Vagida M S, Arakelyan A, Lebedeva A M, Grivel J-Ch, Shpektor A V, Vasilieva E Yu, Margolis L B
Moscow State University of Medicine and Dentistry, Laboratory of Atherothrombosis, 109240 Moscow, Russia.
Section on Intercellular Interactions, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, 20892, Bethesda, Maryland, USA.
Biochemistry (Mosc). 2016 Apr;81(4):382-391. doi: 10.1134/S0006297916040088.
Extracellular vesicles (EVs) are released from various cell types and play an important role in intercellular interactions. In our study, we investigated abundance of individual EVs in patients with acute forms of ischemic heart disease. Previously, we developed an approach for individual analysis of EVs conjugated with magnetic nanoparticles (MNPs), which was applied in the current study for analyzing phenotypic composition of EVs (by staining for markers CD31, CD41a, and CD63). EVs were isolated using fluorescently labeled MNPs containing anti-CD31, CD41a, or CD63 antibodies and analyzed by combining fluorescently labeled anti-CD41a and CD63, CD31 and CD63, or CD41a and CD31 antibodies, respectively. EVs were analyzed in 30 individuals: 17 healthy volunteers and 13 patients with acute coronary syndrome (ACS). Six and seven ACS patients were with acute myocardial infarction and unstable angina, respectively. It was found that patients with ACS and healthy volunteers contained a dominant subset of EVs expressing surface CD41a antigen, suggesting that they originated from platelets. In addition, the total number of EVs isolated using either of the surface markers examined in our study was higher in patients with ACS compared to healthy volunteers. The subgroup of patients with acute myocardial infarction was found to contain significantly higher number of blood EVs compared to the control group. Moreover, increased number of EVs in patients with ACS is mainly due to the increased number of EVs in the subset of EVs bearing CD41a. By analyzing individual EVs, we found that plasma of patients with ACS, particularly upon developing of myocardial infarction, contained dominant platelet-derived EVs fraction, which may reflect activation of platelets in such patients.
细胞外囊泡(EVs)由多种细胞类型释放,在细胞间相互作用中发挥重要作用。在我们的研究中,我们调查了急性缺血性心脏病患者个体EVs的丰度。此前,我们开发了一种用于与磁性纳米颗粒(MNPs)结合的EVs个体分析方法,该方法在本研究中用于分析EVs的表型组成(通过对CD31、CD41a和CD63标志物进行染色)。使用含有抗CD31、CD41a或CD63抗体的荧光标记MNPs分离EVs,并分别通过结合荧光标记的抗CD41a和CD63、CD31和CD63或CD41a和CD31抗体进行分析。对30名个体的EVs进行了分析:17名健康志愿者和13名急性冠状动脉综合征(ACS)患者。分别有6名和7名ACS患者患有急性心肌梗死和不稳定型心绞痛。结果发现,ACS患者和健康志愿者中都含有表达表面CD41a抗原的EVs优势亚群,表明它们起源于血小板。此外,与健康志愿者相比,使用我们研究中检测的任何一种表面标志物分离的EVs总数在ACS患者中更高。发现急性心肌梗死患者亚组的血液EVs数量明显高于对照组。此外,ACS患者中EVs数量增加主要是由于携带CD41a的EVs亚群数量增加。通过分析个体EVs,我们发现ACS患者的血浆,尤其是在发生心肌梗死后,含有占主导地位的血小板衍生EVs部分,这可能反映了此类患者血小板的激活。