The study of Platelet Function using Flow Cytometry to Assess Effectiveness of Platelets Transfusion Therapy

BACKGROUND: Post-transfusion corrected count increments (CCI) following transfusion of platelets have been used as the indicator for assessing effectiveness of platelet transfusion therapy, however, CCI is not a indicator for the status of platelet function in transfused patients. Nevertheless, there are few reports that have studied the platelet function in patients with platelets transfusion. The evaluation of the platelets functions in patients before and after platelets transfusion were investigated to be helpful inducing the effective platelets transfusion. METHOD: Whole blood samples were obtained from 30 healthy donors and 30 cases of 26 patients (AML 20, ALL 1, CML 3, aplastic anemia 2) with platelets transfusion. Diluted blood samples, either in the resting state or in activated state by the addition of the agonist, 20 μM ADP or 100 μM TRAP, were stained with fluorescein conjugated monoclonal antibody of PAC1 or CD62P. Then, the percentages of expression for each marker were analyzed by flow cytometry. For leukocytes-platelet aggregates, monoclonal antibodies of CD41a, CD14 and CD45 were added simultaneously to undiluted whole blood samples for leukocytes-platelet aggregates. The platelet counts for CCI and platelet parameters were performed with ADVIA 120(Bayer, USA). RESULTS: 1) Reference values for the percentages of expression of PAC1 and CD62P in platelets, respectively, were 0.1~12.5% and 0.0~4.7% at the resting status, 65.3~92.4% and 39.0~75.7% at the addition of 20 μM ADP, 68.1~93.1% and 60.5~91.2% at the addition of 100 μM TRAP, and leukocyte-platelet aggregates 2.8~23.6%, granulocyte-platelet aggregates 5.3~34.2%, lymphocyte-platelets aggregates 4.9~21.6%. 2) The range of CCI in patients was -3248~18603 m2/uL, CCI showed significantly correlated with the percentage of expression of CD62P with activated with ADP in patients after transfusion (r=0.384, P<0.05). Although the platelet functions of patients were increased after platelet transfusion, the activity were 20~40% before transfusion, and 40~60% after platelet transfusions compared to control group. 3) There was no significant difference of characteristics or effectiveness of transfused platelets between random platelet concentrates and single donor platelets. CONCLUSIONS: It is anticipated that reference values for the platelet activation markers of platelets in resting status or the activated status with agonists and leukocyte-platelet aggregates should be helpful to interpreting platelet function test by flow cytometry in our laboratory, and the platelet counts in patients with hemato-oncologic disorders must be interpreted with caution, because their platelets functions were depressed as compared to the control group, despite platelets transfusion.

Author dc:creator

• 김, 신원

Contributors dc:contributor

• 임, 영애
• 대학원 의학과
• 199924313

• 혈소판 수혈
• 혈소판 기능
• CCI(corrected count increments)
• 유세포 분석기
• 백혈구-혈소판 응집

Language dc:language

ko

Identifier

http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000001862
000000001862

OAI identifier oai:identifier

oai:repository.ajou.ac.kr:201003/1617