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Blood, 1 October 2007, Vol. 110, No. 7, pp. 2432-2439.
Prepublished online as a Blood First Edition Paper on July 2, 2007; DOI 10.1182/blood-2007-02-069997.


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HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY

Activation of plasminogen into plasmin at the surface of endothelial microparticles: a mechanism that modulates angiogenic properties of endothelial progenitor cells in vitro

Romaric Lacroix1, Florence Sabatier1, Agnès Mialhe1, Agnès Basire1, Ralph Pannell2, Hélène Borghi3, Stephane Robert1, Edouard Lamy1, Laurent Plawinski4, Laurence Camoin-Jau1, Victor Gurewich2, Eduardo Angles-Cano4, and Françoise Dignat-George1

1 Unité Mixte de Recherche S 608 (UMR-S 608), Institut National de la Santé et de la Recherche Médicale (INSERM), Université de la Méditerranée, Unité de Formation et de Recherche (UFR) de Pharmacie, Marseille, France; 2 Vascular Research Laboratory, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA; 3 Service de Microscopie Electronique, UFR de Médecine, Université de la Méditerranée, Marseille, France; 4 INSERM, Cyceron, Université de Caen, Caen, France

The regulation of plasmin generation on cell surfaces is of critical importance in the control of vascular homeostasis. Cell-derived microparticles participate in the dissemination of biological activities. However, their capacity to promote plasmin generation has not been documented. In this study, we show that endothelial microparticles (EMPs) from tumor necrosis factor {alpha} (TNF{alpha})–stimulated endothelial cells served as a surface for the generation of plasmin. The generation of plasmin involved expression of urokinase-type plasminogen activator (uPA) and its receptor (uPAR) at the surface of EMPs and was further increased by their ability to bind exogenous uPA on uPAR. Plasminogen was activated at the surface of EMPs in a dose-dependent, saturable, and specific manner as indicated by the inhibition of plasmin formation by {epsilon}-amino-caproic acid ({epsilon}-ACA) and carboxypeptidase B. EMP-induced plasmin generation affects tube formation mediated by endothelial progenitor cells. However, low amounts of EMPs increased tube formation, whereas higher concentrations inhibited it. Prevention of these effects by inhibitors of either uPA or plasmin underscore the key role of EMP-induced plasmin generation. In conclusion, we demonstrated that EMPs act as vectors supporting efficient plasmin generation and dissemination, a new pathway in the regulation of endothelial proteolytic activities with potential involvement in inflammation, angiogenesis, and atherosclerosis.


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