Sut Caroline, Hamzeh-Cognasse Hind, Arthaud Charles-Antoine, Eyraud Marie-Ange, Chettab Kamel, Dumontet Charles, Laradi Sandrine, Burnouf Thierry, Garraud Olivier, Cognasse Fabrice
Université de Lyon, GIMAP-EA3064, Saint-Etienne, France.
Établissement Français du Sang, Auvergne-Rhône-Alpes, Saint-Etienne, France.
Transfusion. 2018 Nov;58(11):2635-2644. doi: 10.1111/trf.14973. Epub 2018 Oct 16.
Platelet transfusions are safe but can nevertheless cause serious adverse reactions (SARs). This study investigated the effects of platelet biological response modifiers (BRMs) that accumulate during storage and are commonly associated with transfusion adverse reactions.
Endothelial cells (ECs), that is, EA.hy926, were exposed in vitro to supernatants of platelet components (PCs) that had been either implicated or not in SARs. The EC Biology RT Profiler PCR Array was used at the same time to study 84 genes related to functions of ECs. Soluble cytokines and surface expression of EC markers were determined by Luminex/enzyme-linked immunosorbent assay technology and flow cytometry, respectively. Apoptosis and scratch wound assays were performed using IncuCyte technology.
In vitro exposure of EA.hy926 monolayers with Day 0, 1-2, and 3-4 stored PC supernatants resulted in decreases in surface expression of markers of ECs. There was differential production of soluble BRMs in the tested cell line. Exposure to the supernatants of PCs that had been implicated in SARs showed a significant difference in the expression of the EC surface markers. EC mediators also responded differently when exposed to PC supernatants of different storage times and PCs involved in SARs.
PC supernatants collected at Day 1-2 activate fewer cell lines of ECs compared with supernatants collected at Day 3-4. Moreover, PC supernatants involved in SARs appear to alter EC activation compared with the control and storage length.
血小板输注是安全的，但仍可能引起严重不良反应（SARs）。本研究调查了在储存过程中积累且通常与输血不良反应相关的血小板生物反应调节剂（BRMs）的作用。
将内皮细胞（ECs），即EA.hy926，在体外暴露于已被证实或未被证实与SARs有关的血小板成分（PCs）的上清液中。同时使用EC生物学RT Profiler PCR阵列研究与EC功能相关的84个基因。可溶性细胞因子和EC标志物的表面表达分别通过Luminex/酶联免疫吸附测定技术和流式细胞术测定。使用IncuCyte技术进行细胞凋亡和划痕试验。
将EA.hy926单层细胞在体外暴露于第0天、1 - 2天和3 - 4天储存的PC上清液中，导致EC标志物的表面表达降低。在测试的细胞系中可溶性BRMs的产生存在差异。暴露于已被证实与SARs有关的PC上清液中，EC表面标志物的表达有显著差异。当暴露于不同储存时间的PC上清液和与SARs有关的PC时，EC介质的反应也不同。
与第3 - 4天收集的上清液相比，第1 - 2天收集的PC上清液激活的EC细胞系较少。此外，与对照和储存时间相比，与SARs有关的PC上清液似乎会改变EC的激活。
