Thromb Haemost 2009; 102(06): 1212-1218
DOI: 10.1160/TH09-01-0038
Blood Coagulation, Fibrinolysis and Cellular Haemostasis
Schattauer GmbH

Leukocyte urokinase plasminogen activator receptor and PSGL1 play a role in endogenous arterial fibrinolysis

Xufang Bai
1   Henderson Research Centre and McMaster University, Hamilton, Ontario, Canada
,
Jeffrey I. Weitz
1   Henderson Research Centre and McMaster University, Hamilton, Ontario, Canada
,
Peter L. Gross
1   Henderson Research Centre and McMaster University, Hamilton, Ontario, Canada
› Author Affiliations
Financial support: This work was supported by the Canadian Institutes of Health Research (MOP-68908) with financial support for equipment from Hamilton Health Sciences. PLG is a recipient of the William E. Noonan Research Career Award from Hamilton Health Sciences. JIW holds a Canada Research Chair (Tier I) in Thrombosis and the HSFO/J.F. Mustard Chair in Cardiovascular Research at McMaster University.
Further Information

Publication History

Received: 16 January 2009

Accepted after major revision: 29 September 2009

Publication Date:
28 November 2017 (online)

Summary

Fibrin is an integral component of arterial thrombi. Using a mouse model of arteriolar thrombosis, high-speed fluorescence microscopy reveals that, within minutes, the fibrin content of thrombi rapidly increases and then decreases.The decrease in fibrin coincides with leukocyte binding to the thrombi, a process mediated by the interaction of leukocyte P-selectin glycoprotein ligand-1 (PSGL-1) with P-selectin on the surface of activated platelets. Because leukocytes possess urokinase-type plasminogen activator (uPA) activity,we used mice deficient in uPA or the uPA receptor (uPAR) to explore the contribution of leukocyte associated uPA to the loss of fibrin from these thrombi. Fibrin loss in both uPA-deficient mice and uPAR-deficient mice was reduced compared with that in wild-type controls.Transfusion of leukocytes from wild-type mice into uPAR-deficient mice restored fibrin loss to levels similar to that in wild-type mice. In contrast, transfusion of leukocytes from mice deficient in uPAR or PSGL-1 did not enhance fibrin loss. Thus, fibrin loss from microarteriolar thrombi is mediated, at least in part, by leukocyte-associated uPA in a process that requires leukocyte uPAR and PSGL-1.

 
  • References

  • 1 Ruggeri ZM. Platelets in atherothrombosis. Nat Med 2002; 08: 1227-1234.
  • 2 Moir E, Booth NA, Bennett B. et al. Polymorphonuclear leucocytes mediate endogenous thrombus lysis via a uPA-dependent mechanism. Br J Haematol 2001; 113: 72-80.
  • 3 Moir E, Robbie LA, Bennett B. et al. Polymorphonuclear leucocytes have two opposing roles in fibrinolysis. Thromb Haemost 2002; 87: 1006-1010.
  • 4 Mutch NJ, Moir E, Robbie LA. et al. Localization and identification of thrombin and plasminogen activator activities in model human thrombi by in situ zymography. Thromb Haemost 2002; 88: 996-1002.
  • 5 Gross PL, Furie BC, Merrill-Skoloff G. et al. Leukocyte-versus microparticle-mediated tissue factor transfer during arteriolar thrombus development. J Leukoc Biol 2005; 78: 1318-1326.
  • 6 Celi A, Lorenzet R, Furie B. et al. Platelet-leukocyte-endothelial cell interaction on the blood vessel wall. Semin Hematol 1997; 34: 327-335.
  • 7 Celi A, Pellegrini G, Lorenzet R. et al. P-selectin induces the expression of tissue factor on monocytes. Proc Natl Acad Sci USA 1994; 91: 8767-8771.
  • 8 Carmeliet P, Schoonjans L, Kieckens L. et al. Physiological consequences of loss of plasminogen activator gene function in mice. Nature 1994; 368: 419-424.
  • 9 Bugge TH, Suh TT, Flick MJ. et al. The receptor for urokinase-type plasminogen activator is not essential for mouse development or fertility. J Biol Chem 1995; 270: 16886-16894.
  • 10 Yang J, Hirata T, Croce K. et al. Targeted gene disruption demonstrates that P-selectin glycoprotein ligand 1 (PSGL-1) is required for P-selectin-mediated but not E-selectin-mediated neutrophil rolling and migration. J Exp Med 1999; 190: 1769-1782.
  • 11 Falati S, Gross P, Merrill-Skoloff G. et al. Real-time in vivo imaging of platelets, tissue factor and fibrin during arterial thrombus formation in the mouse. Nat Med 2002; 08: 1175-1181.
  • 12 Celi A, Merrill-Skoloff G, Gross P. et al. Thrombus formation: direct real-time observation and digital analysis of thrombus assembly in a living mouse by confocal and widefield intravital microscopy. J Thromb Haemost 2003; 01: 60-68.
  • 13 Cotter MJ, Norman KE, Hellewell PG. et al. A novel method for isolation of neutrophils from murine blood using negative immunomagnetic separation. Am J Pathol 2001; 159: 473-481.
  • 14 Falati S, Patil S, Gross PL. et al. Platelet PECAM-1 inhibits thrombus formation in vivo. Blood 2006; 107: 535-541.
  • 15 Falati S, Liu Q, Gross P. et al. Accumulation of tissue factor into developing thrombi in vivo is dependent upon microparticle P-selectin glycoprotein ligand 1 and platelet P-selectin. J Exp Med 2003; 197: 1585-1598.
  • 16 Bouchard BA, Tracy PB. The participation of leukocytes in coagulant reactions. J Thromb Haemost 2003; 01: 464-469.
  • 17 Colucci M, Stramaglia AM, Mascolo E. et al. Monocytes, but not endothelial cells, downregulate the anticoagulant activity of activated protein C. Br J Haematol 2001; 112: 519-526.
  • 18 Plescia J, Altieri DC. Activation of Mac-1 (CD11b/CD18)-bound factor X by released cathepsin G defines an alternative pathway of leucocyte initiation of coagulation. Biochem J 1996; 319: 873-879.
  • 19 Sturn DH, Kaneider NC, Feistritzer C. et al. Expression and function of the endothelial protein C receptor in human neutrophils. Blood 2003; 102: 1499-1505.
  • 20 Moroz LA, Gilmore NJ. Fibrinolysis in normal plasma and blood: evidence for significant mechanisms independent of the plasminogen-plasmin system. Blood 1976; 48: 531-545.
  • 21 Plow EF. The major fibrinolytic proteases of human leukocytes. Biochim Biophys Acta 1980; 630: 47-56.
  • 22 Singh I, Burnand KG, Collins M. et al. Failure of thrombus to resolve in urokinase-type plasminogen activator gene-knockout mice: rescue by normal bone marrow-derived cells. Circulation 2003; 107: 869-875.
  • 23 Bdeir K, Murciano JC, Tomaszewski J. et al. Urokinase mediates fibrinolysis in the pulmonary microvasculature. Blood 2000; 96: 1820-1826.
  • 24 Giles AR, Nesheim ME, Herring SW. et al. The fibrinolytic potential of the normal primate following the generation of thrombin in vivo. Thromb Haemost 1990; 63: 476-481.
  • 25 Ploplis VA, Carmeliet P, Vazirzadeh S. et al. Effects of disruption of the plasminogen gene on thrombosis, growth, and health in mice. Circulation 1995; 92: 2585-2593.
  • 26 Schafer K, Konstantinides S, Riedel C. et al. Different mechanisms of increased luminal stenosis after arterial injury in mice deficient for urokinase- or tissue-type plasminogen activator. Circulation 2002; 106: 1847-1852.
  • 27 Kufrin D, Eslin DE, Bdeir K. et al. Antithrombotic thrombocytes: ectopic expression of urokinase-type plasminogen activator in platelets. Blood 2003; 102: 926-933.
  • 28 Ragno P. The urokinase receptor: a ligand or a receptor? Story of a sociable molecule. Cell Mol Life Sci 2006; 63: 1028-1037.
  • 29 Reheman A, Gross P, Yang H. et al. Vitronectin stabilizes thrombi and vessel occlusion but plays a dual role in platelet aggregation. J Thromb Haemost 2005; 03: 875-883.
  • 30 Subramaniam M, Frenette PS, Saffaripour S. et al. Defects in hemostasis in P-selectin-deficient mice. Blood 1996; 87: 1238-1242.
  • 31 Grunewald M, Siegemund A, Grunewald A. et al. Plasmatic coagulation and fibrinolytic system alterations in PNH: relation to clone size. Blood Coagul Fibrinolysis 2003; 14: 685-695.
  • 32 Sloand EM, Pfannes L, Scheinberg P. et al. Increased soluble urokinase plasminogen activator receptor (suPAR) is associated with thrombosis and inhibition of plasmin generation in paroxysmal nocturnal hemoglobinuria (PNH) patients. Exp Hematol 2008; 36: 1616-1624.
  • 33 Pillay V, Dass CR, Choong PF. The urokinase plasminogen activator receptor as a gene therapy target for cancer. Trends Biotechnol 2007; 25: 33-39.
  • 34 Lund IK, Jogi A, Rono B. et al. Antibody-mediated targeting of the urokinase-type plasminogen activator proteolytic function neutralizes fibrinolysis in vivo. J Biol Chem 2008; 283: 32506-32515.
  • 35 Wan H, Liu Z, Xia X. et al. A recombinant antibody-targeted plasminogen activator with high affinity for activated platelets increases thrombolytic potency in vitro and in vivo. Thromb Res 2000; 97: 133-141.
  • 36 Dong N, Da Cunha V, Citkowicz A. et al. P-selectintargeting of the fibrin selective thrombolytic Desmodus rotundus salivary plasminogen activator alpha1. Thromb Haemost 2004; 92: 956-965.
  • 37 Ramachandran V, Williams M, Yago T. et al. Dynamic alterations of membrane tethers stabilize leukocyte rolling on P-selectin. Proc Natl Acad Sci USA 2004; 101: 13519-13524.