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Thromb Haemost 2017; 117(12): 2300-2311
DOI: 10.1160/TH17-04-0266
Coagulation and Fibrinolysis
Schattauer GmbH Stuttgart

Myeloperoxidase Is a Negative Regulator of Phospholipid-Dependent Coagulation

Lennart Beckmann
,
Christina Dicke
,
Brigitte Spath
,
Carina Lehr
,
Bianca Sievers
,
Anna Klinke
,
Stephan Baldus
,
Volker Rudolph
,
Florian Langer
Further Information

Publication History

19 April 2017

08 September 2017

Publication Date:
06 December 2017 (online)

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Abstract

Myeloperoxidase (MPO) is a cationic heme enzyme stored in neutrophilic polymorphonuclear leukocytes (PMNs) that has recently been implicated in inflammatory cell signaling and tissue damage. Although PMNs play a critical role in both innate immunity and vascular thrombosis, no previous study has systematically investigated the effect of MPO on blood coagulation. Here, we show that PMN-derived MPO inhibits the procoagulant activity (PCA) of lipidated recombinant human tissue factor (rhTF) in a time- and concentration-dependent manner that involves, but is not entirely dependent on the enzyme's catalytic activity. Similarly, MPO together with its substrate, H2O2, inhibited the PCA of plasma microvesicles isolated from lipopolysaccharide (LPS)-stimulated whole blood, an effect additive to that of a function blocking TF antibody. Treatment of whole blood with LPS or phorbol-myristate-acetate dramatically increased MPO plasma levels, and co-incubation with 4-ABAH, a specific MPO inhibitor, significantly enhanced the PCA in plasma supernatants. MPO and MPO/H2O2 also inhibited the PCA of activated platelets and purified phospholipids (PLs), suggesting that modulation of negatively charged PLs, i.e., phosphatidylserine, rather than direct interference with the TF/FVIIa initiation complex was involved. Consistently, pretreatment of activated platelets with MPO or MPO/H2O2 attenuated the subsequent binding of lactadherin, which specifically recognizes procoagulant PS on cell membranes. Finally, endogenously released MPO regulated the PCA of THP1 cells in an autocrine manner dependent on the binding to CD11b/CD18 integrins. Collectively, these findings indicate that MPO is a negative regulator of PL-dependent coagulation and suggest a more complex role of activated PMNs in haemostasis and thrombosis.

Keywords

myeloperoxidase - leukocyte activation - microparticles - integrins

Supplementary Material

 

  • References

  • 1 van der Veen BS, de Winther MP, Heeringa P. Myeloperoxidase: molecular mechanisms of action and their relevance to human health and disease. Antioxid Redox Signal 2009; 11 (11) 2899-2937
    CrossrefPubMedGoogle Scholar
  • 2 Davies MJ. Myeloperoxidase-derived oxidation: mechanisms of biological damage and its prevention. J Clin Biochem Nutr 2011; 48 (01) 8-19
    CrossrefPubMedGoogle Scholar
  • 3 Kim H, Wei Y, Lee JY. , et al. Myeloperoxidase inhibition increases neurogenesis after ischemic stroke. J Pharmacol Exp Ther 2016; 359 (02) 262-272
    CrossrefPubMedGoogle Scholar
  • 4 Teng N, Maghzal GJ, Talib J, Rashid I, Lau AK, Stocker R. The roles of myeloperoxidase in coronary artery disease and its potential implication in plaque rupture. Redox Rep 2017; 22 (02) 51-73
    CrossrefPubMedGoogle Scholar
  • 5 Rudolph V, Andrié RP, Rudolph TK. , et al. Myeloperoxidase acts as a profibrotic mediator of atrial fibrillation. Nat Med 2010; 16 (04) 470-474
    CrossrefPubMedGoogle Scholar
  • 6 Fuchs TA, Brill A, Wagner DD. Neutrophil extracellular trap (NET) impact on deep vein thrombosis. Arterioscler Thromb Vasc Biol 2012; 32 (08) 1777-1783
    CrossrefPubMedGoogle Scholar
  • 7 Pfeiler S, Stark K, Massberg S, Engelmann B. Propagation of thrombosis by neutrophils and extracellular nucleosome networks. Haematologica 2017; 102 (02) 206-213
    CrossrefPubMedGoogle Scholar
  • 8 Massberg S, Grahl L, von Bruehl ML. , et al. Reciprocal coupling of coagulation and innate immunity via neutrophil serine proteases. Nat Med 2010; 16 (08) 887-896
    CrossrefPubMedGoogle Scholar
  • 9 Fuchs TA, Brill A, Duerschmied D. , et al. Extracellular DNA traps promote thrombosis. Proc Natl Acad Sci U S A 2010; 107 (36) 15880-15885
    CrossrefPubMedGoogle Scholar
  • 10 von Brühl ML, Stark K, Steinhart A. , et al. Monocytes, neutrophils, and platelets cooperate to initiate and propagate venous thrombosis in mice in vivo. J Exp Med 2012; 209 (04) 819-835
    CrossrefPubMedGoogle Scholar
  • 11 Fuchs TA, Kremer Hovinga JA, Schatzberg D, Wagner DD, Lämmle B. Circulating DNA and myeloperoxidase indicate disease activity in patients with thrombotic microangiopathies. Blood 2012; 120 (06) 1157-1164
    CrossrefPubMedGoogle Scholar
  • 12 Metzler KD, Fuchs TA, Nauseef WM. , et al. Myeloperoxidase is required for neutrophil extracellular trap formation: implications for innate immunity. Blood 2011; 117 (03) 953-959
    CrossrefPubMedGoogle Scholar
  • 13 Subramaniam S, Jurk K, Hobohm L. , et al. Distinct contributions of complement factors to platelet activation and fibrin formation in venous thrombus development. Blood 2017; 129 (16) 2291-2302
    CrossrefPubMedGoogle Scholar
  • 14 Banerjee S, Stampler J, Furtmüller PG, Obinger C. Conformational and thermal stability of mature dimeric human myeloperoxidase and a recombinant monomeric form from CHO cells. Biochim Biophys Acta 2011; 1814 (02) 375-387
    CrossrefPubMedGoogle Scholar
  • 15 Lyberg T, Prydz H. Phorbol esters induce synthesis of thromboplastin activity in human monocytes. Biochem J 1981; 194 (03) 699-706
    CrossrefPubMedGoogle Scholar
  • 16 Noubouossie DF, Whelihan MF, Yu YB. , et al. In vitro activation of coagulation by human neutrophil DNA and histone proteins but not neutrophil extracellular traps. Blood 2017; 129 (08) 1021-1029
    CrossrefPubMedGoogle Scholar
  • 17 Shi J, Shi Y, Waehrens LN, Rasmussen JT, Heegaard CW, Gilbert GE. Lactadherin detects early phosphatidylserine exposure on immortalized leukemia cells undergoing programmed cell death. Cytometry A 2006; 69 (12) 1193-1201
    PubMedGoogle Scholar
  • 18 Topalov NN, Yakimenko AO, Canault M. , et al. Two types of procoagulant platelets are formed upon physiological activation and are controlled by integrin α(IIb)β(3). Arterioscler Thromb Vasc Biol 2012; 32 (10) 2475-2483
    CrossrefPubMedGoogle Scholar
  • 19 Rochat S, Alberio L. Formaldehyde-fixation of platelets for flow cytometric measurement of phosphatidylserine exposure is feasible. Cytometry A 2015; 87 (01) 32-36
    CrossrefPubMedGoogle Scholar
  • 20 Johansson MW, Patarroyo M, Oberg F, Siegbahn A, Nilsson K. Myeloperoxidase mediates cell adhesion via the alpha M beta 2 integrin (Mac-1, CD11b/CD18). J Cell Sci 1997; 110 (Pt 9): 1133-1139
    PubMedGoogle Scholar
  • 21 Lau D, Mollnau H, Eiserich JP. , et al. Myeloperoxidase mediates neutrophil activation by association with CD11b/CD18 integrins. Proc Natl Acad Sci U S A 2005; 102 (02) 431-436
    CrossrefPubMedGoogle Scholar
  • 22 El Kebir D, József L, Pan W, Filep JG. Myeloperoxidase delays neutrophil apoptosis through CD11b/CD18 integrins and prolongs inflammation. Circ Res 2008; 103 (04) 352-359
    CrossrefPubMedGoogle Scholar
  • 23 Flemmig J, Lessig J, Reibetanz U, Dautel P, Arnhold J. Non-vital polymorphonuclear leukocytes express myeloperoxidase on their surface. Cell Physiol Biochem 2008; 21 (04) 287-296
    CrossrefPubMedGoogle Scholar
  • 24 Lessig J, Spalteholz H, Reibetanz U. , et al. Myeloperoxidase binds to non-vital spermatozoa on phosphatidylserine epitopes. Apoptosis 2007; 12 (10) 1803-1812
    CrossrefPubMedGoogle Scholar
  • 25 Flemmig J, Spalteholz H, Schubert K, Meier S, Arnhold J. Modification of phosphatidylserine by hypochlorous acid. Chem Phys Lipids 2009; 161 (01) 44-50
    CrossrefPubMedGoogle Scholar
  • 26 Flemmig J, Arnhold J. Interaction of hypochlorous acid and myeloperoxidase with phosphatidylserine in the presence of ammonium ions. J Inorg Biochem 2010; 104 (07) 759-764
    CrossrefPubMedGoogle Scholar
  • 27 Bach RR. Tissue factor encryption. Arterioscler Thromb Vasc Biol 2006; 26 (03) 456-461
    PubMedGoogle Scholar
  • 28 Langer F, Ruf W. Synergies of phosphatidylserine and protein disulfide isomerase in tissue factor activation. Thromb Haemost 2014; 111 (04) 590-597
    Article in Thieme ConnectPubMedGoogle Scholar
  • 29 Dicke C, Amirkhosravi A, Spath B. , et al. Tissue factor-dependent and -independent pathways of systemic coagulation activation in acute myeloid leukemia: a single-center cohort study. Exp Hematol Oncol 2015; 4: 22
    CrossrefPubMedGoogle Scholar
  • 30 Lewis HD, Liddle J, Coote JE. , et al. Inhibition of PAD4 activity is sufficient to disrupt mouse and human NET formation. Nat Chem Biol 2015; 11 (03) 189-191
    CrossrefPubMedGoogle Scholar
  • 31 Kubala L, Lu G, Baldus S, Berglund L, Eiserich JP. Plasma levels of myeloperoxidase are not elevated in patients with stable coronary artery disease. Clin Chim Acta 2008; 394 (1-2): 59-62
    CrossrefPubMedGoogle Scholar
  • 32 Heslop CL, Frohlich JJ, Hill JS. Myeloperoxidase and C-reactive protein have combined utility for long-term prediction of cardiovascular mortality after coronary angiography. J Am Coll Cardiol 2010; 55 (11) 1102-1109
    CrossrefPubMedGoogle Scholar
  • 33 Tzikas S, Schlak D, Sopova K. , et al. Increased myeloperoxidase plasma levels in patients with Alzheimer's disease. J Alzheimers Dis 2014; 39 (03) 557-564
    CrossrefPubMedGoogle Scholar
  • 34 Mayyas FA, Al-Jarrah MI, Ibrahim KS, Alzoubi KH. Level and significance of plasma myeloperoxidase and the neutrophil to lymphocyte ratio in patients with coronary artery disease. Exp Ther Med 2014; 8 (06) 1951-1957
    CrossrefPubMedGoogle Scholar
  • 35 Nakamuta M, Ohashi M, Tanabe Y, Hiroshige K, Nawata H. High plasma concentration of myeloperoxidase in cirrhosis: a possible marker of hypersplenism. Hepatology 1993; 18 (06) 1377-1383
    CrossrefPubMedGoogle Scholar
  • 36 d'Onofrio G, Mancini R, Vallone R. , et al. Acquired neutrophil myeloperoxidase deficiency: an indicator of subclinical activation of blood coagulation?. Blood Cells 1983; 9 (03) 455-466
    PubMedGoogle Scholar
  • 37 Kalinski T, Jentsch-Ullrich K, Fill S, König B, Costa SD, Roessner A. Lethal candida sepsis associated with myeloperoxidase deficiency and pre-eclampsia. APMIS 2007; 115 (07) 875-880
    CrossrefPubMedGoogle Scholar
  • 38 Kutter D. Prevalence of myeloperoxidase deficiency: population studies using Bayer-Technicon automated hematology. J Mol Med (Berl) 1998; 76 (10) 669-675
    CrossrefPubMedGoogle Scholar
  • 39 Campbell PJ, Green AR. The myeloproliferative disorders. N Engl J Med 2006; 355 (23) 2452-2466
    CrossrefPubMedGoogle Scholar
  • 40 Falanga A, Marchetti M. Thrombotic disease in the myeloproliferative neoplasms. Hematology (Am Soc Hematol Educ Program) 2012; 2012: 571-581
    PubMedGoogle Scholar