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The basis examination of leukocyte–platelet aggregates with CD 45 gating as a novel platelet activation marker
Author(s) -
Nagasawa A.,
Matsuno K.,
Tamura S.,
Hayasaka K.,
Shimizu C.,
Moriyama T.
Publication year - 2013
Publication title -
international journal of laboratory hematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.705
H-Index - 55
eISSN - 1751-553X
pISSN - 1751-5521
DOI - 10.1111/ijlh.12051
Subject(s) - platelet , platelet activation , chemistry , monocyte , flow cytometry , microbiology and biotechnology , monoclonal antibody , mean platelet volume , immunology , population , thrombin , antibody , biology , medicine , environmental health
Summary Introduction Platelet activation in circulation is considered to be associated with thrombosis and inflammation; thus, sensitive and easy‐to‐use markers are necessary. In this study, we established a simple and rapid protocol to clinically examine leukocyte–platelet aggregate formation associated with activated platelets in circulation. Methods Whole blood was stained with PC 5‐conjugated anti‐ CD 45 monoclonal antibody and fluorescent isothiocyanate‐conjugated anti‐ CD 41 monoclonal antibody for leukocyte–platelet aggregate analysis. For platelet activation, 5 μ m thrombin receptor–activated peptide ( TRAP ) or 2 μg/mL collagen was added. Samples were analyzed by EPICS XL (Beckman Coulter, Miami, FL, USA). Monocytes, neutrophils, and lymphocytes were gated based on differences in CD 45 fluorescence intensity and side scatter. For each gate, the percentage (%) of platelets expressing CD 41 was analyzed. Same drawing sample was stained with anti‐ CD 62P monoclonal antibody. Platelet CD 62P expression was then analyzed with gating for platelet cell population. Results We analyzed leukocyte–platelet aggregates and platelet CD 62P expression in 18 healthy individuals. Leukocyte–platelet aggregates, mainly monocyte–platelet aggregates, increased when platelets were activated by platelet agonists. Monocyte–platelet aggregates and neutrophil–platelet aggregates also increased over time with mild platelet activation. Conclusion Leukocyte–platelet aggregates, mainly monocyte–platelet aggregates, appear to be a sensitive marker of platelet activation in circulation.