Leukocyte- and platelet-derived microparticles correlate with thrombus weight and tissue factor activity in an experimental mouse model of venous thrombosis

 

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Summary

Microparticles (MP) are lipid vesicles from platelets, leukocytes and endothelial cells that are involved in early thrombogenesis. We evaluated a detailed time-course analysis of MPs on thrombogenesis and the associated tissue factor (TF) activity in wild-type, in gene-deleted for E- and P-selectins and with high levels of P-selectin expression after the initiation of venous thrombosis in mice.Inferior vena cava (IVC) ligation was performed on C57BL/6 mice (n =191, 59 = wild-type [WT], 55 = gene-deleted for E- and P – selectins [knock-outs, EPKO] and 77 = elevated levels of soluble P-selectin, named Delta Cytoplasmic Tail (ΔCT). Animals were euthanised at various time points to assess MP production, origin and thrombus weight. MPs were re-injected into separate mice at concentrations of 80,000 and 160,000 units, as well as from different ages. In addition, MPs from thrombosed animals were pooled and TF activity quantitated using a chromogenic assay. Thrombus weight correlated negatively with MPs derived from leukocytes, and positively with MPs derived from platelets for WT animals (p<0.05), while MPs from platelets presented a positive correlation to thrombus weight in the WT and EPKO groups (p<0.01). Total MPs correlated negatively with thrombus weight in the ΔCT group (p<0.05). MP re-injections led to greater thrombus weight, while older MP reinjections tended to form larger thrombus than younger. Finally, TF bearing MPs showed a significant correlation to MP concentrations (R=0.99). In conclusion, MPs appear to be an important element in venous thrombogenesis.

• References

• 1 Stewart GJ, Ritchie WGM, Lynch PR. Venous endothelial damage produced by massive sticking and emigration of leukocytes. Am J Pathol 1974; 74: 507-532.
  PubMedGoogle Scholar
• 2 Virchow R. Gesammelte Abhandlungen zur Wissenschaftlichen Medizin. Frankfurt: A M Von Meidinger Sohn; 1856: 525-530.
  Google Scholar
• 3 Esmon CT. Inflammation and thrombosis. J Thromb Haemost 2003; 1: 1343-1348.
  CrossrefPubMedGoogle Scholar
• 4 Myers DD, Hawley AE, Farris DM. et al. P-selectin and leukocyte microparticles are associated with venous thrombogenesis. J Vasc Surg 2003; 38: 1075-1089.
  CrossrefPubMedGoogle Scholar
• 5 Gilbert GE, Sims PJ, Wiedmer T. et al. Platelet-derived MPs express high affinity receptors for factor VIII. J Biol Chem 1991; 266: 17261-17268.
  PubMedGoogle Scholar
• 6 Mesri M, Altieri DC. Endothelial cell activation by leukocyte MPs. J Immunol 1998; 161: 4382-4387.
  PubMedGoogle Scholar
• 7 Sabatier F, Roux V, Anfosso F. et al. Interaction of endothelial MPs with monocytic cells in vitro induces tissue factor-dependent procoagulant activity. Blood 2002; 99: 3962-3970.
  CrossrefPubMedGoogle Scholar
• 8 Deregibus MC, Cantaluppi V, Calogero R. et al. Endothelial progenitor cell derived microvesicles activate an angiogenic program in endothelial cells by a horizontal transfer of mRNA. Blood 2007; 110: 2440-2448.
  CrossrefPubMedGoogle Scholar
• 9 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.
  CrossrefPubMedGoogle Scholar
• 10 Martinez MC, Tesse A, Zobairi F. et al. Shed membrane MPs from circulating and vascular cells in regulating vascular function. Am J Physiol Heart Circ Physiol 2005; 288: H1004-H1009.
  CrossrefPubMedGoogle Scholar
• 11 Sevinsky JR, Rao LV, Ruf W. Ligand-induced pro-tease receptor translocation into caveolae: a mechanism for regulating cell surface proteolysis of the tissue factor-dependent coagulation pathway. J Cell Biol 1996; 133: 293-304.
  CrossrefPubMedGoogle Scholar
• 12 Bullard DC, Qin L, Lorenzo I. et al. P-Selectin/ ICAM-1 double mutant mice: acute emigration of neutrophils into the peritoneum is completely absent but is normal into pulmonary alveoli. J Clin Invest 1995; 95: 1782-1788.
  CrossrefPubMedGoogle Scholar
• 13 Bullard DC, Kunkel EJ, Kubo H. et al. Infectious susceptibility and severe deficiency of leukocyte rolling and recruitment in E-selectin and P-selectin double mutant mice. J Exp Med 1996; 183: 2329-2336.
  CrossrefPubMedGoogle Scholar
• 14 Andre P, Hartwell D, Hrachovinova I. et al. Pro-coagulant state resulting from high levels of soluble P-selectin in blood. Proc Natl Acad Sci USA 2000; 97: 13835-13840.
  CrossrefPubMedGoogle Scholar
• 15 Hartwell DW, Mayadas TN, Berger G. et al. Role of P-selectin cytoplasmic domain in granular targeting in vivo and in early inflammatory responses. J Cell Biol 1998; 143: 1129-1141.
  CrossrefPubMedGoogle Scholar
• 16 Osterud B. The role of platelets in decrypting monocyte tissue factor. Semin Hematol 2001; 38 4 Suppl 12 2-5.
  PubMedGoogle Scholar
• 17 Myers Jr DD, Rectenwald JE. et al. Decreased venous thrombosis with an oral inhibitor of P- selectin. J Vasc Surg 2005; 42: 329-336.
  CrossrefPubMedGoogle Scholar
• 18 Myers DD, Wakefield TW. Inflammation-dependent thrombosis. Front Biosci 2005; Sep 1 10: 2750-2757.
  CrossrefPubMedGoogle Scholar
• 19 Polgar J, Matuskova J, Wagner DD. The P-selectin, tissue factor, coagulation triad. J Thromb Haemost 2005; 3: 1590-1596.
  CrossrefPubMedGoogle Scholar
• 20 Giesen PL, Rauch U, Bohrmann B. et al. Blood-borne tissue factor: another view of thrombosis. Proc Natl Acad Sci USA 1999; 96: 2311-2315.
  CrossrefPubMedGoogle Scholar
• 21 Jy W, Horstman LL, Wang F. et al. Platelet factor 3 in plasma fractions: its relation to microparticle size and thromboses. Thromb Res 1995; 80: 471-482.
  CrossrefPubMedGoogle Scholar
• 22 Hrachovinová I, Cambien B, Hafezi-Moghadam A. et al. Interaction of P-selectin and PSGL-1 generates MPs that correct hemostasis in a mouse model of hemophilia A. Nat Med 2003; 9: 1020-1025.
  CrossrefPubMedGoogle Scholar
• 23 Biró E, Sturk-Maquelin KN, Vogel GM. et al. Human cell-derived MPs promote thrombus formation in vivo in a tissue factor-dependent manner. J Thromb Haemost 2003; 1: 2561-2568.
  CrossrefPubMedGoogle Scholar
• 24 Podor TJ, Singh D, Chindemi P. et al. Vimentin exposed on activated platelets and platelet MPs localizes vitronectin and plasminogen activator inhibitor complexes on their surface. J Biol Chem 2002; 277: 7529-7539.
  CrossrefPubMedGoogle Scholar
• 25 Wakefield TW, Myers DD, Henke PK. Mechanisms of venous thrombosis and resolution. Arterioscler Thromb Vasc Biol 2008; 28: 387-391.
  CrossrefPubMedGoogle Scholar
• 26 Rectenwald JE, Myers Jr DD, Hawley AE. et al. D-dimer, P-selectin, and MPs: novel markers to predict deep venous thrombosis. A pilot study. Thromb Haemost 2005; 94: 1312-1317.
  Article in Thieme ConnectPubMedGoogle Scholar
• 27 Chirinos JA, Heresi GA, Velasquez H. et al. Elevation of endothelial MPs, platelets, and leukocyte activation in patients with venous thromboembolism. J Am Coll Cardiol 2005; 45: 1467-1471.
  CrossrefPubMedGoogle Scholar
• 28 Rauch U, Nemerson Y. Circulating tissue factor and thrombosis. Current Opin Hematol 2000; 7: 273-277.
  PubMedGoogle Scholar
• 29 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.
  CrossrefPubMedGoogle Scholar