Flow cytometry analysis of platelet P-selectin expression in whole blood - methodological considerations

Ritchie, J. L.; Hann, Alexander; Rea, Irene Maeve

doi:10.1046/j.1365-2257.2000.00339.x

Cited by 54 publications

(48 citation statements)

References 17 publications

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“…39 Other methods such as fluorescence-activated cell sorting analysis of activationdependent surface markers or b-thromboglobulin plasma levels are more commonly used to assess platelet activation. [40][41][42] Since the fraction of the platelet population that contacts the priming region is rather small ($2%), it would be difficult to detect such low numbers of active platelets among the whole platelet population using these methods. Thus, the combination of platelet margination in flow and a downstream capture region allows for the self-selection of activated platelets, making the current assay more sensitive than any of the bulk fluid-based platelet activation assays.…”

Section: Discussionmentioning

confidence: 99%

Functional assay of antiplatelet drugs based on margination of platelets in flowing blood

2016

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A novel functional assay of antiplatelet drug efficacy was designed by utilizing the phenomena of platelet margination in flowing blood and transient platelet contacts with surface-immobilized platelet agonists. Flow margination enhances transient contacts of platelets with the walls of flow chambers covered with surface-immobilized proteins. Depending on the type and the surface density of the immobilized agonists, such transient interactions could "prime" the marginated platelet subpopulation for enhanced activation and adhesion downstream. By creating an upstream surface patch with an immobilized platelet agonist, platelet flow margination was used to test how effective antiplatelet drugs are in suppressing downstream platelet activation and adhesion. The platelet adhesion downstream was measured by a so-called "capture" patch region close to the distal end of the flow chamber. Platelet adhesion downstream was found to be dose-dependent on the upstream surface coverage of the "priming" patch, with immobilized fibrinogen acting as a platelet agonist. Several antiplatelet agents (acetylsalicylic acid, eptifibatide, and tirofiban) were evaluated for their efficacy in attenuating downstream adhesion after upstream platelet priming. The activation of the platelet population was found to be dependent on both the extent of the upstream agonist stimulus and the antiplatelet drug concentration. Such a relationship provides an opportunity to measure the efficacy of specific antiplatelet agents against the type and concentration of upstream platelet agonists.

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Section: Discussionmentioning

confidence: 99%

Functional assay of antiplatelet drugs based on margination of platelets in flowing blood

2016

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“…from neutrophil degradation start to interfere significantly. The conformation and expression of their membrane-bound adhesion molecules and receptors, however, depend very much on the anticoagulant and transport conditions [40,41]. The temperature speeds up this process, however the cooling of the sample does not prevent it either.…”

Section: Pre-analysis and Anticoagulants Edtamentioning

confidence: 99%

Platelet analysis in laboratory hematology

Nebe¹

2015

LaboratoriumsMedizin

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“…18,19 After activation, platelets will release proinflammatory chemokine, cytokine and adhesion molecule appear onto its surface, including CD62P and CD40 ligand (CD40L). Interaction between platelet and neutrophyl triggers recruitment of neutrophyles to the inflammed tissue, activates the neutrophyles and it contribute toward host immunity and thrombosis process.…”

Section: 19mentioning

confidence: 99%

Comparing P-Selectin (CD62P) expression in patients receiving non-leukodepleted vs leukodepleted thrombocyte concentrates

et al. 2017

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Thrombocyte concentrate (TC) transfusion is an important supportive therapy in patients with thrombocytopenia. The risks in platelet transfusions may be related to the content of TC including the contaminant leukocytes. The aim of this study was to assess the risk of increased level of P-Selectin (CD62P) expression of non-leukodepleted TC transfusions. This was a quasi-experimental study. Subjects were children patients aged 1-18 years who received a non-leukodepleted or a leukodepleted TC transfusions. Comparison of the proportion of increased expression of CD62P in both groups expressed as relative risk. The subjects consisted of 51 patients who received non-leukodepleted and 52 patients who received leukodepleted TC transfusions. The risk of increased expression of CD62P in patients receiving non-leukodepleted TC transfusions were 2.38 (95%CI:1.60-3.53) times higher than those who received leukodepleted TC. Non-leukodepleted have significant higher risks of increased CD62P expression than leukodepleted TC transfusions. ABSTRAKTransfusi thrombocyte concentrate (TC) merupakan terapi pendukung yang penting pada pasien yang mengalami trombositopenia. Risiko yang terjadi dapat terkait dengan komponen yang terkandung dalam TC termasuk adanya lekosit kontaminan. Penelitian ini bertujuan untuk menilai risiko berupa kejadian peningkatan ekspresi P-selectin (CD62P) pada transfusi TC non-lekodeplesi. Penelitian ini merupakan penelitian eksperimental semu. Subjek penelitian adalah pasien anak usia 1-18 tahun yang mendapat transfusi TC non-lekodeplesi dan lekodeplesi. Perbandingan proporsi peningkatan ekspresi CD62P pada kedua kelompok dinyatakan dalam risiko relatif. Subjek terdiri dari 51 pasien yang mendapatkan transfusi TC non-lekodeplesi dan 52 pasien yang mendapatkan transfusi TC lekodeplesi. Risiko peningkatan ekspresi CD62P pada pasien yang mendapatkan transfusi TC non-lekodeplesi adalah 2,38 (95%CI:1,60-3,53) kali lebih tinggi dibandingkan yang mendapatkan transfusi TC lekodeplesi. Transfusi TC non-lekodeplesi memiliki risiko yang lebih tinggi bermakna dibandingkan TC lekodeplesi.

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Flow cytometry analysis of platelet P-selectin expression in whole blood - methodological considerations

Cited by 54 publications

References 17 publications

Functional assay of antiplatelet drugs based on margination of platelets in flowing blood

Functional assay of antiplatelet drugs based on margination of platelets in flowing blood

Platelet analysis in laboratory hematology

Comparing P-Selectin (CD62P) expression in patients receiving non-leukodepleted vs leukodepleted thrombocyte concentrates

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