Measurement of monocyte-platelet aggregates by imaging flow cytometry

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

© 2014 International Society for Advancement of Cytometry. Platelets are subcellular blood elements with a well-established role in haemostasis. Upon activation platelets express P-Selectin (CD62P) on the cell membrane and bind to P-Selectin glycoprotein ligand 1 expressing monocytes, influencing them toward a pro-adhesive and inflammatory phenotype. It is well established that elevated circulating monocyte-platelet aggregates (MPAs) are linked to atherothrombosis in high risk patients. However, whole blood flow cytometry (FCM) has recently shown that circulating MPAs may also occur in the absence of platelet activation, particularly in healthy children. A potential limitation of conventional FCM is the potential for coincident events to resemble monocyte platelet aggregates. Here we report a novel imaging cytometry approach to further characterize monocyte-platelet aggregate formation by P-Selectin dependent and P-Selectin independent mechanisms and distinguish circulating MPAs from coincidental events. Monocytes were identified by expression of the lipopolysachharide receptor (CD14 BV421), while platelets were identified by expression of the glycoprotein Ib (CD42b APC). Differentiation of P-Selectin dependent and P-Selectin independent binding was achieved with AF488 labeled CD62P. Overall analysis of circulating and in vitro generated MPAs by conventional and imaging cytometry methods showed very strong correlation (r2=>0.99, P
Original languageEnglish
Pages (from-to)273-278
JournalCytometry Part A
Volume87
Issue number3
DOIs
Publication statusPublished - 2015

Fingerprint

Monocytes
Flow Cytometry
Blood Platelets
P-Selectin
Platelet Activation
Platelet Glycoprotein GPIb-IX Complex
Hemostasis
Adhesives
Cell Membrane
Phenotype

Cite this

@article{0c8043ed6f2d4d8a89c606e7be9104e2,
title = "Measurement of monocyte-platelet aggregates by imaging flow cytometry",
abstract = "{\circledC} 2014 International Society for Advancement of Cytometry. Platelets are subcellular blood elements with a well-established role in haemostasis. Upon activation platelets express P-Selectin (CD62P) on the cell membrane and bind to P-Selectin glycoprotein ligand 1 expressing monocytes, influencing them toward a pro-adhesive and inflammatory phenotype. It is well established that elevated circulating monocyte-platelet aggregates (MPAs) are linked to atherothrombosis in high risk patients. However, whole blood flow cytometry (FCM) has recently shown that circulating MPAs may also occur in the absence of platelet activation, particularly in healthy children. A potential limitation of conventional FCM is the potential for coincident events to resemble monocyte platelet aggregates. Here we report a novel imaging cytometry approach to further characterize monocyte-platelet aggregate formation by P-Selectin dependent and P-Selectin independent mechanisms and distinguish circulating MPAs from coincidental events. Monocytes were identified by expression of the lipopolysachharide receptor (CD14 BV421), while platelets were identified by expression of the glycoprotein Ib (CD42b APC). Differentiation of P-Selectin dependent and P-Selectin independent binding was achieved with AF488 labeled CD62P. Overall analysis of circulating and in vitro generated MPAs by conventional and imaging cytometry methods showed very strong correlation (r2=>0.99, P",
author = "Henry Hui and Kathryn Fuller and Wendy Erber and Matthew Linden",
year = "2015",
doi = "10.1002/cyto.a.22587",
language = "English",
volume = "87",
pages = "273--278",
journal = "Cytometry, Part A",
issn = "1552-4922",
publisher = "Wiley-Liss",
number = "3",

}

Measurement of monocyte-platelet aggregates by imaging flow cytometry. / Hui, Henry; Fuller, Kathryn; Erber, Wendy; Linden, Matthew.

In: Cytometry Part A, Vol. 87, No. 3, 2015, p. 273-278.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Measurement of monocyte-platelet aggregates by imaging flow cytometry

AU - Hui, Henry

AU - Fuller, Kathryn

AU - Erber, Wendy

AU - Linden, Matthew

PY - 2015

Y1 - 2015

N2 - © 2014 International Society for Advancement of Cytometry. Platelets are subcellular blood elements with a well-established role in haemostasis. Upon activation platelets express P-Selectin (CD62P) on the cell membrane and bind to P-Selectin glycoprotein ligand 1 expressing monocytes, influencing them toward a pro-adhesive and inflammatory phenotype. It is well established that elevated circulating monocyte-platelet aggregates (MPAs) are linked to atherothrombosis in high risk patients. However, whole blood flow cytometry (FCM) has recently shown that circulating MPAs may also occur in the absence of platelet activation, particularly in healthy children. A potential limitation of conventional FCM is the potential for coincident events to resemble monocyte platelet aggregates. Here we report a novel imaging cytometry approach to further characterize monocyte-platelet aggregate formation by P-Selectin dependent and P-Selectin independent mechanisms and distinguish circulating MPAs from coincidental events. Monocytes were identified by expression of the lipopolysachharide receptor (CD14 BV421), while platelets were identified by expression of the glycoprotein Ib (CD42b APC). Differentiation of P-Selectin dependent and P-Selectin independent binding was achieved with AF488 labeled CD62P. Overall analysis of circulating and in vitro generated MPAs by conventional and imaging cytometry methods showed very strong correlation (r2=>0.99, P

AB - © 2014 International Society for Advancement of Cytometry. Platelets are subcellular blood elements with a well-established role in haemostasis. Upon activation platelets express P-Selectin (CD62P) on the cell membrane and bind to P-Selectin glycoprotein ligand 1 expressing monocytes, influencing them toward a pro-adhesive and inflammatory phenotype. It is well established that elevated circulating monocyte-platelet aggregates (MPAs) are linked to atherothrombosis in high risk patients. However, whole blood flow cytometry (FCM) has recently shown that circulating MPAs may also occur in the absence of platelet activation, particularly in healthy children. A potential limitation of conventional FCM is the potential for coincident events to resemble monocyte platelet aggregates. Here we report a novel imaging cytometry approach to further characterize monocyte-platelet aggregate formation by P-Selectin dependent and P-Selectin independent mechanisms and distinguish circulating MPAs from coincidental events. Monocytes were identified by expression of the lipopolysachharide receptor (CD14 BV421), while platelets were identified by expression of the glycoprotein Ib (CD42b APC). Differentiation of P-Selectin dependent and P-Selectin independent binding was achieved with AF488 labeled CD62P. Overall analysis of circulating and in vitro generated MPAs by conventional and imaging cytometry methods showed very strong correlation (r2=>0.99, P

U2 - 10.1002/cyto.a.22587

DO - 10.1002/cyto.a.22587

M3 - Article

VL - 87

SP - 273

EP - 278

JO - Cytometry, Part A

JF - Cytometry, Part A

SN - 1552-4922

IS - 3

ER -