College of Pulmonary & Critical Care Medicine, 8th Medical Center, Chinese PLA General Hospital, 100091 Beijing, China.
Department of Critical Care Medicine, Chinese PLA Medical School, 100853 Beijing, China.
Discov Med. 2023 Aug;35(177):539-552. doi: 10.24976/Discov.Med.202335177.55.
Influenza is an important respiratory tract pathogen that causes substantial seasonal and pandemic morbidity and mortality. The aim of this study was to systematically analyze the transcriptome characteristics of peripheral blood mononuclear cells (PBMCs) after influenza A virus infection by constructing a human lung microarray model composed of PBMCs to simulate the influenza A virus infection process.
A human lung microarray model was constructed using alveolar epithelial cells, vascular endothelial cells, alveolar macrophages and PBMCs, for simulation of the process of influenza A virus infection. The transcriptome characteristics of PBMCs after influenza A virus infection were analyzed by a single-cell RNA sequencing system.
The study could realistically mimic the structure and physiological functions of the alveoli using immunofluorescence staining and expression of the specific marker. After the influenza A virus infected the upper lung chip channels, the epithelial cells underwent a high inflammatory response and spread to endothelial cells. Under experimental conditions, the Influenza A virus infection did not compromise the integrity of epithelial cells, but caused damage to endothelial cells and barrier dysfunction. Single-cell RNA sequencing of PBMCs showed that B and cluster of differentiation 4 (CD4) T cells played important immunomodulatory roles in response to influenza A virus infection, including significantly activating type I interferon signaling pathway, regulating cytokine and chemokine signaling pathway. Especially genes involved in cellular communication were significantly highly expressed post-infection.
All these results suggested that the interactions among immune cells played a crucial role in endothelial cell injury and immune cell recruitment after influenza virus infection. This lung-on-chip infection model combined with single-cell RNA sequencing provided a unique platform that can closely investigate the lung immune response to influenza A virus infection and new therapeutic strategies for influenza.
流感是一种重要的呼吸道病原体，可导致大量季节性和大流行性发病和死亡。本研究旨在通过构建由外周血单个核细胞（PBMC）组成的人肺微阵列模型来模拟流感病毒感染过程，系统分析流感病毒感染后 PBMC 的转录组特征。
使用肺泡上皮细胞、血管内皮细胞、肺泡巨噬细胞和 PBMC 构建人肺微阵列模型，模拟流感病毒感染过程。通过单细胞 RNA 测序系统分析流感病毒感染后 PBMC 的转录组特征。
通过免疫荧光染色和特定标志物的表达，本研究能够真实地模拟肺泡的结构和生理功能。流感病毒感染上肺芯片通道后，上皮细胞发生强烈的炎症反应，并扩散到内皮细胞。在实验条件下，流感病毒感染并不损害上皮细胞的完整性，但会导致内皮细胞损伤和屏障功能障碍。PBMC 的单细胞 RNA 测序显示，B 细胞和 CD4+T 细胞在应对流感病毒感染时发挥重要的免疫调节作用，包括显著激活 I 型干扰素信号通路，调节细胞因子和趋化因子信号通路。感染后细胞间通讯相关基因的表达明显上调。
这些结果表明，免疫细胞之间的相互作用在流感病毒感染后内皮细胞损伤和免疫细胞募集中起关键作用。该肺芯片感染模型结合单细胞 RNA 测序提供了一个独特的平台，可以深入研究肺对流感病毒感染的免疫反应和流感的新治疗策略。
