背景:输血相关急性肺损伤(TRALI)仍然是输血相关死亡率的主要因素。虽然TRALI的发病机制尚不清楚,有证据表明血液成分有作用。因此,我们研究了新鲜冷冻血浆(FFP)的潜在影响,冷沉淀,和来自这些血液成分的细胞外囊泡(EV),对人肺微血管内皮细胞(HLMVECs)体外活力的影响。
方法:使用尺寸排阻色谱法从FFP和冷沉淀中分离出EV,并通过纳米粒子跟踪分析进行表征,西方印迹,和透射电子显微镜。在存在和不存在嗜中性粒细胞的情况下,检查了这些血液成分及其EV对HLMVEC活力的潜在影响(通过锥虫蓝排除法确定),无论是否事先用LPS治疗HLMVECs。
结果:从FFP和冷沉淀中分离出的EV显示出符合最新国际标准的形态和生化特性。而FFP,冷沉淀,和衍生自FFP的电动汽车,每个人都降低了HLMVEC的生存能力,对于来自冷沉淀的EV没有观察到影响。
结论:我们的研究结果表明,FFP的作用存在明显差异,冷沉淀,以及它们各自的EV对HLMVEC的体外生存力。审查这些差异背后的机制可能会导致对促进TRALI发展的因素的更好理解。
BACKGROUND: Transfusion-related acute lung injury (TRALI) remains a major contributor to transfusion-associated mortality. While the pathogenesis of TRALI remains unclear, there is evidence of a role for blood components. We therefore investigated the potential effects of fresh frozen plasma (
FFP), cryoprecipitate, and extracellular vesicles (EVs) derived from these blood components, on the viability of human lung microvascular endothelial cells (HLMVECs) in vitro.
METHODS: EVs were isolated from
FFP and cryoprecipitate using size-exclusion chromatography and characterized by nanoparticle tracking analysis, western blotting, and transmission electron microscopy. The potential effects of these blood components and their EVs on HLMVEC viability (determined by trypan blue exclusion) were examined in the presence and absence of neutrophils, either with or without prior treatment of HLMVECs with LPS.
RESULTS: EVs isolated from
FFP and cryoprecipitate displayed morphological and biochemical properties conforming to latest international criteria. While
FFP, cryoprecipitate, and EVs derived from
FFP, each reduced HLMVEC viability, no effect was observed for EVs derived from cryoprecipitate.
CONCLUSIONS: Our findings demonstrate clear differences in the effects of
FFP, cryoprecipitate, and their respective EVs on HLMVEC viability in vitro. Examination of the mechanisms underlying these differences may lead to an improved understanding of the factors that promote development of TRALI.