Endothelial tissue factor stimulation by proteinase 3 and elastase.
In ANCA-associated vasculitis the activation of primed leucocytes by autoantibodies with subsequent release of proteases such as myeloperoxidase (MPO), proteinase 3 (PR3) and elastase is thought to play an important pathogenetic role. Whether these proteases contribute to the vascular lesions by stimulating the procoagulant activity of these cells is unknown. Tissue factor (TF) expression and activity were investigated in human umbilical vein endothelial cells after stimulation with MPO, PR3 and elastase. TF activity was measured using a one-stage clotting assay. Polyclonal antibodies to TF were used to prove specificity. TF mRNA was detected by reverse transcriptase-polymerase chain reaction. PR3 and elastase led to a significant increase in TF mRNA expression and increased activity. The stimulation was not mediated by IL-1. The stimulatory effect of PR3 did not depend on its proteolytic activity (no inhibition by alpha-1-antitrypsin), whereas the effect of elastase was blocked by alpha-1-antitrypsin. MPO had no effect on TF activity. These results show that PR3 and elastase stimulate TF expression in human endothelial cells. In ANCA-associated vasculitis the increased release of proteases may contribute to the development of microthrombi and consecutive necrosis.
Haubitz
M
M
Department of Nephrology, Medical School Hannover, Germany. Haubitz.Marion@MH-Hannover.de
Gerlach
M
M
Kruse
H J
HJ
Brunkhorst
R
R
D019268:Antibodies, Antineutrophil Cytoplasmic / Q000378:metabolism; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000187:drug effects* / Q000378:metabolism*; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D053590:Interleukin 1 Receptor Antagonist Protein; D007375:Interleukin-1 / Q000378:metabolism; D053146:Myeloblastin; D010196:Pancreatic Elastase / Q000378:metabolism / Q000494:pharmacology*; D009195:Peroxidase / Q000378:metabolism / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012697:Serine Endopeptidases / Q000378:metabolism / Q000494:pharmacology*; D012795:Sialoglycoproteins / Q000494:pharmacology; D013925:Thromboplastin / Q000235:genetics / Q000378:metabolism*; D014657:Vasculitis / Q000209:etiology / Q000276:immunology / Q000378:metabolism
D019268:Antibodies, Antineutrophil Cytoplasmic; C507763:IL1RN protein, human; D053590:Interleukin 1 Receptor Antagonist Protein; D007375:Interleukin-1; D012333:RNA, Messenger; D012795:Sialoglycoproteins; D013925:Thromboplastin; D009195:Peroxidase; D012697:Serine Endopeptidases; D010196:Pancreatic Elastase; D053146:Myeloblastin