Department of Internal Medicine I, School of Medicine, University hospital rechts der Isar, Technical University of Munich, Munich, Germany.
German Center for Cardiovascular Research (DZHK), Partner Site Munich Heart Alliance, Munich, Germany.
Cell Death Dis. 2021 Jan 5;12(1):50. doi: 10.1038/s41419-020-03333-9.
Novel coronavirus disease 2019 (COVID-19) is associated with a hypercoagulable state, characterized by abnormal coagulation parameters and by increased incidence of cardiovascular complications. With this study, we aimed to investigate the activation state and the expression of transmembrane proteins in platelets of hospitalized COVID-19 patients. We investigated transmembrane proteins expression with a customized mass cytometry panel of 21 antibodies. Platelets of 8 hospitalized COVID-19 patients not requiring intensive care support and without pre-existing conditions were compared to platelets of healthy controls (11 donors) with and without in vitro stimulation with thrombin receptor-activating peptide (TRAP). Mass cytometry of non-stimulated platelets detected an increased surface expression of activation markers P-Selectin (0.67 vs. 1.87 median signal intensity for controls vs. patients, p = 0.0015) and LAMP-3 (CD63, 0.37 vs. 0.81, p = 0.0004), the GPIIb/IIIa complex (4.58 vs. 5.03, p < 0.0001) and other adhesion molecules involved in platelet activation and platelet-leukocyte interactions. Upon TRAP stimulation, mass cytometry detected a higher expression of P-selectin in COVID-19 samples compared to controls (p < 0.0001). However, we observed a significantly reduced capacity of COVID-19 platelets to increase the expression of activation markers LAMP-3 and P-Selectin upon stimulation with TRAP. We detected a hyperactivated phenotype in platelets during SARS-CoV-2 infection, consisting of highly expressed platelet activation markers, which might contribute to the hypercoagulopathy observed in COVID-19. In addition, several transmembrane proteins were more highly expressed compared to healthy controls. These findings support research projects investigating antithrombotic and antiplatelet treatment regimes in COVID-19 patients, and provide new insights on the phenotypical platelet expression during SARS-CoV-2 infection.
新型冠状病毒病 2019(COVID-19)与高凝状态有关,其特征为凝血参数异常和心血管并发症发生率增加。本研究旨在探究住院 COVID-19 患者血小板的激活状态和跨膜蛋白的表达。我们使用定制的 21 抗体质谱流式细胞术面板研究跨膜蛋白的表达。与未接受重症监护支持且无预先存在疾病的 8 名住院 COVID-19 患者的血小板相比,我们比较了健康对照者(11 名供体)和体外用血栓酶受体激活肽(TRAP)刺激后的血小板。未刺激血小板的质谱流式细胞术检测到活化标志物 P-选择素(P-Selectin)(0.67 对 1.87 中位数信号强度,对照者对患者,p=0.0015)和 LAMP-3(CD63,0.37 对 0.81,p=0.0004)、GPIIb/IIIa 复合物(4.58 对 5.03,p<0.0001)和其他参与血小板激活和血小板-白细胞相互作用的黏附分子的表面表达增加。TRAP 刺激后,质谱流式细胞术检测到 COVID-19 样本中的 P-选择素表达高于对照者(p<0.0001)。然而,我们观察到 COVID-19 血小板在 TRAP 刺激下增加活化标志物 LAMP-3 和 P-选择素表达的能力显著降低。我们在 SARS-CoV-2 感染期间检测到血小板的高激活表型,包括高度表达的血小板活化标志物,这可能导致 COVID-19 中观察到的高凝状态。此外,与健康对照者相比,几种跨膜蛋白的表达更高。这些发现支持针对 COVID-19 患者的抗血栓和抗血小板治疗方案的研究项目,并为 SARS-CoV-2 感染期间血小板的表型表达提供新的见解。