Discrepancy between tissue factor activity and tissue factor expression in endotoxin-induced monocytes is associated with apoptosis and necrosis

Carola E. Henriksson; Olav Klingenberg; Reidun Øvstebø; Gun-Britt Joø; Åse-Brit Westvik; Peter Kierulf

doi:10.1160/TH05-07-0463

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2005; 94(06): 1236-1244
DOI: 10.1160/TH05-07-0463

Platelets and Blood Cells

Schattauer GmbH

Discrepancy between tissue factor activity and tissue factor expression in endotoxin-induced monocytes is associated with apoptosis and necrosis

Carola E. Henriksson

¹The R&D Group, Department of Clinical Chemistry, Ullevaal University Hospital, Oslo, Norway

,

Olav Klingenberg

¹The R&D Group, Department of Clinical Chemistry, Ullevaal University Hospital, Oslo, Norway

,

Reidun Øvstebø

¹The R&D Group, Department of Clinical Chemistry, Ullevaal University Hospital, Oslo, Norway

,

Gun-Britt Joø

¹The R&D Group, Department of Clinical Chemistry, Ullevaal University Hospital, Oslo, Norway

,

Åse-Brit Westvik

¹The R&D Group, Department of Clinical Chemistry, Ullevaal University Hospital, Oslo, Norway

,

Peter Kierulf

¹The R&D Group, Department of Clinical Chemistry, Ullevaal University Hospital, Oslo, Norway

› Author Affiliations

Abstract

Summary

Tissue factor (TF), the main initiator of blood coagulation, contributes to the manifestation of disseminated intravascular coagulation following septic shock in meningococcal infection. Since a direct relationship between disease severity and lipopolysaccharide (LPS) concentration in the circulation has been shown, we hypothesized that the procoagulant and cytotoxic effects of endotoxin also in vitro were related to its concentration. In vitro studies, however, have frequently used much higher LPS concentrations than those observed in clinical samples. Using elutriation-purified human monocytes, we observed that LPS up to 1000 ng/ml exerted a concentration-dependent increase in TF activity (tenase activity, fibrin formation in plasma). Although there was a dose-dependent increase in TF activity, there was not a concomitant increase in TF expression at LPS concentrations above 1 ng/ml (flow cytometry, Western blotting, TF mRNA). Flow cytometry revealed that this discrepancy between TF activity and TF expression at endotoxin concentrations above 1 ng/ml, coincided with an LPS dose-dependent increase in cell surface phosphatidylserine (PS), considered to promote coagulation. The increased PS expression was associated with an increased number of 7-AAD-positive cells indicating cell death. We conclude that enhancement of monocyte procoagulant activity in vitro by high concentrations of LPS may result from increased PS exposure due to apoptosis and necrosis. Therefore, the LPS concentrations used to examine monocyte procoagulant activity in vitro, should be carefully chosen.

Keywords

Tissue factor - coagulation - lipopolysaccharide - phosphatidylserine - monocyte

Full Text

References

References
1 Osterud B, Flaegstad T. Increased tissue thromboplastin activity in monocytes of patients with meningococcal infection: related to an unfavourable prognosis. Thromb Haemost 1983; 49: 5-7.
2 Osterud B. A global view on the role of monocytes and platelets in atherogenesis. Thromb Res 1997; 85: 1-22.
3 Huang DR, Wang J, Kivisakk P. et al Absence of monocyte chemoattractant protein 1 in mice leads to decreased local macrophage recruitment and antigenspecific T helper cell type 1 immune response in experimental autoimmune encephalomyelitis. J Exp Med 2001; 193: 713-26.
4 Prydz H, Lyberg T, Deteix P. et al In vitro stimulation of tissue thromboplastin (factor III) activity in human monocytes by immune complexes and lectins. Thromb Res 1979; 15: 465-74.
5 Schwartz BS, Levy GA, Curtiss LK. et al Plasma lipoprotein induction and suppression of the generation of cellular procoagulant activity in vitro: two procoagulant activities are produced by peripheral blood mononuclear cells. J Clin Invest 1981; 67: 1650-8.
6 Broze Jr. GJ. Binding of human factor VII and VIIa to monocytes. J Clin Invest 1982; 70: 526-35.
7 Rapaport SI, Rao LV. Initiation and regulation of tissue factor-dependent blood coagulation. Arterioscler Thromb 1992; 12: 1111-21.
8 Roberts HR, Lozier JN. New perspectives on the coagulation cascade. Hosp Pract (Off Ed) 1992; 27: 97-12.
9 Wolberg AS, Monroe DM, Roberts HR. et al Tissue factor de-encryption: ionophore treatment induces changes in tissue factor activity by phosphatidylserinedependent and -independent mechanisms. Blood Coagul Fibrinolysis 1999; 10: 201-10.
10 Aupeix K, Toti F, Satta N. et al Oyxsterols induce membrane procoagulant activity in monocytic THP-1 cells. Biochem J 1996; 314 (Pt 3) 1027-33.
11 Greeno EW, Bach RR, Moldow CF. Apoptosis is associated with increased cell surface tissue factor procoagulant activity. Lab Invest 1996; 75: 281-9.
12 Mann KG, Nesheim ME, Church WR. et al Surface-dependent reactions of the vitamin K-dependent enzyme complexes. Blood 1990; 76: 1-16.
13 Ruf W, Rehemtulla A, Morrissey JH. et al Phospholipid-independent and -dependent interactions required for tissue factor receptor and cofactor function. J Biol Chem 1991; 266: 16256.
14 Ruf W, Edgington TS. Structural biology of tissue factor, the initiator of thrombogenesis in vivo . FASEB J 1994; 8: 385-90.
15 Bach RR, Moldow CF. Mechanism of tissue factor activation on HL-60 cells. Blood 1997; 89: 3270-6.
16 Bruckheimer EM, Schroit AJ. Membrane phospholipid asymmetry: host response to the externalization of phosphatidylserine. J Leukoc Biol 1996; 59: 784-8.
17 Bach R, Rifkin DB. Expression of tissue factor procoagulant activity: regulation by cytosolic calcium. Proc Natl Acad Sci U S A 1990; 87: 6995-9.
18 Barrowcliffe TW, Fabregas P, Jardi M. et al Procoagulant activity of T lymphoblastoid cells due to exposure of negatively charged phospholipid.[see comment]. Thromb Haemost 2002; 87: 442-9.
19 Setty BN, Kulkarni S, Rao AK. et al Fetal hemoglobin in sickle cell disease: relationship to erythrocyte phosphatidylserine exposure and coagulation activation. Blood 2000; 96: 1119-24.
20 Gregory SA, Morrissey JH, Edgington TS. Regulation of tissue factor gene expression in the monocyte procoagulant response to endotoxin. Mol Cell Biol 1989; 9: 2752-5.
21 Nemerson Y. Tissue factor and hemostasis. Blood 1988; 71: 1-8.
22 Osnes LT, Westvik AB, Kierulf P. Procoagulant and profibrinolytic activities of cryopreserved human monocytes. Thromb Res 1994; 76: 373-83.
23 Robinson RA, Worfolk L, Tracy PB. Endotoxin enhances the expression of monocyte prothrombinase activity. Blood 1992; 79: 406-16.
24 Karahashi H, Amano F. Lipopolysaccharide (LPS)-induced cell death of C3H mouse peritoneal macrophages in the presence of cycloheximide: different susceptibilities of C3H/HeN and C3H/HeJ mice macrophages. J Endotoxin Res 2000; 6: 33-9.
25 Brandtzaeg P, Kierulf P, Gaustad P. et al Plasma endotoxin as a predictor of multiple organ failure and death in systemic meningococcal disease. J Infect Dis 1989; 159: 195-204.
26 Satta N, Toti F, Feugeas O. et al Monocyte vesiculation is a possible mechanism for dissemination of membrane-associated procoagulant activities and adhesion molecules after stimulation by lipopolysaccharide. J Immunol 1994; 153: 3245-55.
27 Wharton SA, Riley PA. The suppression of endogenous protein degradation by fractions of foetal calf serum: dialysed serum is less able to suppress degradation in aged cells. Cell Biochem Funct 1986; 4: 189-95.
28 Lund PK, Joo GB, Westvik AB. et al Isolation of monocytes from whole blood by density gradient centrifugation and counter-current elutriation followed by cryopreservation: six years’ experience. Scand J Clin Lab Invest 2000; 60: 357-65.
29 Nossel HL, Younger LR, Wilner GD. et al Radioimmunoassay of human fibrinopeptide A. Proc Natl Acad Sci U S A 1971; 68: 2350-3.
30 Kockum C, Frebelius S. Rapid radioimmunoassay of human fibrinopeptide A – removal of cross-reacting fibrinogen with bentonite. Thromb Res 1980; 19: 589-98.
31 Osnes LT, Haug KB, Joo GB. et al Aspirin potentiates LPS-induced fibrin formation (FPA) and TNFalpha-synthesis in whole blood. Thromb Haemost 2000; 83: 868-73.
32 Ovstebo R, Haug KB, Lande K. et al PCR-based calibration curves for studies of quantitative gene expression in human monocytes: development and evaluation. Clin Chem 2003; 49: 425-32.
33 Ollivier V, Houssaye S, Ternisien C. et al Endotoxin-induced tissue factor messenger RNA in human monocytes is negatively regulated by a cyclic AMP-dependent mechanism. Blood 1993; 81: 973-9.
34 Krishnaswamy S, Field KA, Edgington TS. et al Role of the membrane surface in the activation of human coagulation factor X. J Biol Chem 1992; 267: 26110-20.
35 Le DT, Rapaport SI, Rao LV. Relations between factor VIIa binding and expression of factor VIIa/tissue factor catalytic activity on cell surfaces. J Biol Chem 1992; 267: 15447-54.
36 Hansen CB, van Deurs B, Petersen LC. et al Discordant expression of tissue factor and its activity in polarized epithelial cells. Asymmetry in anionic phospholipid availability as a possible explanation. Blood 1999; 94: 1657-64.
37 Donate F, Kelly CR, Ruf W. et al Dimerization of tissue factor supports solution-phase autoactivation of factor VII without influencing proteolytic activation of factor X. Biochemistry (Mosc) 2000; 39: 11467-76.
38 Freyssinet JM. Cellular microparticles: what are they bad or good for?. J Thromb Haemost 2003; 1: 1655-62.
39 VanWijk MJ, VanBavel E, Sturk A. et al Microparticles in cardiovascular diseases. Cardiovasc Res 2003; 59: 277-87.
40 Nieuwland R, Berckmans RJ, McGregor S. et al Cellular origin and procoagulant properties of microparticles in meningococcal sepsis. Blood 2000; 95: 930-5.
41 Mallat Z, Hugel B, Ohan J. et al Shed membrane microparticles with procoagulant potential in human atherosclerotic plaques: a role for apoptosis in plaque thrombogenicity. Circulation 1999; 99: 348-53.
42 Mallat Z, Benamer H, Hugel B. et al Elevated levels of shed membrane microparticles with procoagulant potential in the peripheral circulating blood of patients with acute coronary syndromes. Circulation 2000; 101: 841-3.
43 Dietzen DJ, Page KL, Tetzloff TA. Lipid rafts are necessary for tonic inhibition of cellular tissue factor procoagulant activity. Blood 2004; 103: 3038-44.
44 Kunzelmann-Marche C, Freyssinet JM, Martinez MC. Loss of plasma membrane phospholipid asymmetry requires raft integrity. Role of transient receptor potential channels and ERK pathway. J Biol Chem 2002; 277: 19876-81.