Pre-analytical and analytical issues in the analysis of blood microparticles

Summary

Results of plasma microparticles (MPs) measurements reported in the literature vary widely. This is clearly not only related to the lack of well-standardised MP assays, but also to variations in pre-analytical conditions. In this review we will discuss the pre-analytical variables related to plasma and MP preparation which may affect MP analysis. Additionally we will address several analytical issues in commonly used MP as-says and briefly discuss some novel approaches for the detection and characterisation of MPs. Ideally MP measurements should be performed in plasma, freshly prepared directly after blood withdrawal. As platelet contamination seems to be one of the major pre-analytical problems in processing plasma for MP measurement, the use of platelet-free plasma may be preferred. When frozen-thawed plasma is used, especially PMP and annexinV-positive MP counts should be interpreted with caution. When flow cytometry is chosen as a method for quantifi-cation of MPs, some analytical conditions should be standardised, e.g. settings of the flow cytometer, quality of the antibodies, and use of counting beads. Fluorescence-nanoparticle tracking analysis and atomic force microscopy can accurately count nanosized MPs, but unfortunately the operational procedures of both methods are still time consuming and they give no information on the functional properties of MPs. The MP-TF activity assay provides information on MPs carrying active TF, regardless of their parental origin. Ultimately, standardisation of pre-analytical procedures and the introduction of reliable and rapid methods for the measurement of MPs are urgently needed to facilitate their use as biomarker in the pathophysiology of diseases.

Keywords

Microparticles - tissue factor /factor VII - annexins - blood products - proteomics

• References

• 1 Wolf P. The nature and significance of platelet products in human plasma. Br J Haematol 1967; 13: 269-288.
• 2 Zwaal RF, Comfurius P, Bevers EM. Surface exposure of phosphatidylserine in pathological cells. Cell Mol Life Sci 2005; 62: 971-988.
• 3 Nieuwland R, Sturk A. Why do cells release vesicles?. Thromb Res 2010; 125 (Suppl. 01) S49-S51.
• 4 Burnier L, Fontana P, Kwak BR, Angelillo-Scherrer A. Cell-derived microparticles in haemostasis and vascular medicine. Thromb Haemost 2009; 101: 439-451.
• 5 Cocucci E, Racchetti G, Meldolesi J. Shedding microvesicles: artefacts no more. Trends Cell Biol 2009; 19: 43-51.
• 6 Toth B, Lok CA, Boing A, Diamant M, van der Post JA, Friese K. et al. Microparticles and exosomes: impact on normal and complicated pregnancy. Am J Reprod Immunol 2007; 58: 389-402.
• 7 Thery C, Ostrowski M, Segura E. Membrane vesicles as conveyors of immune responses. Nat Rev Immunol 2009; 9: 581-593.
• 8 Jy W, Horstman LL, Jimenez JJ, Ahn YS, Biro E, Nieuwland R. et al. Measuring circulating cell-derived microparticles. J Thromb Haemost 2004; 2: 1842-1843.
• 9 Castellana D, Kunzelmann C, Freyssinet JM. Pathophysiologic significance of procoagulant microvesicles in cancer disease and progression. Hamostaseologie 2009; 29: 51-57.
• 10 Piccin A, Murphy WG, Smith OP. Circulating microparticles: pathophysiology and clinical implications. Blood Rev 2007; 21: 157-171.
• 11 Martinez MC, Kunzelmann C, Freyssinet JM. Phosphatidylserine and signal transduction: who needs whom?. Sci STKE 2006; 2006: e3.
• 12 Redman CW, Sargent IL. Circulating microparticles in normal pregnancy and pre-eclampsia. Placenta 2008; 29 Suppl A S73-S77.
• 13 Nomura S, Ozaki Y, Ikeda Y. Function and role of microparticles in various clinical settings. Thromb Res 2008; 123: 8-23.
• 14 George FD. Microparticles in vascular diseases. Thromb Res 2008; 122: S55-S59.
• 15 Pap E, Pallinger E, Pasztoi M. et al. Highlights of a new type of intercellular communication: microvesicle-based information transfer. Inflamm Res 2009; 58: 1-8.
• 16 Azevedo LC, Pedro MA, Laurindo FR. Circulating microparticles as therapeutic targets in cardiovascular diseases. Recent Patents Cardiovasc Drug Discov 2007; 2: 41-51.
• 17 Ahn YS. Cell-derived microparticles: ‘Miniature envoys with many faces‘. J Thromb Haemost 2005; 3: 884-887.
• 18 Polack B, Schved JF, Boneu B. Preanalytical recommendations of the ‘Groupe d‘Etude sur l‘Hemostase et la Thrombose’ (GEHT) for venous blood testing in hemostasis laboratories. Haemostasis 2001; 31: 61-68.
• 19 Lippi G, Salvagno GL, Montagnana M, Franchini M, Guidi GC. Venous stasis and routine hematologic testing. Clin Lab Haematol 2006; 28: 332-337.
• 20 Lippi G, Franchini M, Montagnana M. et al. Quality and reliability of routine coagulation testing: can we trust that sample?. Blood Coagul Fibrinolysis 2006; 17: 513-519.
• 21 van Beers EJ, Schaap MC, Berckmans RJ. et al. Circulating erythrocyte-derived microparticles are associated with coagulation activation in sickle cell disease. Haematologica 2009; 94: 1513-1519.
• 22 Hefler L, Grimm C, Leodolter S. et al. To butterfly or to needle: the pilot phase. Ann Intern Med 2004; 140: 935-936.
• 23 van Ierssel SH, Van Craenenbroeck EM, Conraads VM. et al. Flow cytometric detection of endothelial microparticles (EMP): effects of centrifugation and storage alter with the phenotype studied. Thromb Res 2010; 125: 332-339.
• 24 Woywodt A, Blann AD, Kirsch T. et al. Isolation and enumeration of circulating endothelial cells by immunomagnetic isolation: proposal of a definition and a consensus protocol. J Thromb Haemost 2006; 4: 671-677.
• 25 Ahnadi CE, Sabrinah CE, Lepine M. et al. Assessment of platelet activation in several different anticoagulants by the Advia 120 Hematology System, fluorescence flow cytometry, and electron microscopy. Thromb Haemost 2003; 90: 940-948.
• 26 Macey M, Azam U, McCarthy D. et al. Evaluation of the anticoagulants EDTA and citrate, theophylline, adenosine, and dipyridamole (CTAD) for assessing platelet activation on the ADVIA 120 hematology system. Clin Chem 2002; 48: 891-899.
• 27 Neufeld M, Nowak-Gottl U, Junker R. Citrate-theophylline-adenine-dipyridamol buffer is preferable to citrate buffer as an anticoagulant for flow cytometric measurement of platelet activation. Clin Chem 1999; 45: 2030-2033.
• 28 Piersma SR, Broxterman HJ, Kapci M. et al. Proteomics of the TRAP-induced platelet releasate. J Proteomics 2009; 72: 91-109.
• 29 Smalley DM, Root KE, Cho H. et al. Proteomic discovery of 21 proteins expressed in human plasma-derived but not platelet-derived microparticles. Thromb Haemost 2007; 97: 67-80.
• 30 Garcia BA, Smalley DM, Cho H. et al. The platelet microparticle proteome. J Proteome Res 2005; 4: 1516-1521.
• 31 Ueba T, Haze T, Sugiyama M. et al. Level, distribution and correlates of platelet-derived microparticles in healthy individuals with special reference to the metabolic syndrome. Thromb Haemost 2008; 100: 280-285.
• 32 Nomura S, Shouzu A, Taomoto K. et al. Assessment of an ELISA kit for platelet-derived microparticles by joint research at many institutes in Japan. J Atheroscler Thromb 2009; 16: 878-887.
• 33 Hron G, Kollars M, Weber H. et al. Tissue factor-positive microparticles: Cellular origin and association with coagulation activation in patients with colorectal cancer. Thromb Haemost 2007; 97: 119-123.
• 34 Aras O, Shet A, Bach RR. et al. Induction of microparticle- and cell-associated intravascular tissue factor in human endotoxemia. Blood 2004; 103: 4545-4553.
• 35 Breimo ES, Osterud B. Generation of tissue factor-rich microparticles in an ex vivo whole blood model. Blood Coagul Fibrinolysis 2005; 16: 399-405.
• 36 Brunialti MK, Kallas EG, Freudenberg M. et al. Influence of EDTA and heparin on lipopolysaccharide binding and cell activation, evaluated at single-cell level in whole blood. Cytometry 2002; 50: 14-18.
• 37 Shah MD, Bergeron AL, Dong JF. et al. Flow cytometric measurement of micro-particles: pitfalls and protocol modifications. Platelets 2008; 19: 365-372.
• 38 Connor DE, Exner T, Ma DD. et al. Detection of the procoagulant activity of microparticle-associated phosphatidylserine using XACT. Blood Coagul Fibrinolysis 2009; 20: 558-564.
• 39 Contant G, Gouault-Heilmann M, Martinoli JL. Heparin inactivation during blood storage: its prevention by blood collection in citric acid, theophylline, adenosine, dipyridamole-C.T.A.D. mixture. Thromb Res 1983; 31: 365-374.
• 40 Kim HK, Song KS, Lee ES. et al. Optimized flow cytometric assay for the measurement of platelet microparticles in plasma: pre-analytic and analytic considerations. Blood Coagul Fibrinolysis 2002; 13: 393-397.
• 41 CLSI.. Collection, Transport, and Processing of Blood Specimens for Testing Plasma-Based Coagulation Assays and Molecular Hemostasis Assays. Approved Guideline-Fifth Edition. CLSI document H21-A5. 2008. Wayne, PA: Clinical and Laboratory Standards Institute;
• 42 Horstman LL, Jy W, Jimenez JJ. et al. New horizons in the analysis of circulating cell-derived microparticles. Keio J Med 2004; 53: 210-230.
• 43 Weber H, Kollars M, Kyrle PA. et al. Comparison of different methods for isolation and storage of microparticles from human blood. J Thromb Haemost 2007; 5 (Suppl. 02) P-M-445.
• 44 Favaloro EJ, Soltani S, McDonald J. Potential laboratory misdiagnosis of hemophilia and von Willebrand disorder owing to cold activation of blood samples for testing. Am J Clin Pathol 2004; 122: 686-692.
• 45 Bohm M, Taschner S, Kretzschmar E. et al. Cold storage of citrated whole blood induces drastic time-dependent losses in factor VIII and von Willebrand factor: potential for misdiagnosis of haemophilia and von Willebrand disease. Blood Coagul Fibrinolysis 2006; 17: 39-45.
• 46 van Geest-Daalderop JH, Mulder AB, Boonman-de Winter LJ. et al. Preanalytical variables and off-site blood collection: influences on the results of the prothrombin time/international normalized ratio test and implications for monitoring of oral anticoagulant therapy. Clin Chem 2005; 51: 561-568.
• 47 Gelderman MP, Simak J. Flow cytometric analysis of cell membrane microparticles. Methods Mol Biol 2008; 484: 79-93.
• 48 Trummer A, De Rop C, Tiede A. et al. Recovery and composition of microparticles after snap-freezing depends on thawing temperature. Blood Coagul Fibrinol 2009; 20: 52-56.
• 49 Tesselaar ME, Romijn FP, van der Linden IK. et al. Microparticle-associated tissue factor activity: a link between cancer and thrombosis?. J Thromb Haemost 2007; 5: 520-527.
• 50 van der Zee PM, Biro E, Ko Y. et al. P-selectin- and CD63-exposing platelet micro-particles reflect platelet activation in peripheral arterial disease and myocardial infarction. Clin Chem 2006; 52: 657-664.
• 51 Robert S, Poncelet P, Lacroix R. et al. Standardization of platelet-derived micro-particle counting using calibrated beads and a Cytomics FC500 routine flow cyto-meter: a first step towards multicenter studies?. J Thromb Haemost 2009; 7: 190-197.
• 52 Khorana AA, Francis CW, Menzies KE. et al. Plasma tissue factor may be predictive of venous thromboembolism in pancreatic cancer. J Thromb Haemost 2008; 6: 1983-1985.
• 53 Ay C, Freyssinet JM, Sailer T. et al. Circulating procoagulant microparticles in patients with venous thromboembolism. Thromb Res 2009; 123: 724-726.
• 54 Mobarrez F, Antovic J, Egberg N. et al. A multicolor flow cytometric assay for measurement of platelet-derived microparticles. Thromb Res 2010; 125: e110-e116.
• 55 Furie B, Furie BC. Cancer-associated thrombosis. Blood Cells Mol Dis 2006; 36: 177-181.
• 56 Bal L, Ederhy S, Di AE. et al. Circulating procoagulant microparticles in acute pulmonary embolism: A case-control study. Int J Cardiol 2009; 2010; 145: 321-322.
• 57 Steppich BA, Braun SL, Stein A. et al. Plasma TF activity predicts cardiovascular mortality in patients with acute myocardial infarction. Thromb J 2009; 7: 11.
• 58 Yuana Y, Oosterkamp TH, Bahatyrova S. et al. Atomic force microscopy: a novel approach to the detection of nanosized blood microparticles. J Thromb Haemost 2010; 8: 315-323.
• 59 Miguet L, Pacaud K, Felden C. et al. Proteomic analysis of malignant lymphocyte membrane microparticles using double ionization coverage optimization. Proteomics 2006; 6: 153-171.
• 60 Van Der Pol E, Hoekstra AG, Sturk A. et al. Optical and non-optical methods for detection and characterisation of microparticles and exosomes. J Thromb Haemost 2010; 8: 2596-2607.
• 61 Shet AS, Aras O, Gupta K. et al. Sickle blood contains tissue factor-positive micro-particles derived from endothelial cells and monocytes. Blood 2003; 102: 2678-2683.
• 62 Lacroix R, Robert S, Poncelet P. et al. on behalf of the ISTH SSC Workshop.. Standardization of platelet-derived microparticle enumeration by flow cytometry using calibrated beads: results of ISTH SSC collaborative workshop. J Thromb Haemost 2010; 8: 2571-2574.
• 63 Carter NP, Ormerod MG. Introduction to the principles of flow cytometry. In: Flow cytometry: a pratical approach. 3rd ed. Oxford University Press; 2000. pp. 1-22.
• 64 Connor DE, Exner T, Ma DD. et al. The majority of circulating platelet-derived microparticles fail to bind annexin V, lack phospholipid-dependent procoagulant activity and demonstrate greater expression of glycoprotein Ib. Thromb Haemost 2010; 103: 1044-1052.
• 65 Bernimoulin M, Waters EK, Foy M. et al. Differential stimulation of monocytic cells results in distinct populations of microparticles. J Thromb Haemost 2009; 7: 1019-1028.
• 66 Perez-Pujol S, Marker PH, Key NS. Platelet microparticles are heterogeneous and highly dependent on the activation mechanism: studies using a new digital flow cytometer. Cytometry A 2007; 71: 38-45.
• 67 Jimenez JJ, Jy W, Mauro LM. et al. Endothelial cells release phenotypically and quantitatively distinct microparticles in activation and apoptosis. Thromb Res 2003; 109: 175-180.
• 68 Tait JF, Gibson D. Phospholipid binding of annexin V: effects of calcium and membrane phosphatidylserine content. Arch Biochem Biophys 1992; 298: 187-191.
• 69 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.
• 70 Enjeti AK, Lincz L, Seldon M. Bio-maleimide as a generic stain for detection and quantitation of microparticles. Int J Lab Hematol 2008; 30: 196-199.
• 71 Bratosin D, Mitrofan L, Palii C. et al. Novel fluorescence assay using calcein-AM for the determination of human erythrocyte viability and aging. Cytometry A 2005; 66: 78-84.
• 72 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.
• 73 Enjeti AK, Lincz LF, Seldon M. Detection and measurement of microparticles: an evolving research tool for vascular biology. Semin Thromb Hemost 2007; 33: 771-779.
• 74 Horstman LL, Jy W, Minagar A. et al. Cell-derived microparticles and exosomes in neuroinflammatory disorders. Int Rev Neurobiol 2007; 79: 227-268.
• 75 Zwicker JI, Liebman HA, Neuberg D. et al. Tumor-derived tissue factor-bearing microparticles are associated with venous thromboembolic events in malignancy. Clin Cancer Res 2009; 15: 6830-6840.
• 76 Andreola G, Rivoltini L, Castelli C. et al. Induction of lymphocyte apoptosis by tumor cell secretion of FasL-bearing microvesicles. J Exp Med 2002; 195: 1303-1316.
• 77 Al Nedawi K, Meehan B, Micallef J. et al. Intercellular transfer of the oncogenic receptor EGFRvIII by microvesicles derived from tumour cells. Nat Cell Biol 2008; 10: 619-624.
• 78 Kalinkovich A, Tavor S, Avigdor A. et al. Functional CXCR4-expressing microparticles and SDF-1 correlate with circulating acute myelogenous leukemia cells. Cancer Res 2006; 66: 11013-11020.
• 79 Heijnen HF, Schiel AE, Fijnheer R. et al. Activated platelets release two types of membrane vesicles: microvesicles by surface shedding and exosomes derived from exocytosis of multivesicular bodies and alpha-granules. Blood 1999; 94: 3791-3799.
• 80 Jayachandran M, Litwiller RD, Owen WG. et al. Characterization of blood borne microparticles as markers of premature coronary calcification in newly menopausal women. Am J Physiol Heart Circ Physiol 2008; 295: H931-H938.
• 81 Harris JR. Negative Staining of Thinly Spread Biological Samples. In: Methods in Molecular Biology. Humana Press Inc.; 2007. pp. 107-142.
• 82 Dale GL, Remenyi G, Friese P. Quantitation of microparticles released from coated-platelets. J Thromb Haemost 2005; 3: 2081-2088.
• 83 Habib A, Kunzelmann C, Shamseddeen W. et al. Elevated levels of circulating procoagulant microparticles in patients with beta-thalassemia intermedia. Haematologica 2008; 93: 941-942.
• 84 Aupeix K, Hugel B, Martin T. et al. The significance of shed membrane particles during programmed cell death in vitro, and in vivo, in HIV-1 infection. J Clin Invest 1997; 99: 1546-1554.
• 85 Tesselaar ME, Romijn FP, van der Linden IK. et al. Microparticle-associated tissue factor activity in cancer patients with and without thrombosis. J Thromb Haemost 2009; 7: 1421-1423.
• 86 Tilley RE, Holscher T, Belani R. et al. Tissue factor activity is increased in a combined platelet and microparticle sample from cancer patients. Thromb Res 2008; 122: 604-609.
• 87 Manly DA, Wang J, Glover SL. et al. Increased microparticle tissue factor activity in cancer patients with Venous Thromboembolism. Thromb Res 2010; 125: 511-512.
• 88 Garcia RP, Eikenboom HC, Tesselaar ME. et al. Plasma levels of microparticle-associated tissue factor activity in patients with clinically suspected pulmonary embolism. Thromb Res 2010; 126: 345-349.
• 89 Funderburg NT, Mayne E, Sieg SF. et al. Increased tissue factor expression on circulating monocytes in chronic HIV infection: relationship to in vivo coagulation and immune activation. Blood 2010; 115: 161-167.
• 90 Bogdanov VY, Cimmino G, Tardos JG. et al. Assessment of plasma tissue factor activity in patients presenting with coronary artery disease: limitations of a commercial assay. J Thromb Haemost 2009; 7: 894-897.
• 91 Miguet L, Sanglier S, Schaeffer C. et al. Microparticles: a new tool for plasma membrane sub-cellular proteomic. Subcell Biochem 2007; 43: 21-34.
• 92 Josic D, Clifton JG. Mammalian plasma membrane proteomics. Proteomics 2007; 7: 3010-3029.
• 93 Wu CC, MacCoss MJ, Howell KE. et al. A method for the comprehensive proteomic analysis of membrane proteins. Nat Biotechnol 2003; 21: 532-538.
• 94 Smalley DM, Ley K. Plasma-derived microparticles for biomarker discovery. Clin Lab 2008; 54: 67-79.
• 95 Cho J, Furie BC, Coughlin SR. et al. A critical role for extracellular protein disulfide isomerase during thrombus formation in mice. J Clin Invest 2008; 118: 1123-1131.
• 96 Reinhardt C, von Bruhl ML, Manukyan D. et al. Protein disulfide isomerase acts as an injury response signal that enhances fibrin generation via tissue factor activation. J Clin Invest 2008; 118: 1110-1122.
• 97 Versteeg HH, Ruf W. Tissue factor coagulant function is enhanced by protein-disulfide isomerase independent of oxidoreductase activity. J Biol Chem 2007; 282: 25416-25424.
• 98 Raturi A, Miersch S, Hudson JW. et al. Platelet microparticle-associated protein disulfide isomerase promotes platelet aggregation and inactivates insulin. Biochim Biophys Acta 2008; 1778: 2790-2796.
• 99 Hoffman RA, Johnson TS, Britt WB. Flow cytometric electronic direct current volume and radiofrequency impedance measurements of single cells and particles. Cytometry 1981; 1: 377-384.
• 100 Zwicker JI. Predictive value of tissue factor bearing microparticles in cancer associated thrombosis. Thromb Res 2010; 125 (Suppl. 02) S89-S91.
• 101 Kaszuba M, McKnight D, Connah MT. et al. Measuring sub nanometre sizes using dynamic light scattering. J Nanoparticle Res 2008; 10: 823-829.
• 102 Lawrie AS, Albanyan A, Cardigan RA. et al. Microparticle sizing by dynamic light scattering in fresh-frozen plasma. Vox Sang 2009; 96: 206-212.
• 103 Harrison P, Dragovic R, Albanyan A. et al. Application of dynamic light scattering to the measurement of microparticles. J Thromb Haemost 2009; 7 (Suppl. 02) OC-TU-056.
• 104 Filipe V, Hawe A, Jiskoot W. Critical evaluation of Nanoparticle Tracking Analysis (NTA) by NanoSight for the measurement of nanoparticles and protein aggregates. Pharm Res 2010; 27: 796-810.
• 105 Parot P, Dufrene YF, Hinterdorfer P. et al. Past, present and future of atomic force microscopy in life sciences and medicine. J Mol Recognit 2007; 20: 418-431.
• 106 Ricci D, Braga PC. How the Atomic Force Microscopy Works. In: Methods in Molecular Biology: Biomedical Methods and Applications. Humana Press Inc; 2004. pp. 3-12.
• 107 Muller DJ, Amrein M, Engel A. Adsorption of biological molecules to a solid support for scanning probe microscopy. J Struct Biol 1997; 119: 172-188.
• 108 Siedlecki CA, Wang IW, Higashi JM. et al. Platelet-derived microparticles on synthetic surfaces observed by atomic force microscopy and fluorescence microscopy. Biomaterials 1999; 20: 1521-1529.
• 109 Baran J, Baj-Krzyworzeka M, Weglarczyk K. et al. Circulating tumour-derived microvesicles in plasma of gastric cancer patients. Cancer Immunol Immunother 2010; 59: 841-850.
• 110 Berckmans RJ, Neiuwland R, Boing AN. et al. Cell-derived microparticles circulate in healthy humans and support low grade thrombin generation. Thromb Haemost 2001; 85: 639-646.
• 111 Sabatier F, Darmon P, Hugel B. et al. Type 1 and type 2 diabetic patients display different patterns of cellular microparticles. Diabetes 2002; 51: 2840-2845.
• 112 Enjeti AK, Lincz LF, Scorgie FE. et al. Circulating microparticles are elevated in carriers of Factor V Leiden. Thromb Res 2010; 126: 250-253.
• 113 Pereira J, Alfaro G, Goycoolea M. et al. Circulating platelet-derived microparticles in systemic lupus erythematosus. Association with increased thrombin generation and procoagulant state. Thromb Haemost 2006; 95: 94-99.