Effects of storage and leukocyte reduction on the concentration and procoagulant activity of extracellular vesicles in canine packed red cells
Effects of storage and leukocyte reduction on the concentration and procoagulant activity of extracellular vesicles in canine packed red cells
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DOI：
10.1111/vec.13050
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发表时间：
2021-03-10
影响因子：
1.3
通讯作者：
Mackman, Nigel
中科院分区：
文献类型：
--
作者：
Avenick, Daniel;Kidd, Linda;Mackman, Nigel
目的描述犬浓缩红细胞（pRBC）中细胞外囊泡（EV）的大小和促凝血活性，以及白细胞减少对这些特征的影响。设计前瞻性队列研究。设置私人小动物专科转诊医院和大学研究实验室。动物10只健康献血犬。干预根据标准方案获得5个pRBC单位，5例在处理前白细胞减少。在第0、10、20、32和42天收集pRBC单位的无血小板上清液。测量和主要结果进行纳米颗粒跟踪分析以确定EV的大小和浓度。使用捕获试验确定与磷脂酰丝氨酸阳性EV相关的凝血酶生成。在通过以20，000 x g离心上清液分离的EV亚组中测量与磷脂酰丝氨酸阳性EV和组织因子阳性EV相关的因子Xa产生。R软件包nparLD和Mann-Whitney U检验分别用于确定储存持续时间的影响和白细胞减少的影响。小（平均< 125 nm）促凝血EV随时间积累，在非白细胞减少和白细胞减少的单位中在第20天或之后发生显著增加。EV的促凝血活性是由于磷脂酰丝氨酸，而不是组织因子。EV浓度和促凝血活性的增加在非白细胞减少单位中发生得更早。细胞外囊泡的积累和促凝血活性没有减少在任何个人的时间点通过白细胞reduction.Conclusions进一步的研究表征和确定的临床相关性的小促凝血EV在pRBC是必要的。
Objectives To characterize the size and procoagulant activity of extracellular vesicles (EV) that accumulate in canine packed red blood cells (pRBCs) over time and the effect of leukocyte reduction on these characteristics.Design Prospective cohort study.Setting Private small animal specialty referral hospital and university research laboratories.Animals Ten healthy blood donor dogs.Interventions Five pRBCs units were obtained according to standard protocols, and 5 were leukocyte-reduced prior to processing. Platelet-free supernatant from the pRBC units was collected on days 0, 10, 20, 32, and 42.Measurements and main results Nanoparticle tracking analysis was performed to determine the size and concentration of EVs. Thrombin generation associated with phosphatidylserine-positive EVs was determined using a capture assay. Factor Xa generation associated with phosphatidylserine-positive EVs and tissue factor-positive EVs was measured in a subset of EVs isolated by centrifugation of the supernatant at 20,000 x g. R package nparLD and the Mann-Whitney U-test were used to determine the effect of duration of storage and the effect of leukocyte reduction, respectively. Small (mean < 125 nm) procoagulant EVs accumulated over time, with significant increases occurring on or after day 20 in both non-leukocyte reduced and leukocyte-reduced units. The procoagulant activity of the EVs was due to phosphatidylserine, not tissue factor. Increases in EV concentration and procoagulant activity occurred earlier in non-leukocyte reduced units. Extracellular vesicle accumulation and procoagulant activity were not decreased at any individual time point by leukocyte reduction.Conclusions Further studies characterizing and determining the clinical relevance of small procoagulant EVs in pRBCs are warranted.