Original Article

Korean J Lab Med 2006; 26(5): 323-328

Published online October 1, 2006

Copyright © Korean Society for Laboratory Medicine.

Establishment of Reference Values for Platelet Activation Markers by Flow Cytometry

Sin Won Kim. M.D. and Young Ae Lim. M.D.

Department of Laboratory Medicine, Ajou University School of Medicine, Suwon, Korea

Correspondence to: 임 영 애
우 442-749 경기도 수원시 영통구 원천동 산 5 아주대학교 의과대학 진단검사의학교실
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*이 논문은 한국학술진흥재단에서 지원한 2005년 지역대학우수과학자지원사업(E00129) 연구비로 수행되었음.

Received: July 13, 2006; Revised: September 4, 2006; Accepted: September 17, 2006


Background : This study was purposed to establish reference values for platelet activation markers and leukocyte-platelet aggregates in the evaluation of the platelet function tests using flow cytometry.
Methods : Whole blood samples were obtained from 30 volunteers of healthy adults. Diluted blood samples, either in the resting state or in activated state by the addition of agonist, 20 μM ADP or 100 μM TRAP, were stained with fluorescent conjugated monoclonal antibody of PAC1 or CD62P. Then, the percentages of expression for each marker were analyzed by flow cytometry. For leukocyte-platelet aggregates, monoclonal antibodies of CD41a, CD14 and CD45 were added simultaneously to undiluted whole blood.
Results : Reference values for the percentages of the expression of PAC1 and CD62P, respectively,were 0.1-12.5% and 0.0-4.7% at the resting state, 65.3-92.4% and 39.0-75.7% with the addition of 20 μM ADP, and 68.1-93.1% and 60.5-91.2% with the addition of 100 μM TRAP. Reference values for leukocyte-platelet aggregates, granulocyte-platelet aggregates, and lymphocyte-platelets aggregates were 2.8-23.6%, 5.3-34.2%, and 4.9-21.6%, respectively.
Conclusions : The platelet activation markers at the resting or an activated state with agonists and leukocyte-platelet aggregates could be analyzed using flow cytometry. These reference values should be helpful in interpreting platelet function tests by flow cytometry.

Keywords: Platelet activation, Reference values, Flow cytometry, Leukocyte-platelet