Flow cytometric analysis of microparticle phenotype and their role in thrombin generation

Flow cytometric analysis of microparticle phenotype and their role in thrombin generation

Background: Microparticles may be generated from a number of cell types and are known to play a role in haemostasis by a variety of mechanisms. We investigated the role of platelet, red cell, and leucocyte‐derived microparticles in the measurement of thrombin generation. Methods: Four parameters of...

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Published in:Cytometry. Part B, Clinical cytometry Vol. 80B; no. 1; pp. 57 - 63
Main Authors: Macey, M. G., Enniks, N., Bevan, S.
Format: Journal Article
Language:English
Published: Hoboken Wiley Subscription Services, Inc., A Wiley Company 01-01-2011
Subjects:
Adult
calibrated automated thrombogram® (CAT) thrombin generation assay
CD15
CD45
Cell-Derived Microparticles - genetics
Cell-Derived Microparticles - physiology
Female
Flow Cytometry
Glycophorin - analysis
Humans
Immunophenotyping
Leukocyte Common Antigens - blood
Lewis X Antigen - blood
Male
Middle Aged
Phenotype
plasma filtration
Platelet Glycoprotein GPIb-IX Complex - analysis
Thrombin - biosynthesis
Thrombin - genetics
Young Adult
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Summary:Background: Microparticles may be generated from a number of cell types and are known to play a role in haemostasis by a variety of mechanisms. We investigated the role of platelet, red cell, and leucocyte‐derived microparticles in the measurement of thrombin generation. Methods: Four parameters of thrombin generation (the endogenous thrombin potential (ETP), lag time, time to peak, peak height) and microparticle content was determined in 35 plasma samples from normal individuals pre and post filtration to remove microparticles. Immunofluorescent flow cytometry was used to identify and enumerate platelet, leucocyte, monocyte and red cell derived microparticles in plasma samples based on the expression of CD42b, CD45, CD15, and Glycophorin A respectively. Expression of phosphatidylserine and tissue factor by microparticles was determined by Annexin V and anti CD142 binding. The pre and post filtration results were compared. Results: There was a significant decrease in ETP and Peak Height, and an increase in the time to peak post filtration (P < 0.001). A significant decrease in the number of CD42+, CD45+, CD15+, CD142+, and Annexin V+ microparticles was also observed. The change in CD42b+ microparticles correlated highly with the change in Annexin V+ microparticles (r = 0.68). Whilst the change in ETP correlated best with the change in CD15+ microparticles (r = 0.45) and the change in time to peak correlated with the change in Annexin V binding (r = 0.52) (P < 0.01). Conclusion: The presence of micropartcles in plasma significantly affects thrombin generation. © 2010 International Clinical Cytometry Society
Bibliography:How to cite this article: Macey MG, Enniks N, Bevan S. Flow cytometric analysis of microparticle phenotype and their role in thrombin generation. Cytometry Part B 2011; 80B: 57–63.
ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
ISSN:1552-4949
1552-4957
DOI:10.1002/cyto.b.20551