National Institute for Viral Disease Control and Prevention, China CDC, Key Laboratory for Medical Virology National Health Commission, 100 Ying-Xin St., Xi-Cheng District, Beijing, 100052, China.
The First Hospital of Lanzhou University, Lanzhou, 730000, China.
Virol J. 2019 Apr 3;16(1):44. doi: 10.1186/s12985-019-1146-4.
Since H7N9 influenza A virus (H7N9) was first reported in 2013, five waves of outbreaks have occurred, posing a huge threat to human health. In preparation for a potential H7N9 epidemic, it is essential to evaluate the efficacy of anti-H7N9 drugs with an appropriate model.
Well-differentiated pseudostratified human airway epithelium (HAE) cells were grown at the air-liquid interface, and the H7N9 cell tropism and cytopathic effect were detected by immunostaining and hematoxylin-eosin (HE) staining. The H7N9 replication kinetics and anti-H7N9 effect of recombinant human α2b (rhIFN-α2b) and rhIFN-λ1 were compared with different cell lines. The H7N9 viral load and interferon-stimulated gene (ISG) expression were quantified by real-time PCR assays.
H7N9 could infect both ciliated and non-ciliated cells within the three-dimensional (3D) HAE cell culture, which reduced the number of cilia and damaged the airways. The H7N9 replication kinetics differed between traditional cells and 3D HAE cells. Interferon had antiviral activity against H7N9 and alleviated epithelial cell lesions; the antiviral activity of rhIFN-α2b was slightly better than that of rhIFN-λ1. In normal cells, rhIFN-α2b induced a greater amount of ISG expression (MX1, OAS1, IFITM3, and ISG15) compared with rhIFN-λ1, but in 3D HAE cells, this trend was reversed.
Both rhIFN-α2b and rhIFN-λ1 had antiviral activity against H7N9, and this protection was related to the induction of ISGs. The 3D cell culture model is suitable for evaluating interferon antiviral activity because it can demonstrate realistic in vivo-like effects.
自 2013 年首次报告 H7N9 甲型流感病毒（H7N9）以来，已发生了五波疫情爆发，对人类健康构成了巨大威胁。为应对潜在的 H7N9 疫情，必须使用适当的模型来评估抗 H7N9 药物的疗效。
在气液界面培养分化良好的人假复层呼吸道上皮（HAE）细胞，通过免疫染色和苏木精-伊红（HE）染色检测 H7N9 的细胞嗜性和细胞病变效应。比较重组人α2b（rhIFN-α2b）和 rhIFN-λ1 在不同细胞系中的 H7N9 复制动力学和抗 H7N9 作用。通过实时 PCR 检测 H7N9 病毒载量和干扰素刺激基因（ISG）表达。
H7N9 可感染三维（3D）HAE 细胞培养物中的纤毛细胞和非纤毛细胞，从而减少纤毛数量并损害气道。H7N9 的复制动力学在传统细胞和 3D HAE 细胞之间存在差异。干扰素对 H7N9 具有抗病毒活性，并减轻了上皮细胞损伤；rhIFN-α2b 的抗病毒活性略优于 rhIFN-λ1。在正常细胞中，rhIFN-α2b 诱导的 ISG 表达（MX1、OAS1、IFITM3 和 ISG15）量大于 rhIFN-λ1，但在 3D HAE 细胞中，这种趋势则相反。
rhIFN-α2b 和 rhIFN-λ1 对 H7N9 均具有抗病毒活性，这种保护作用与诱导 ISGs 有关。3D 细胞培养模型适合评估干扰素的抗病毒活性，因为它可以模拟真实的体内效果。
