Effects of physiologic agonists on canine whole blood flow cytometry assays of leukocyte-platelet aggregation and platelet activation
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Authors
Tarnow, IngeKristensen, Annemarie T.
Krogh, Anne K.R.
Frelinger, Andrew L. III
Barnard, Marc R.
Michelson, Alan D.
UMass Chan Affiliations
Department of PediatricsDocument Type
Journal ArticlePublication Date
2008-06-01Keywords
Adenosine DiphosphateAnimals
Antibodies, Monoclonal
Antigens, CD14
Blood Platelets
Collagen
Dogs
Epinephrine
Flow Cytometry
Integrin beta3
Leukocytes, Mononuclear
Neutrophils
P-Selectin
Platelet Aggregation
Thrombin
Hematology
Oncology
Pediatrics
Metadata
Show full item recordAbstract
Platelets play a role in both the innate and adaptive immune systems. Methods for detecting activated platelets and leukocyte-platelet aggregates (LPAs) are useful for basic and applied research concerning the role of platelets in inflammation and immune disorders. The aim of the study was to develop flow cytometric assays for detection of platelets binding to monocytes and neutrophils and for activated platelets in canine whole blood and to investigate the effect of physiologic agonists. Citrate anticoagulated whole blood was incubated with monoclonal antibodies against CD14 and CD61 for detection of LPAs, and the effect of various agonists was investigated. For detection of activated platelets, whole blood was incubated with monoclonal antibodies against CD62P and against a receptor-induced binding site on fibrinogen (CAP1) with CD61 as a platelet identifier. Isotype controls were prepared in parallel. The individual physiologic agonists ADP, collagen and epinephrine increased LPAs, CD62P and CAP1 binding only modestly. However, combinations of agonists gave more substantial increases. A dose-response relationship was seen using alpha- and gamma-thrombin, and ADP as agonists. In conclusion, we have developed flow cytometry assays to measure LPAs and platelet activation in canine whole blood, and have explored the effect of various physiologic agonists at different concentrations.Source
Vet Immunol Immunopathol. 2008 Jun 15;123(3-4):345-52. Epub 2008 Mar 4. doi 10.1016/j.vetimm.2008.02.016DOI
10.1016/j.vetimm.2008.02.016Permanent Link to this Item
http://hdl.handle.net/20.500.14038/43298PubMed ID
18405981Related Resources
Link to article in PubMedae974a485f413a2113503eed53cd6c53
10.1016/j.vetimm.2008.02.016