Flow cytometry for comprehensive assessment of platelet functional activity in response to ADP stimulation

被引：0
作者：
Ponomarenko, Evgeniya A. [1 ,2 ]
Ignatova, Anastasia A. [1 ]
Polokhov, Dmitrii M. [1 ]
Filkova, Aleksandra A. [1 ]
Suntsova, Elena V. [1 ]
Zharkov, Pavel A. [1 ]
Fedorova, Daria V. [1 ]
Pisaryuk, Alexandra S. [3 ,4 ]
Meray, Imad [3 ,4 ]
Kobalava, Zhanna D. [3 ,4 ]
Tukhsanboev, Yokubjon S. [3 ,4 ]
Maschan, Alexey A. [1 ]
Novichkova, Galina A. [1 ]
Sveshnikova, Anastasia N. [1 ,5 ]
Panteleev, Mikhail A. [1 ,2 ,5 ,6 ]
机构：
[1] Russian Minist Healthcare, Natl Med Res Ctr Pediat Hematol Oncol & Immunol, Moscow, Russia
[2] Lomonosov Moscow State Univ, Moscow, Russia
[3] Vinogradov City Clin Hosp, Cardiol Dept, Moscow, Russia
[4] Peoples Friendship Univ Russia RUDN, Inst Med, Moscow, Russia
[5] Russian Acad Sci, Ctr Theoret Problems Physicochem Pharmacol, Moscow, Russia
[6] Russian Minist Healthcare, Natl Med Res Ctr Pediat Hematol Oncol & Immunol, Cellular Hemostasis & Thrombosis Lab, 1 Samory Mashela Str, Moscow 117997, Russia
来源：
EUROPEAN JOURNAL OF HAEMATOLOGY | 2024年 / 112卷 / 04期
关键词：
ADP; flow cytometry; ITP; platelet function; platelet preactivation; P2Y(12) RECEPTOR; INCREASED RISK; WHOLE-BLOOD; ACTIVATION; CLOPIDOGREL; COUNT; ELTROMBOPAG; EVENTS;
D O I：
10.1111/ejh.14144
中图分类号：
R5 [内科学];
学科分类号：
1002 ; 100201 ;
摘要：
ObjectivesFlow cytometry with adenosine diphosphate (ADP) allows to characterize molecular changes of platelet function caused by this physiologically important activation, but the methodology has not been thoroughly investigated, standardized and characterized yet. We analyzed the influence of several major variables and chose optimal conditions for platelet function assessment.MethodsFor activation, 2.5 mu M CaCl2, 5 mu M ADP and antibodies were added to diluted blood and incubated for 15 min. We analyzed kinetics of antibody binding and effects of their addition sequence, agonist concentration, blood dilution, exogenous calcium addition and platelet fixation.ResultsWe tested our protocol on 11 healthy children, 22 healthy adult volunteers, 9 patients after a month on dual antiplatelet therapy after percutaneous coronary intervention (PCI), 7 adult patients and 14 children with immune thrombocytopenia (ITP). We found that our protocol is highly sensitive to ADP stimulation with low percentage of aggregates formation. The assay is also sensitive to platelet function inhibition in post-PCI patients. Finally, platelet preactivation with ITP plasma was stronger and caused increase in activation response to ADP stimulation compared to preactivation with low dose of ADP.ConclusionsOur assay is sensitive to antiplatelet therapy and platelet preactivation in ITP patients under physiological conditions with minimal percentage of aggregates formation.
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收藏
页码：554 / 565
页数：12
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