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

Autor: Ponomarenko EA; National Medical Research Center of Pediatric Hematology, Oncology and Immunology named after Dmitry Rogachev, Russian Ministry of Healthcare, Moscow, Russia.; Lomonosov Moscow State University, Moscow, Russia., Ignatova AA; National Medical Research Center of Pediatric Hematology, Oncology and Immunology named after Dmitry Rogachev, Russian Ministry of Healthcare, Moscow, Russia., Polokhov DM; National Medical Research Center of Pediatric Hematology, Oncology and Immunology named after Dmitry Rogachev, Russian Ministry of Healthcare, Moscow, Russia., Filkova AA; National Medical Research Center of Pediatric Hematology, Oncology and Immunology named after Dmitry Rogachev, Russian Ministry of Healthcare, Moscow, Russia., Suntsova EV; National Medical Research Center of Pediatric Hematology, Oncology and Immunology named after Dmitry Rogachev, Russian Ministry of Healthcare, Moscow, Russia., Zharkov PA; National Medical Research Center of Pediatric Hematology, Oncology and Immunology named after Dmitry Rogachev, Russian Ministry of Healthcare, Moscow, Russia., Fedorova DV; National Medical Research Center of Pediatric Hematology, Oncology and Immunology named after Dmitry Rogachev, Russian Ministry of Healthcare, Moscow, Russia., Pisaryuk AS; Cardiology Department, Vinogradov City Clinical Hospital, Moscow, Russia.; Institute of Medicine, Peoples' Friendship University of Russia (RUDN), Moscow, Russia., Meray I; Cardiology Department, Vinogradov City Clinical Hospital, Moscow, Russia.; Institute of Medicine, Peoples' Friendship University of Russia (RUDN), Moscow, Russia., Kobalava ZD; Cardiology Department, Vinogradov City Clinical Hospital, Moscow, Russia.; Institute of Medicine, Peoples' Friendship University of Russia (RUDN), Moscow, Russia., Tukhsanboev YS; Cardiology Department, Vinogradov City Clinical Hospital, Moscow, Russia.; Institute of Medicine, Peoples' Friendship University of Russia (RUDN), Moscow, Russia., Maschan AA; National Medical Research Center of Pediatric Hematology, Oncology and Immunology named after Dmitry Rogachev, Russian Ministry of Healthcare, Moscow, Russia., Novichkova GA; National Medical Research Center of Pediatric Hematology, Oncology and Immunology named after Dmitry Rogachev, Russian Ministry of Healthcare, Moscow, Russia., Sveshnikova AN; National Medical Research Center of Pediatric Hematology, Oncology and Immunology named after Dmitry Rogachev, Russian Ministry of Healthcare, Moscow, Russia.; Center for Theoretical Problems of Physicochemical Pharmacology, Russian Academy of Science, Moscow, Russia., Panteleev MA; National Medical Research Center of Pediatric Hematology, Oncology and Immunology named after Dmitry Rogachev, Russian Ministry of Healthcare, Moscow, Russia.; Lomonosov Moscow State University, Moscow, Russia.; Center for Theoretical Problems of Physicochemical Pharmacology, Russian Academy of Science, Moscow, Russia.
Jazyk: angličtina
Zdroj: European journal of haematology [Eur J Haematol] 2024 Apr; Vol. 112 (4), pp. 554-565. Date of Electronic Publication: 2023 Dec 11.
DOI: 10.1111/ejh.14144
Abstrakt: Objectives: Flow 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.
Methods: For activation, 2.5 μM CaCl 2 , 5 μM 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.
Results: We 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.
Conclusions: Our assay is sensitive to antiplatelet therapy and platelet preactivation in ITP patients under physiological conditions with minimal percentage of aggregates formation.
(© 2023 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)
Databáze: MEDLINE
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