Effects of respiratory syncytial virus infection and major basic protein derived from eosinophils in pulmonary alveolar epithelial cells (A549)
Effects of respiratory syncytial virus infection and major basic protein derived from eosinophils in pulmonary alveolar epithelial cells (A549)
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DOI：
10.1042/cbi20100255
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发表时间：
2011-05-01
影响因子：
3.9
通讯作者：
Kato, Masahiko
中科院分区：
文献类型：
--
作者：
Ishioka, Taisei;Kimura, Hirokazu;Kato, Masahiko
呼吸道合胞病毒(RSV)引起的肺炎和毛细支气管炎可能与包括哮喘在内的高反应性条件有关。嗜酸性蛋白如MBP(主要碱性蛋白)也可能与哮喘的病理生理有关。为探讨呼吸道合胞病毒(RSV)感染和MBP在肺炎高反应性发病机制中的作用，我们观察了RSV感染和MBP对A549(肺泡上皮)细胞的影响。显微镜下观察A549细胞的细胞病变效应。用流式细胞仪和改良的四甲基偶氮唑蓝([3-(4，5-dimethylthiazol-2-yl)-2，5-diphenyltetrazolium)比色法检测细胞的凋亡和死亡。我们还检测了A549细胞上清液中的15种细胞因子和趋化因子。虽然RSV单独作用不影响A549细胞的CPE，但高浓度的MBP可在24 h内引起细胞死亡，RSV和MBP联合作用可协同诱导细胞死亡。在单纯感染RSV的A549细胞中，GM-CSF(粒细胞-巨噬细胞集落刺激因子)的释放明显高于未感染的对照细胞。MBP单独作用后，IL-2、4、5、7、10、12、13、17、干扰素-γ、GM-CSF、粒细胞集落刺激因子和巨噬细胞炎性蛋白-1β的产生均显著高于对照组。值得注意的是，RSV/MBP处理的细胞中GM-CSF和IL-17的水平显著高于MBP单独处理的细胞。提示MBP可协同促进呼吸道合胞病毒感染的A549细胞释放多种细胞因子和趋化因子，促进细胞死亡，提示MBP可能与呼吸道合胞病毒所致毛细支气管炎/肺炎过敏反应的病理生理机制密切相关。
RSV (respiratory syncytial virus)-induced pneumonia and bronchiolitis may be associated with hyperresponsive conditions, including asthma. Eosinophilic proteins such as MBP (major basic protein) may also be associated with the pathophysiology of asthma. To elucidate the roles of RSV infection and MBP in the pathogenesis of pneumonia with hyperresponsiveness, we investigated the effects of RSV infection and MBP on A549 (alveolar epithelial) cells. CPE (cytopathic effects) in A549 cells were observed by microscopy. Apoptosis and cell death was evaluated by flow cytometric analysis and modified MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. We also measured 15 types of cytokines and chemokines in A549 cell supernatants. Although RSV alone did not affect the CPE of A549, high concentrations of MBP resulted in cell death within 24 h. Combinations of RSV and MBP synergistically induced cell death. In A549 cells infected with RSV alone, the release of GM-CSF (granulocyte-macrophage colony-stimulating factor) was significantly enhanced compared with control cells (no infection). In the cells treated with MBP alone, the production of IL (interleukin)-2, 4, 5, 7, 10, 12, 13, 17, IFN (interferon)-gamma, GM-CSF, G-CSF (granulocyte colony-stimulating factor) and MIP (macrophage inflammatory protein)-1 beta was significantly increased compared with control cells. Notably, the levels of GM-CSF and IL-17 in RSV/MBP-treated cells were significantly higher than those treated with MBP alone. These results suggest that MBP synergistically enhanced the release of various cytokines/chemokines and the cell death of RSV-infected A549 cells, indicating that MBP may be closely associated with the pathophysiology of allergic reactions in bronchiolitis/pneumonia due to RSV.