Jurkeviciute Egle, Januskevicius Andrius, Rimkunas Airidas, Palacionyte Jolita, Malakauskas Kestutis
Laboratory of Pulmonology, Department of Pulmonology, Lithuanian University of Health Sciences, LT-44307 Kaunas, Lithuania.
Department of Pulmonology, Lithuanian University of Health Sciences, LT-44307 Kaunas, Lithuania.
J Pers Med. 2021 Aug 24;11(9):829. doi: 10.3390/jpm11090829.
Eosinophilic inflammation is one of the main pathophysiological features in asthma. Two subtypes of eosinophils exist in the lung and systemic circulation: lung-resident eosinophils (rEOS) and inflammatory eosinophils (iEOS). We evaluated the expression of αβ and αβ integrins of eosinophil subtypes and their influence on airway smooth muscle (ASM) cell proliferation and viability in asthma. We included 16 severe non-allergic eosinophilic asthma (SNEA) patients, 13 steroid-free, non-severe allergic asthma (AA) patients, and 12 healthy control subjects (HS). For AA patients, a bronchial allergen challenge with was performed. The eosinophil subtypes were distinguished using magnetic bead-labeled antibodies against surface CD62L, and individual combined cell cultures were prepared with ASM cells. The integrins gene expression was analyzed by a quantitative real-time polymerase chain reaction. Proliferation was assessed by the Alamar blue assay, and viability by annexin V and propidium iodide staining. rEOS-like cells were characterized by the relatively higher gene expression of the β integrin subunit, whereas iEOS-like cells were characterized by the α and β integrin subunits. The inclusion of either eosinophil subtypes in co-culture significantly increased the proliferation of ASM cells, and the effect of rEOS-like cells was stronger than iEOS-like cells ( < 0.05). Furthermore, rEOS-like cells had a more pronounced effect on reducing ASM cell apoptosis compared to that of iEOS-like cells ( < 0.05). Lastly, the bronchial allergen challenge significantly enhanced only the iEOS-like cells' effect on ASM cell proliferation and viability in AA patients ( < 0.05). These findings highlight the different expression of αβ and αβ integrins on distinct eosinophil subtypes in asthma. Therefore, rEOS-like cells have a stronger effect in stimulating ASM cell proliferation and viability; however, contact with specific allergens mainly enhances pro-proliferative iEOS-like cell properties.
嗜酸性粒细胞炎症是哮喘的主要病理生理特征之一。肺和体循环中存在两种嗜酸性粒细胞亚型：肺驻留嗜酸性粒细胞（rEOS）和炎症性嗜酸性粒细胞（iEOS）。我们评估了嗜酸性粒细胞亚型的αβ和αβ整合素表达及其对哮喘气道平滑肌（ASM）细胞增殖和活力的影响。我们纳入了16例重度非过敏性嗜酸性粒细胞性哮喘（SNEA）患者、13例无类固醇的非重度过敏性哮喘（AA）患者和12名健康对照者（HS）。对于AA患者，进行了支气管过敏原激发试验。使用针对表面CD62L的磁珠标记抗体区分嗜酸性粒细胞亚型，并与ASM细胞制备单个联合细胞培养物。通过定量实时聚合酶链反应分析整合素基因表达。通过Alamar蓝测定法评估增殖，通过膜联蛋白V和碘化丙啶染色评估活力。rEOS样细胞的特征是β整合素亚基的基因表达相对较高，而iEOS样细胞的特征是α和β整合素亚基。共培养中加入任何一种嗜酸性粒细胞亚型均显著增加ASM细胞的增殖，且rEOS样细胞的作用强于iEOS样细胞（P<0.05）。此外，与iEOS样细胞相比，rEOS样细胞对减少ASM细胞凋亡的作用更明显（P<0.05）。最后，支气管过敏原激发试验仅显著增强了AA患者中iEOS样细胞对ASM细胞增殖和活力的作用（P<0.05）。这些发现突出了哮喘中不同嗜酸性粒细胞亚型上αβ和αβ整合素的不同表达。因此，rEOS样细胞在刺激ASM细胞增殖和活力方面具有更强的作用；然而，与特定过敏原接触主要增强促增殖的iEOS样细胞特性。
