Ethylenediaminetetraacetic acid plus. citrate-theophylfine-adenosine-dipyridamole (EDTA-CTAD): A novel anticoagulant for the flow cytometric assessment of platelet and neutrophil activation ex vivo in whole blood

被引:34
作者
Macey, M [1 ]
McCarthy, D
Azam, U
Milne, T
Golledge, P
Newland, A
机构
[1] Royal London Hosp, Dept Haematol, London E1 1BB, England
[2] Queen Mary Univ London, Sch Biol Sci, London, England
[3] London Chest Hosp, Dept Cardiol, London E2 9JX, England
关键词
ADVIA; 120; angiography; anticoagulant; ethylenediaminetetraacetic acid; citrate-theophylline-adenosine-dipyridamole; citrate; flow cytometry; neutrophil activation; platelet activation;
D O I
10.1002/cyto.b.10001
中图分类号
R446 [实验室诊断]; R-33 [实验医学、医学实验];
学科分类号
1001 ;
摘要
Ethylenediaminetetraacetic acid (EDTA) is the anticoagulant recommended for full blood counts, citrate is recommended for coagulation and platelet studies, and citrate-theophylline-adenosine-dipyridamole (CTAD) inhibits platelet activation. Because the combination of EDTA and CTAD (E/C) is better than EDTA or CTAD alone for measuring platelet parameters on the ADVIA 120 Haematology System, we investigated whether it also offers advantages for the flow cytometric assessment of platelet and/or neutrophil activation and platelet-leucocyte aggregate formation ex vivo. Blood from healthy subjects was collected into E/C or citrate, kept at room temperature or at 4degreesC, and analysed 0 to 360 min later in the ADVIA 120 and by immunofluorescent flow cytometry. Platelet count, mean platelet volume, number of platelet clumps, mean platelet component, numbers of CD62P(+) platelets and platelet-leucocyte aggregates, and expression of GD11b on neutrophils changed little over 360 min in blood with E/C kept at 4degreesC. In contrast, one or more parameter changed when blood was kept with E/C at ambient temperature or with citrate at either temperature. The use of E/C in in vitro and in vivo studies is illustrated. Platelet and neutrophil activation status ex vivo can be reliably assessed if blood is collected into E/C, held at 4degreesC, and analysed within 6 h. (C) 2002 Wiley-Liss, Inc.
引用
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页码:30 / 40
页数:11
相关论文
共 70 条
[21]  
Hagberg IA, 2000, PLATELETS, V11, P137
[22]   A METHOD OF PREPARING BLOOD LEUKOCYTES FOR FLOW-CYTOMETRY WHICH PREVENTS UP-REGULATION OF LEUKOCYTE INTEGRINS [J].
HAMBLIN, A ;
TAYLOR, M ;
BERNHAGEN, J ;
SHAKOOR, Z ;
MAYALL, S ;
NOBLE, G ;
MCCARTHY, D .
JOURNAL OF IMMUNOLOGICAL METHODS, 1992, 146 (02) :219-228
[23]  
HAMBURGER SA, 1990, BLOOD, V75, P550
[24]  
Hartwig JH, 1999, THROMB HAEMOSTASIS, V82, P392
[25]  
HASLETT C, 1985, AM J PATHOL, V119, P101
[26]  
*HEM LAB, 2002, PLAT FLOW CYTOMETRY
[27]   Flow cytometry of platelets for clinical analysis [J].
Hickerson, DHM ;
Bode, AP .
HEMATOLOGY-ONCOLOGY CLINICS OF NORTH AMERICA, 2002, 16 (02) :421-+
[28]   PLATELET VOLUME - LABORATORY MEASUREMENT AND CLINICAL-APPLICATION [J].
JACKSON, SR ;
CARTER, JM .
BLOOD REVIEWS, 1993, 7 (02) :104-113
[29]  
JY W, 1992, J LAB CLIN MED, V119, P334
[30]   Platelet microparticles bind, activate and aggregate neutrophils in vitro [J].
Jy, WC ;
Mao, WW ;
Horstman, LL ;
Tao, JG ;
Ahn, YS .
BLOOD CELLS MOLECULES AND DISEASES, 1995, 21 (22) :217-231