Novel whole blood assay for phenotyping platelet reactivity in mice identifies ICAM-1 as a mediator of platelet-monocyte interaction
Author(s)
Type
Journal Article
Abstract
Testing of platelet function is central to the cardiovascular phenotyping of genetically modified mice. Traditional platelet function tests have been developed primarily for testing human samples and the volumes required make them highly unsuitable for the testing of mouse platelets. This limits research in this area. To address this problem, we have developed a miniaturized whole blood aggregometry assay, based on a readily accessible 96-well plate format coupled with quantification of single platelet depletion by flow cytometric analysis. Using this approach, we observed a concentration-dependent loss of single platelets in blood exposed to arachidonic acid, collagen, U46619 or protease activated receptor 4 activating peptide. This loss was sensitive to well-established antiplatelet agents and genetic manipulation of platelet activation pathways. Observations were more deeply analyzed by flow cytometric imaging, confocal imaging, and measurement of platelet releasates. Phenotypic analysis of the reactivity of platelets taken from mice lacking intercellular adhesion molecule (ICAM)-1 identified a marked decrease in fibrinogen-dependent platelet-monocyte interactions, especially under inflammatory conditions. Such findings exemplify the value of screening platelet phenotypes of genetically modified mice and shed further light upon the roles and interactions of platelets in inflammation.
Date Issued
2015-09-03
Date Acceptance
2015-07-19
Citation
Blood, 2015, 126 (10), pp.e11-e18
ISSN
0006-4971
Publisher
American Society of Hematology
Start Page
e11
End Page
e18
Journal / Book Title
Blood
Volume
126
Issue
10
Copyright Statement
© 2015 The American Society of Hematology
Subjects
Science & Technology
Life Sciences & Biomedicine
Hematology
LEUKOCYTE-ENDOTHELIUM INTERACTION
P2Y(12) RECEPTOR
THROMBOXANE A(2)
FLOW-CYTOMETRY
ADHESION
AGGREGATION
AGGREGOMETRY
ACTIVATION
FIBRINOGEN
INFLAMMATION
Animals
Blood Platelets
Flow Cytometry
Intercellular Adhesion Molecule-1
Mice
Mice, Inbred C57BL
Mice, Knockout
Microscopy, Confocal
Monocytes
Phenotype
Platelet Aggregation
Platelet Function Tests
Sensitivity and Specificity
Blood Platelets
Monocytes
Animals
Mice, Inbred C57BL
Mice, Knockout
Mice
Intercellular Adhesion Molecule-1
Microscopy, Confocal
Platelet Function Tests
Flow Cytometry
Sensitivity and Specificity
Platelet Aggregation
Phenotype
1102 Cardiorespiratory Medicine and Haematology
1103 Clinical Sciences
1114 Paediatrics and Reproductive Medicine
Immunology
Publication Status
Published
Date Publish Online
2015-07-27