Polymorphonuclear Leucocytes Have Two Opposing Roles in Fibrinolysis

E. Moir; L. A. Robbie; B. Bennett; N. A. Booth

doi:10.1055/s-0037-1613125

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2002; 87(06): 1006-1010
DOI: 10.1055/s-0037-1613125

Review Article

Schattauer GmbH

Polymorphonuclear Leucocytes Have Two Opposing Roles in Fibrinolysis

E. Moir

¹Department of Medicine & Therapeutics, University of Aberdeen, Institute of Medical Sciences, Aberdeen, Scotland, UK

²Department of Molecular & Cell Biology, University of Aberdeen, Institute of Medical Sciences, Aberdeen, Scotland, UK

,

L. A. Robbie

¹Department of Medicine & Therapeutics, University of Aberdeen, Institute of Medical Sciences, Aberdeen, Scotland, UK

²Department of Molecular & Cell Biology, University of Aberdeen, Institute of Medical Sciences, Aberdeen, Scotland, UK

,

B. Bennett

¹Department of Medicine & Therapeutics, University of Aberdeen, Institute of Medical Sciences, Aberdeen, Scotland, UK

,

N. A. Booth

²Department of Molecular & Cell Biology, University of Aberdeen, Institute of Medical Sciences, Aberdeen, Scotland, UK

› Author Affiliations

Abstract

Summary

Polymorphonuclear leucocytes (PMN) are important in the resolution of human thrombi, with u-PA as a key player. We have shown that the u-PA activity of PMN depends on the presence of plasma; the study presented here provides an explanation for that requirement. Here we show that PMN degraded scu-PA and also tcu-PA, t-PA and plasmin, resulting in loss of fibrinolytic activity. Plasma protected against this degradation; α₁-antitrypsin was identified as a protective factor. Purified human neutrophil elastase mirrored the effects of PMN, again neutralized by plasma inhibitors. These findings illustrate the dual role of PMN in the breakdown of thrombi, in that they contribute both u-PA, which lyses fibrin, and other proteases, including elastase, which can cleave fibrin and plasminogen activators/plasmin. Similarly, plasma can potentiate fibrinolysis by neutralization of PMN elastase, in addition to direct inhibition of fibrinolytic proteases. Our previous studies show that PMN in thrombi are mostly pro-fibrinolytic; the antifibrinolytic role defined here may be important in other pathologies where fibrin persists.

Keywords

Polymorphonuclear leucocytes - elastase - α₁-antitrypsin - u-PA

Full Text

References

References
1 Granelli-Piperno A, Vassalli JD, Reich E. Secretion of plasminogen activator by human polymorphonuclear leucocytes. Modulation by glucocorticoids and other effectors. J Exp Med 1977; 146: 1693-706.
2 Heiple JM, Ossowski L. Human neutrophil plasminogen activator is localised in specific granules and is translocated to the cell surface by exocytosis. J Exp Med 1986; 164: 826-40.
3 Pedersen TE, Plesner T, Horn T, Hoyer-Hansen G, Sorensen S, Hansen NE. Subcellular distribution of urokinase and urokinase receptor in human neutrophils determined by immunoelectron microscopy. Ultrastruct Pathol 2000; 24: 175-82.
4 Moroz LA, Sniderman AD, Marpole DGF. Leucocyte-protease interactions in fibrinolysis. N Engl J Med 1979; 301: 1100-4.
5 Moir E, Booth NA, Bennett B, Robbie LA. Polymorphonuclear leucocytes mediate spontaneous thrombus lysis via a u-PA-dependent mechanism. Brit J Haem 2001; 113: 72-80.
6 Ohlssen K, Olsson I. The neutral proteases of human granulocytes: Isolation and partial characterisation of granulocyte elastase. Eur J Biochem 1976; 42: 519-27.
7 Rindler-Ludwig R, Braunsteiner H. Cationic proteins from human neutrophil granulocytes: Evidence for their chymotrypsin-like properties. Biochim Biophys Acta 1975; 379: 606-17.
8 Plow EF, Edgington T. An alternative pathway for fibrinolysis. J Clin Invest 1975; 56: 30-8.
9 Plow EF. The major fibrinolytic proteases of human leucocytes. Biochim Biophys Acta 1980; 630: 47-56.
10 Schmitt M, Kanayama N, Henschen A, Hollrieder A, Hafter R, Gulba DC, Janicke F, Graeff H. Elastase released from human granulocytes stimulated with N-formyl-chemotactic peptide prevents activation of tumor cell pro-urokinase (pro-uPA). FEBS Lett 1989; 255: 83-8.
11 Sottrup-Jensen L, Claeys H, Zajdel M, Pedersen TE, Magnusson S. The primary structure of human plasminogen: Isolation of two lysine-binding fragments and one “mini-plasminogen” by elastase-catalysed-specific proteolysis. Prog Chem Fibrin Thrombol 1978; 03: 191-6.
12 Moroz LA. Mini-plasminogen: a mechanism for leukocyte modulation of plasminogen activation by urokinase. Blood 1981; 58: 97-104.
13 Janoff A. Neutrophil proteases in inflammation. Ann Rev Med 1972; 23: 177-89.
14 Baugh R, Travis J. Human leukocyte granule elastase; rapid purification and characterization. Biochem 1976; 15: 836-48.
15 Beatty K, Bieth JG, Travis J. Kinetics of the association of serine proteinases with native and oxidised a ₁-proteinase inhibitor and a ₁-antichymotrypsin. J Biol Chem 1980; 255: 3931-4.
16 Robbie LA, Booth NA, Croll AM, Bennett B. The roles of α₂-antiplasmin and plasminogen activator inhibitor-1 (PAI-1) in the inhibition of clot lysis. Thromb Haemost 1993; 70: 301-6.
17 McWilliam NA, Robbie LA, Booth NA, Bennett B. Plasminogen activator in acute myeloid leukaemic marrow; u-PA in contrast to t-PA in normal marrow. Brit J Haem 1998; 101: 626-31.
18 Booth NA, Anderson JA, Bennett B. Plasminogen activators in alcoholic cirrhosis: demonstration of increased tissue-type and urokinase-type activator. J Clin Path 1984; 37: 772-7.
19 Navia MA, McKeever BM, Springer JP, Lin T-Y, Williams HR, Fluder EM, Dorn CP, Hoogsteen K. Structure of human neutrophil elastase in complex with a peptide chloromethyl ketone inhibitor at 1.84-A resolution. Proc Natl Acad Sci USA 1989; 86: 7-11.
20 Ritchie H, Robbie LA, Kinghorn S, Exley R, Booth NA. Monocyte plasminogen activator inhibitor 2 (PAI-2) inhibits u-PA mediated fibrinolysis and is cross-linked to fibrin. Thromb Haemost 1999; 81: 96-103.
21 Robbie LA, Young SP, Bennett B, Booth NA. Thrombi formed in a Chandler loop mimic human arterial thrombi in structure and PAI-1 content and distribution. Thromb Haemost 1997; 77: 510-5.
22 Kirchhofer D, Riederer MA, Baumgartner HR. Specific accumulation of circulating monocytes and polymorphonuclear leucocytes on platelet thrombi in a vascular injury model. Blood 1997; 89: 1270-8.
23 Hagberg IA, Roald HE, Lyberg T. Adhesion of leucocytes to growing arterial thrombi. Thromb Haemost 1998; 80: 852-8.
24 Machovich R, Owen WG. An elastase-dependent pathway of plasminogen activation. Biochem 1989; 28: 4517-22.
25 Brouwer MS, Harpel PC. Proteolytic cleavage and inactivation of a ₂-plasmin inhibitor and C1-inactivator by human polymorphonuclear leukocyte elastase. J Biol Chem 1982; 257: 9849-54.
26 Levin EG, Santell L. Association of a plasminogen activator inhibitor (PAI-1) with the growth substratum and membrane of human endothelial cells. J Cell Biol 105: 2543-9.