UMR_S1255, INSERM, Strasbourg, France.
Etablissement Français du Sang-Grand Est, Strasbourg, France.
J Thromb Haemost. 2019 Sep;17(9):1511-1521. doi: 10.1111/jth.14537. Epub 2019 Jun 27.
Accurate identification of the proportion of young platelets is important to distinguish peripheral thrombocytopenia from a deficit in platelet production. Young platelets are defined by their higher RNA content and are often assessed as thiazole orange bright (TO ) by flow cytometry. In clinical practice, their proportion is estimated by automatic blood counter according to their greater RNA content, which identifies a so-called immature platelet fraction (IPF). However, the detected IPFs are not strictly identical to the young TO platelet population observed by flow cytometry.
The aim of this study was to assess the reliability of HLA I/major histocompatibility I (MHC I) cell surface expression as a marker of young platelets.
The HLA I/MHC I expression was evaluated by flow cytometry after costaining blood with TO and antibodies directed against HLA I/MHC I molecules.
We found that platelets with a higher expression of plasma membrane-localized MHC I molecules displayed an increased TO staining and a higher content in ribosomal P-antigen. Transfusion experiments in mice showed that the number of MHC I molecules expressed on the cell surface of young murine platelets decreased during platelet aging, reaching basal levels within 24 h. Finally, we demonstrated that for patients with thrombocytopenias, the identification of young platelets is better assessed by the flow cytometric determination of the level of HLA I expression than by TO staining or the use of hematological blood counter.
Overall, our results highlight the relevance of MHC I/HLA I expression as a valuable parameter to identify young platelets.
准确识别年轻血小板的比例对于区分外周血小板减少症和血小板生成不足至关重要。年轻血小板的 RNA 含量较高,通常通过流式细胞术被定义为噻唑橙亮(TO)。在临床实践中,根据其较高的 RNA 含量,自动血液计数器估计其比例,这会识别出所谓的不成熟血小板分数(IPF)。然而,检测到的 IPF 与流式细胞术观察到的年轻 TO 血小板群体并不完全相同。
本研究旨在评估 HLA I/主要组织相容性 I(MHC I)细胞表面表达作为年轻血小板标志物的可靠性。
通过用 TO 和针对 HLA I/MHC I 分子的抗体对血液进行双重染色,用流式细胞术评估 HLA I/MHC I 表达。
我们发现,具有更高的质膜定位 MHC I 分子表达的血小板显示出增加的 TO 染色和核糖体 P 抗原含量更高。在小鼠的输血实验中,我们表明年轻小鼠血小板表面表达的 MHC I 分子数量在血小板老化过程中减少,在 24 小时内达到基础水平。最后,我们证明对于血小板减少症患者,通过流式细胞术确定 HLA I 表达水平比 TO 染色或使用血液学血液计数器更好地识别年轻血小板。
总的来说,我们的结果强调了 MHC I/HLA I 表达作为识别年轻血小板的有价值参数的相关性。