Black Anne, Pienimaeki-Roemer Annika, Kenyon Oliver, Orsó Evelyn, Schmitz Gerd
Institute for Laboratory Medicine and Transfusion Medicine, University of Regensburg, Regensburg, Germany.
Apogee Flow Systems Ltd., Hertfordshire, UK.
Transfusion. 2015 Sep;55(9):2184-96. doi: 10.1111/trf.13128. Epub 2015 Jun 1.
Platelet-derived extracellular vesicles (PL-EVs) are present in plateletpheresis concentrates (PCs) and may influence the quality of PCs. The aim of the study was to analyze PC-derived PL-EVs and to correlate them with standard quality control (QC) variables of PCs and with donor-specific laboratory variables.
PL-EVs were analyzed by standard as well as advanced high-sensitivity flow cytometry (FCM) and nanoparticle tracking analysis. A hematology analyzer was applied to the determination of platelet (PLT) count and immature PLT fraction (IPF). Functional capacity of PLTs (CD62P in response to thrombin receptor-activating peptide 6 activation) was measured by FCM. All in vitro measurements were carried out on Day 0 and on Day 5. Altogether, a total of 42 PC samples, 15 irradiated on Day 0, were investigated.
Externalization of CD62P, as an indicator of intact PLT function, significantly decreased during in vitro PLT senescence and CD62P expression inversely correlated with increased PL-EV levels. Interestingly, in fresh PCs a significant correlation was found between PL-EVs and different hemapheresis instruments, duration of apheresis, and IPF count in peripheral blood of the donor before apheresis. In senescent PCs, the body mass index of donors inversely correlated with the PL-EV counts.
Loss of PLT function in PCs was associated with increased PL-EV levels. Shedding of PL-EVs depends on shear stress influenced by different hemapheresis settings and diverse preanalytical conditions of donors. PL-EV analysis may stimulate new quality and apheresis strategies for more vital PLTs for transfusion.
血小板衍生的细胞外囊泡(PL-EVs)存在于单采血小板浓缩物(PCs)中,可能会影响PCs的质量。本研究的目的是分析PCs衍生的PL-EVs,并将它们与PCs的标准质量控制(QC)变量以及供体特异性实验室变量相关联。
通过标准以及先进的高灵敏度流式细胞术(FCM)和纳米颗粒跟踪分析对PL-EVs进行分析。应用血液分析仪测定血小板(PLT)计数和未成熟血小板分数(IPF)。通过FCM测量PLTs的功能能力(对凝血酶受体激活肽6激活的反应中的CD62P)。所有体外测量均在第0天和第5天进行。总共研究了42个PC样本,其中15个在第0天进行了辐照。
作为完整PLT功能指标的CD62P外化在体外PLT衰老过程中显著降低,并且CD62P表达与PL-EV水平升高呈负相关。有趣的是,在新鲜PCs中,发现PL-EVs与不同的血液成分分离仪器、单采持续时间以及单采前供体外周血中的IPF计数之间存在显著相关性。在衰老的PCs中,供体的体重指数与PL-EV计数呈负相关。
PCs中PLT功能的丧失与PL-EV水平升高有关。PL-EVs的释放取决于不同血液成分分离设置和供体不同分析前条件所影响的剪切应力。PL-EV分析可能会激发新的质量和血液成分分离策略,以获得更有活力的用于输血的血小板。