Angénieux Catherine, Couvidou Adèle, Brouard Nathalie, Eckly Anita, Dupuis Arnaud, Mangin Pierre H, Maître Blandine
UMR_S1255, INSERM, Strasbourg, France.
Etablissement Français du Sang-Grand Est, Strasbourg, France.
Res Pract Thromb Haemost. 2023 Feb 14;7(1):100006. doi: 10.1016/j.rpth.2022.100006. eCollection 2023 Jan.
The platelet population is heterogeneous, with different subsets that differ on the basis of their function and reactivity. An intrinsic factor participating in this difference of reactivity could be the platelet age. The lack of relevant tools allowing a formal identification of young platelets prevents so far to draw solid conclusions regarding platelet reactivity. We recently reported that human leukocyte antigen-I (HLA-I) molecules are more expressed on human young platelets.
The aim of this study was to assess platelet reactivity according to their age based on HLA-I expression level.
Platelet activation was assessed by flow cytometry (FC) for different platelet subsets based on their HLA-I expression. These populations were further cell sorted and their intrinsic properties were determined by FC and electron microscopy (EM). Statistical analyses were performed with GraphPad Prism 5.02 software using two-way ANOVA followed by a Tukey post hoc test.
HLA-I expression level allowed the identification of 3 platelet subpopulations regarding to their age (HLA low, dim, and high). HLA-I was reliable to guide platelet cell sorting and highlighted the features of young platelets in the HLA-I population. In response to different soluble agonists, HLA-I platelets were the most reactive subset as shown by the level of P-selectin secretion and fibrinogen binding assessed by flow cytometry. Moreover, the highest capacity of HLA-I platelets to simultaneously express annexin-V and von Willebrand factor or activated αIIbβ3 after coactivation with TRAP and CRP indicated that the procoagulant feature of platelets was age-related.
The young HLA-I population is the most reactive and prone to become procoagulant. These results open up new perspectives to investigate deeply the role of young and old platelets.
血小板群体是异质性的,不同亚群在功能和反应性方面存在差异。参与这种反应性差异的一个内在因素可能是血小板年龄。由于缺乏能够正式鉴定年轻血小板的相关工具,迄今为止,关于血小板反应性难以得出确凿结论。我们最近报道,人类白细胞抗原-I(HLA-I)分子在人类年轻血小板上表达更多。
本研究旨在根据HLA-I表达水平评估不同年龄血小板的反应性。
通过流式细胞术(FC)基于HLA-I表达评估不同血小板亚群的血小板活化情况。对这些群体进一步进行细胞分选,并通过FC和电子显微镜(EM)确定其内在特性。使用GraphPad Prism 5.02软件进行统计分析,采用双向方差分析,随后进行Tukey事后检验。
HLA-I表达水平可根据年龄鉴定出3个血小板亚群(HLA低表达、中等表达和高表达)。HLA-I可可靠地指导血小板细胞分选,并突出了HLA-I群体中年轻血小板的特征。流式细胞术评估显示,在不同可溶性激动剂刺激下,HLA-I血小板是反应性最强的亚群,表现为P-选择素分泌水平和纤维蛋白原结合水平。此外,与凝血酶受体激活肽(TRAP)和C反应蛋白(CRP)共激活后,HLA-I血小板同时表达膜联蛋白-V和血管性血友病因子或活化的αIIbβ3的能力最强,表明血小板的促凝特性与年龄有关。
年轻的HLA-I血小板群体反应性最强,且最容易形成促凝状态。这些结果为深入研究年轻和老年血小板的作用开辟了新的视角。