Department of Biological Sciences, University of Idaho, Moscow, ID 83844, USA.
Biological Sciences Division, Pacific Northwest National Laboratory, Richland, WA 99352, USA.
Virology. 2015 Sep;483:96-107. doi: 10.1016/j.virol.2015.03.045. Epub 2015 May 15.
Infection of type II alveolar epithelial (ATII) cells by influenza A viruses (IAV) correlates with severe respiratory disease in humans and mice. To understand pathogenic mechanisms during IAV infection of ATII cells, murine ATII cells were cultured to maintain a differentiated phenotype, infected with IAV-PR8, which causes severe lung pathology in mice, and proteomics analyses were performed using liquid chromatography-mass spectrometry. PR8 infection increased levels of proteins involved in interferon signaling, antigen presentation, and cytoskeleton regulation. Proteins involved in mitochondrial membrane permeability, energy metabolism, and chromatin formation had reduced levels in PR8-infected cells. Phenotypic markers of ATII cells in vivo were identified, confirming the differentiation status of the cultures. Surfactant protein B had decreased levels in PR8-infected cells, which was confirmed by immunoblotting and immunofluorescence assays. Analysis of ATII cell protein profiles will elucidate cellular processes in IAV pathogenesis, which may provide insight into potential therapies to modulate disease severity.
流感 A 病毒 (IAV) 感染 II 型肺泡上皮 (ATII) 细胞与人类和小鼠的严重呼吸道疾病相关。为了了解 IAV 感染 ATII 细胞过程中的发病机制，用液体色谱-质谱联用技术对维持分化表型的原代培养的鼠 ATII 细胞进行了 IAV-PR8 感染的蛋白质组学分析，IAV-PR8 可导致小鼠肺部严重病理改变。PR8 感染增加了干扰素信号、抗原呈递和细胞骨架调节相关蛋白的水平。PR8 感染细胞中线粒体膜通透性、能量代谢和染色质形成相关蛋白的水平降低。鉴定了体内 ATII 细胞的表型标志物，证实了培养物的分化状态。免疫印迹和免疫荧光分析证实，PR8 感染细胞中表面活性剂蛋白 B 的水平降低。对 ATII 细胞蛋白质谱的分析将阐明 IAV 发病机制中的细胞过程，这可能为调节疾病严重程度的潜在治疗方法提供思路。
