Abstract
Cell-derived microparticles (MPs) are increasingly recognized as important cell-to-cell signaling mechanisms and may exhibit important functions in homeostasis but also in pathogenesis. Indeed, MPs are associated with a number of diseases inhibiting their production that protects against pathogenesis. MPs are distinct from exosomes and apoptotic bodies, often exhibiting the membrane proteins of the activated or apoptotic cell from which they are derived. Electron microscopic analyses have shown that MPs are produced by all cell types tested to date, and ELISA-based assays have established that increased numbers of MPs are produced following cell activation. These approaches do not, however, determine the exact number of MPs and distribution of functional proteins on their surface. Flow cytometry represents an obvious approach to analyze MPs, and we present here a method to assess the number and phenotype of MPs by using a conventional flow cytometer. We also present the caveats with this method and describe a new imaging flow cytometry approach that overcomes these limitations.
An erratum to this chapter can be found at http://dx.doi.org/10.1007/978-1-61737-950-5_24
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Reininger, A. J., Heijnen, H. F., Schumann, H., Specht, H. M., Schramm, W., and Ruggeri, Z. M. (2006) Mechanism of platelet adhesion to von Willebrand factor and microparticle formation under high shear stress. Blood 107, 3537–3545.
Kushak, R. I., Nestoridi, E., Lambert, J., Selig, M. K., Ingelfinger, J. R., and Grabowski, E. F. (2005) Detached endothelial cells and microparticles as sources of tissue factor activity. Thromb Res 116, 409–419.
Aras, O., Shet, A., Bach, R. R., Hysjulien, J. L., Slungaard, A., Hebbel, R. P., Escolar, G., Jilma, B., and Key, N. S. (2004) Induction of microparticle- and cell-associated intravascular tissue factor in human endotoxemia. Blood 103, 4545–4553.
Beyer, C. and Pisetsky, D. S. (2010) The role of microparticles in the pathogenesis of rheumatic diseases. Nat Rev Rheumatol 6, 21–29.
Combes, V., Coltel, N., Faille, D., Wassmer, S. C., and Grau, G. E. (2006) Cerebral malaria: role of microparticles and platelets in alterations of the blood–brain barrier. Int J Parasitol 36, 541–546.
Combes, V., Coltel, N., Alibert, M., van Eck, M., Raymond, C., Juhan-Vague, I., Grau, G.E., and Chimini, G. (2005) ABCA1 gene deletion protects against cerebral malaria: potential pathogenic role of microÂparticles in neuropathology. Am J Pathol 166, 295–302.
Combes, V., Simon, A. C., Grau, G. E., Arnoux, D., Camoin, L., Sabatier, F., Mutin, M., Sanmarco, M., Sampol, J., and Dignat-George, F. (1999) In vitro generation of endothelial microparticles and possible prothrombotic activity in patients with lupus anticoagulant. J Clin Invest 104, 93–102.
Shet, A.S. (2008) Characterizing blood microparticles: technical aspects and challenges. Vasc Health Risk Manag 4, 769–774.
Bebawy, M., Combes, V., Lee, E., Jaiswal, R., Gong, J., Bonhoure, A., and Grau, G. E. (2009) Membrane microparticles mediate transfer of P-glycoprotein to drug sensitive cancer cells. Leukemia 23, 1643–1649.
Couper, K. N., Barnes, T., Hafalla, J. C., Combes, V., Ryffel, B., Secher, T, Grau, G. E., Riley, E. M., and de Souza, J. B. G. (2010) Parasite-derived plasma microparticles contribute significantly to malaria infection-induced inflammation through potent macrophage stimulation. PLoS Pathog 6, e1000744.
Combes, V., Taylor, T. E., Juhan-Vague, I., Mege, J. L., Mwenechanya, J., Tembo, M., Grau, G. E., and Molyneux, M. E. (2004) Circulating endothelial microparticles in malawian children with severe falciparum malaria complicated with coma. JAMA 291, 2542–2544.
Simak, J., Holada, K., Risitano, A. M., Zivny, J. H., Young, N. S., and Vostal, J. G. (2004) Elevated circulating endothelial membrane microparticles in paroxysmal nocturnal haemoglobinuria. Br J Haematol 125, 804–813.
van der Heyde, H. C., Gramaglia, I., Sun, G., and Woods, C. (2005) Platelet depletion by anti-CD41 (αIIb) mAb injection early but not late in the course of disease protects against Plasmodium berghei pathogenesis by altering the levels of pathogenic cytokines. Blood 105, 1956–1963.
Faille, D., Combes, V., Mitchell, A. J., Fontaine, A., Juhan-Vague, I., Alessi, M-C., Chimini, G., Fusai, T., and Grau, G. E. (2009) Platelet microparticles: a new player in malaria parasite cytoadherence to human brain endothelium. FASEB J 23, 3449–3458.
Bernimoulin, M., Waters, E. K., Foy, M., Steele, B. M., Sullivan, M., Falet, H., Walsh, M. T., Barteneva, N., Geng, J. G., Hartwig, J. H., et al. (2009) Differential stimulation of monocytic cells results in distinct populations of microparticles. J Thromb Haemost 7, 1019–1028.
Gelderman, M. P. and Simak, J. (2008) Flow cytometric analysis of cell membrane microparticles. Methods Mol Biol 484, 79–93.
Horstman, L. L., Jy, W., Jimenez, J. J., Bidot, C., and Ahn, Y. S. (2004) New horizons in the analysis of circulating cell-derived microparticles. Keio J Med 53, 210–230.
Satta, N., Toti, F., Feugeas, O., Bohbot, A., Dachary-Prigent, J., Eschwege, V., Hedman, H., and Freyssinet, J. M. (1994) Monocyte vesiculation is a possible mechanism for dissemination of membrane-associated procoagulant activities and adhesion molecules after stimulation by lipopolysaccharide. J Immunol 153, 3245–3255.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
van der Heyde, H.C., Gramaglia, I., Combes, V., George, T.C., Grau, G.E. (2011). Flow Cytometric Analysis of Microparticles. In: Hawley, T., Hawley, R. (eds) Flow Cytometry Protocols. Methods in Molecular Biology, vol 699. Humana Press. https://doi.org/10.1007/978-1-61737-950-5_16
Download citation
DOI: https://doi.org/10.1007/978-1-61737-950-5_16
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61737-949-9
Online ISBN: 978-1-61737-950-5
eBook Packages: Springer Protocols