon occurred mainly through endothelial expression of ICAM-1 (the main counter-receptor for integrins, involved in firm PMN adhesion and migration) and E-selectin on HUVEC membranes. Anti-adhesion molecule antibodies inhibited this adhesion. This work supports the hypothesis of a direct effect of C-ANCA in endothelial stimulation, namely, on endothelium-PMN adhesion, and strengthens the major role of ICAM-1, directly involved in firm sticking of PMN to HUVEC, besides E-selectin. C-ANCA upregulate endothelial adhesiveness and thus participate in inflammatory reactions by providing endothelial adhesive structures for neutrophils. This might be one of the first steps leading to clinical expression of the disease. These results provide new insights into the pathogenesis of C-ANCA-related diseases.
authors.affiliation: Unit of Rheumatology, Centre Hospitalo-universitaire Xavier Bichat, Paris, France.
authors.forename: M
authors.initials: M
authors.lastname: De Bandt
authors.forename: O
authors.initials: O
authors.lastname: Meyer
authors.forename: J
authors.initials: J
authors.lastname: Hakim
authors.forename: C
authors.initials: C
authors.lastname: Pasquier
mesh_terms: D000906:Antibodies / Q000502:physiology*; D019268:Antibodies, Antineutrophil Cytoplasmic / Q000032:analysis; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D002448:Cell Adhesion; D002478:Cells, Cultured; D016207:Cytokines / Q000097:blood; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism*; D053146:Myeloblastin; D009504:Neutrophils / Q000502:physiology; D012697:Serine Endopeptidases / Q000276:immunology*
chemical_list: D000906:Antibodies; D019268:Antibodies, Antineutrophil Cytoplasmic; D000911:Antibodies, Monoclonal; D016207:Cytokines; D018799:Intercellular Adhesion Molecule-1; D012697:Serine Endopeptidases; D053146:Myeloblastin