Procoagulant activity and phosphatidylserine of amniotic fluid cells

Jin Zhou; Shuchuan Liu; Ming Ma; Jinxiao Hou; Hongjuan Yu; Chengfang Lu; Gary E. Gilbert; Jialan Shi

doi:10.1160/TH08-08-0531

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2009; 101(05): 845-851
DOI: 10.1160/TH08-08-0531

Blood Coagulation, Fibrinolysis and Cellular Haemostasis

Schattauer GmbH

Procoagulant activity and phosphatidylserine of amniotic fluid cells

Authors

Jin Zhou

¹Department of Hematology, the First Affiliated Hospital, Harbin Medical University, Harbin, China
Shuchuan Liu

¹Department of Hematology, the First Affiliated Hospital, Harbin Medical University, Harbin, China
Ming Ma

¹Department of Hematology, the First Affiliated Hospital, Harbin Medical University, Harbin, China
Jinxiao Hou

¹Department of Hematology, the First Affiliated Hospital, Harbin Medical University, Harbin, China
Hongjuan Yu

¹Department of Hematology, the First Affiliated Hospital, Harbin Medical University, Harbin, China
Chengfang Lu

¹Department of Hematology, the First Affiliated Hospital, Harbin Medical University, Harbin, China
Gary E. Gilbert

²Department of Medicine, VA Boston Health Care System, Brigham and Women’s Hospital and Harvard Medical School, Boston, Massachusetts, USA
Jialan Shi

¹Department of Hematology, the First Affiliated Hospital, Harbin Medical University, Harbin, China

²Department of Medicine, VA Boston Health Care System, Brigham and Women’s Hospital and Harvard Medical School, Boston, Massachusetts, USA

Abstract

Summary

Amniotic fluid (AF) may induce disseminated intravascular coagulation (DIC) when it enters maternal circulation by breaching the placental-maternal circulation barrier. The precise mechanism of the procoagulant activity of AF is unclear, but tissue factor (TF) has been proposed to be the main cause. As one constituent of AF, AF cells accumulate and undergo apoptosis continuously. Therefore, we speculate that AF cells have procoagulant activity due to the externalisation of phosphatidylserine (PS). The present study aims to demonstrate that, in addition to TF, the PS that is externalised on AF cells is important for the procoagulant activity of AF. Ten AF samples from parturient women were analysed using lactadherin as the probe for PS. Anti-TF antibody also was used to identify TF and its associated coagulation functions in AF cells. Normal platelets, neutrophils, and lymphocytes were harvested as controls. Confocal microscopy and flow cytometry was used to assess PS expression on AF cells. The procoagulant activity of AF cells was demonstrated by a plasma coagulation assay and further confirmed by factor Xase/prothrombinase assays. PS and TF were present on most AF cells, providing substantial procoagulant activity. Furthermore, factor Xase and prothrombinase assays showed that AF cells substantially enforced the activation of factor X and prothrombin. PS on AF cells is an important procoagulant source for AF. Lactadherin is an ideal anticoagulant for inhibiting the procoagulant activity of AF cells.

Keywords

Amniotic fluid cells - phosphatidylserine - procoagulant activity - lactadherin

Full Text

References

References
1 Benson MD. A hypothesis regarding complement activation and amniotic fluid embolism. Med Hypotheses 2007; 68: 1019-1025.
2 Phillips LL, Davidson EC. Procoagulant properties of amniotic fluid. Am J Obstet Gynecol 1972; 113: 911-919.
3 Clark SL, Hankins GD, Dudley DA. et al. Amniotic fluid embolism: Analysis of the national registry. Am J Obstet Gynecol 1995; 172: 1158-1167.
4 Harnett MJ, Hepner DL, Datta S. et al. Effect of amniotic fluid on coagulation and platelet function in pregnancy: An evaluation using thromboelastography. Anaesthesia 2005; 60: 1068-1072.
5 Awad IT, Shorten GD. Amniotic fluid embolism and isolated coagulopathy: Atypical presentation of amniotic fluid embolism. Eur J Anaesthesiol 2001; 18: 410-413.
6 Tydén O, Bergström S, Nilsson BA. Origin of amniotic cells in mid-tremester pregnancies. Br J Obstet Gynaecol 1981; 88: 278-286.
7 Bombeli T, Karsan A, Tait JF. et al. Apoptotic vascular endothelial cells become procoagulant. Blood 1997; 89: 2429-2442.
8 Solum NO. Procoagulant expression in platelets and defects leading to clinical disorders. Arterioscler Thromb Vasc Biol 1999; 19: 2841-2846.
9 Vance JE, Steenbergen R. Metabolism and functions of phosphatidylserine. Proq Lipid Res 2005; 44: 207-234.
10 Østerud B, Bjorklid E. The tissue factor pathway in disseminated intravascular coagulation. Semin Thromb Hemost 2001; 27: 605-617.
11 Stubbs JD, Lekutis C, Singer KL. et al. cDNA cloning of a mouse mammary epithelial cell surface protein reveals the existence of epidermal growth factor-like domains linked to factor VIII-like sequences. Proc Natl Acad Sci USA 1990; 87: 8417-8421.
12 Shi J, Heegaard CW, Rasmussen JT. et al. Lactadherin binds selectively to membranes containing phosphatidyl-L-serine and increased curvature. Biochim Biophys Acta 2004; 1667: 82-90.
13 Shi J, Gilbert GE. Lactadherin inhibits enzyme complexes of blood coagulation by competing for phospholipid-binding sites. Blood 2003; 101: 2628-2636.
14 Dasgupta SK, Guchhait P, Thiagarajan P. Lactadherin binding and phosphatidylserine expression on cell surface-comparison with annexin A5. Transl Res 2006; 148: 19-25.
15 Hvarregaard J, Andersen MH, Berglund L. et al. Characterization of glycoprotein PAS-6/7 from membranes of bovine milk fat globules. Eur J Biochem 1996; 240: 628-636.
16 Kim DH, Kanno C, Mizokami Y. Purification and characterization of major glycoproteins, PAS-6 and PAS-7, from bovine milk fat globule membrane. Biochim Biophys Acta 1992; 1122: 203-211.
17 Uszyzski M, Zekanowska E, Uszyzski W. et al. Tissue factor (TF) and tissue factor pathway inhibitor (TFPI) in amniotic fluid and blood plasma: implications for the mechanism of amniotic fluid embolism. Eur J Obstet Gynecol Reprod Biol 2001; 95: 163-166.
18 Kumar D, Fung W, Moore RM. et al. Proinflamma-tory cytokines found in amniotic fluid induce collagen remodeling, apoptosis, and biophysical weakening of cultured human fetal membranes. Biol Reprod 2006; 74: 29-34.
19 Bombeli T, Karsan A, Tait JF. et al. Apoptotic vascular endothelial cells become procoagulant. Blood 1997; 89: 2429-2442.
20 Gilbert GE, Furie BC, Furie B. Binding of human factor VIII to phospholipid vesicles. J Biol Chem 1990; 265: 815-822.
21 Gilbert GE, Arena AA. Activation of the factor VIIIa-factor IXa enzyme complex of blood coagulation by membranes containing phosphatidyl-L-serine. J Biol Chem 1996; 271: 11120-11125.
22 Majumder R, Weinreb G, Lentz BR. Efficient thrombin generation requires molecular phosphatidylserine, not a membrane surface. Biochemistry 2005; 44: 16998-17006.
23 Majumder R, Quinn-Allen MA, Kane WH. et al. The phosphatidylserine binding site of the factor Va C2 domain accounts for membrane binding but does not contribute to the assembly or activity of a human factor Xa-factor Va complex. Biochemistry 2005; 44: 711-718.
24 Gale AJ, Yegneswaran S, Xu X. et al. Characterization of a factor Xa binding site on factor Va near the Arg-506 activated protein C cleavage site. J Biol Chem 2007; 282: 21848-21855.
25 Majumder R, Weinreb G, Zhai X. et al. Soluble phosphatidylserine triggers assembly in solution of a prothrombin-activating complex in the absence of a membrane surface. J Biol Chem 2002; 277: 29765-29773.
26 Wang J, Weiss I, Svoboda K. et al. Thrombogenic role of cells undergoing apoptosis. Br J Haemotol 2001; 115: 382-391.
27 Lockwood CJ, Bach R, Guha A. et al. Amniotic fluid contains tissue factor, a potent initiator of coagulation. Am J Obstet Gynecol 1991; 165: 1335-1341.
28 Fujii M, Hayakawa H, Urano T. et al. Relevance of tissue factor and tissue factor pathway inhibitor for hypercoagulable state in the lungs of patients with idiopathic pulmonary fibrosis. Thromb Res 2000; 99: 111-117.
29 Lurie S, Feinstein M, Mamet Y. Disseminated intravascular coagulopathy in pregnancy: Thorough comprehension of etiology and management reduces obstetricians’ stress. Arch Gynecol Obstet 2000; 263: 126-130.
30 Uszyzski W, Uszyzski M, Zekanowska E. Thrombin activatable fibrinolysis inhibitor (TAFI) in human amniotic fluid. A preliminary study. Thromb Res 2007; 119: 241-245.
31 Uszyzski M, Zekanowska E, Kotzbach M. et al. Protein C, protein S, and thrombomodulin in amniotic fluid. A preliminary study. J Perinat Med 2006; 34: 289-292.