Mørk Morten, Nielsen Morten H, Bæk Rikke, Jørgensen Malene M, Pedersen Shona, Kristensen Søren R
Department of Clinical Biochemistry, Aalborg University Hospital, Aalborg, Denmark.
Aalborg AF Study Group, Aalborg University Hospital, Aalborg, Denmark.
TH Open. 2018 Apr 16;2(2):e147-e157. doi: 10.1055/s-0038-1642021. eCollection 2018 Apr.
Tissue factor (TF)-bearing microvesicles (MVs) and exosomes may play a role in hemostasis and thrombosis. MVs may be quantified by flow cytometry (FC)-based detection of phosphatidylserine (PS)-positive submicron particles carrying specific antigens, although interference from lipoproteins complicates this approach. In this study, we evaluated the effect of food intake on blood levels of TF-bearing particles measured by FC and small extracellular vesicles (EVs) measured by a protein microarray-based test termed EV Array. Platelet-free plasma (PFP) was obtained from 20 healthy persons in the fasting state and 75 minutes after consumption of a meal. Postprandial changes in the concentration of PS-positive particles, including subgroups binding labeled antibodies against TF, CD41, CD146, and CD62E, respectively (FC), small EVs (EV Array), and TF antigen and procoagulant phospholipids (PPLs) were measured. Furthermore, we tested the effect on FC results of in vitro addition of lipoproteins to fasting PFP. We found significantly increased plasma concentrations of PS-positive particles and all examined subgroups postprandially, while no changes in small EVs, PPL, or TF antigen levels were found. Levels of all types of particles measured by FC were also elevated by lipoprotein spiking. In conclusion, meal consumption as well as in vitro addition of lipoproteins to fasting plasma induces increased levels of PS-positive particles as measured by FC, including TF-positive subtypes and subtypes exposing other antigens. While the observed postprandial increase may to some extent reflect elevated MV levels, our results indicate a substantial interference from lipoproteins.
携带组织因子（TF）的微泡（MVs）和外泌体可能在止血和血栓形成中发挥作用。MVs可通过基于流式细胞术（FC）检测携带特定抗原的磷脂酰丝氨酸（PS）阳性亚微米颗粒来定量，尽管脂蛋白的干扰使这种方法变得复杂。在本研究中，我们评估了食物摄入对通过FC测量的携带TF颗粒的血液水平以及通过一种基于蛋白质微阵列的检测方法（称为EV Array）测量的小细胞外囊泡（EVs）的影响。从20名处于空腹状态的健康人以及进食后75分钟的健康人获取无血小板血浆（PFP）。测量了餐后PS阳性颗粒浓度的变化，包括分别结合抗TF、CD41、CD146和CD62E标记抗体的亚组（FC）、小EVs（EV Array）以及TF抗原和促凝磷脂（PPLs）。此外，我们测试了在体外向空腹PFP中添加脂蛋白对FC结果的影响。我们发现餐后PS阳性颗粒以及所有检测亚组的血浆浓度显著增加，而小EVs、PPL或TF抗原水平未发现变化。通过FC测量的所有类型颗粒的水平也因脂蛋白加样而升高。总之，进食以及在体外向空腹血浆中添加脂蛋白会导致通过FC测量的PS阳性颗粒水平升高，包括TF阳性亚型和暴露其他抗原的亚型。虽然观察到的餐后增加可能在一定程度上反映了MV水平的升高，但我们的结果表明脂蛋白存在实质性干扰。
