Cañas José Antonio, Sastre Beatriz, Mazzeo Carla, Fernández-Nieto Mar, Rodrigo-Muñoz José Manuel, González-Guerra Andrés, Izquierdo Manuel, Barranco Pilar, Quirce Santiago, Sastre Joaquín, Del Pozo Victoria
Department of Immunology, IIS-Fundación Jiménez Díaz, Madrid, Spain.
CIBER de Enfermedades Respiratorias (CIBERES), Madrid, Spain.
J Leukoc Biol. 2017 May;101(5):1191-1199. doi: 10.1189/jlb.3AB0516-233RR. Epub 2017 Jan 17.
Eosinophils are able to secrete exosomes that have an undefined role in asthma pathogenesis. We hypothesized that exosomes released by eosinophils autoregulate and promote eosinophil function. Eosinophils of patients with asthma ( = 58) and healthy volunteers ( = 16) were purified from peripheral blood, and exosomes were isolated and quantified from eosinophils of the asthmatic and healthy populations. Apoptosis, adhesion, adhesion molecules expression, and migration assays were performed with eosinophils in the presence or absence of exosomes from healthy and asthmatic individuals. Reactive oxygen species (ROS) were evaluated by flow cytometry with an intracellular fluorescent probe and nitric oxide (NO) and a colorimetric kit. In addition, exosomal proteins were analyzed by mass spectrometry. Eosinophil-derived exosomes induced an increase in NO and ROS production on eosinophils. Moreover, exosomes could act as a chemotactic factor on eosinophils, and they produced an increase in cell adhesion, giving rise to a specific augmentation of adhesion molecules, such as ICAM-1 and integrin α2. Protein content between exosomes from healthy and asthmatic individuals seems to be similar in both groups. In conclusion, we found that exosomes from the eosinophils of patients with asthma could modify several specific eosinophil functions related to asthma pathogenesis and that they could contribute fundamentally to the development and maintenance of asthma.
嗜酸性粒细胞能够分泌在哮喘发病机制中作用尚不明确的外泌体。我们推测嗜酸性粒细胞释放的外泌体可自我调节并促进嗜酸性粒细胞功能。从哮喘患者(n = 58)和健康志愿者(n = 16)的外周血中纯化嗜酸性粒细胞,并从哮喘患者和健康人群的嗜酸性粒细胞中分离并定量外泌体。在有或没有来自健康个体和哮喘个体的外泌体存在的情况下,对嗜酸性粒细胞进行凋亡、黏附、黏附分子表达和迁移分析。使用细胞内荧光探针通过流式细胞术评估活性氧(ROS),并使用比色试剂盒评估一氧化氮(NO)。此外,通过质谱分析外泌体蛋白。嗜酸性粒细胞衍生的外泌体可诱导嗜酸性粒细胞产生的NO和ROS增加。此外,外泌体可作为嗜酸性粒细胞的趋化因子,它们会增加细胞黏附,导致黏附分子如细胞间黏附分子-1(ICAM-1)和整合素α2的特异性增加。健康个体和哮喘个体的外泌体之间的蛋白质含量在两组中似乎相似。总之,我们发现哮喘患者嗜酸性粒细胞来源的外泌体可改变与哮喘发病机制相关的几种特定嗜酸性粒细胞功能,并且它们可能从根本上促进哮喘的发生和维持。
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