Infections of emerging and reemerging viruses (SARS-CoVs, influenza H1N1, etc.) largely and globally affect human health. Animal models often fail to reflect a physiological status because of species tropism of virus infection. Conventional cell lines are usually genetically and phenotypically different from primary cells. Developing an in vitro physiological model to study the infection of emerging viruses will facilitate our understanding of virus-host cell interactions, thereby benefiting antiviral drug discovery. In the current work, we first established normal airway epithelial cells (upper and lower airway track) in 2D and 3D culture systems using conditional reprogramming (CR) and air-liquid interface (ALI) techniques. These long-term cultures maintained differentiation potential. More importantly, these cells express two types of influenza virus receptors, α2-6-Gal- and α2-3-Gal-linked sialic acids, and angiotensin-converting enzyme 2 (ACE2), a receptor for SARS-CoVs as well. These cells were permissive to the infection of pandemic influenza H1N1 (H1N1pdm). In contrast, the lung cancer cell line A549 and immortalized airway epithelial cells (16HBE) were not susceptible to H1N1 infection. A virus-induced cytopathic effect (CPE) on 2D CRC cultures developed in a time-dependent manner. The pathological effects were also readily observed spreading from the apical layer to the basal layer of the 3D ALI culture. This integrated 2D CRC and 3D ALI cultures provide a physiological and personalized in vitro model to study the infection of emerging viruses. This novel model can be used for studying virus biology and host response to viral infection and for antiviral drug discovery.

中文翻译：

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耦合的CRC 2D和ALI 3D培养物可表达新兴病毒的受体，与常规细胞系相比，更适合研究病毒感染。

新兴和新兴病毒（SARS冠状病毒，H1N1流感等）的感染在很大程度上影响全球健康。由于病毒感染的物种趋向性，动物模型通常无法反映生理状态。常规细胞系通常在遗传和表型上与原代细胞不同。体外开发研究新兴病毒感染的生理模型将有助于我们了解病毒与宿主细胞的相互作用，从而有利于抗病毒药物的发现。在目前的工作中，我们首先使用条件重编程（CR）和气液界面（ALI）技术在2D和3D培养系统中建立了正常的气道上皮细胞（上，下气道轨道）。这些长期的文化保持了分化的潜力。更重要的是，这些细胞表达两种类型的流感病毒受体，α 2-6-GAL-和α2-3-Gal连接的唾液酸，以及血管紧张素转化酶2（ACE2），也是SARS-CoV的受体。这些细胞被允许感染大流行性流感H1N1（H1N1pdm）。相反，肺癌细胞系A549和永生化气道上皮细胞（16HBE）对H1N1感染不敏感。对二维CRC培养物的病毒诱导的细胞病变效应（CPE）以时间依赖性方式发展。还很容易观察到病理效应从3D ALI培养的根尖层扩散到基底层。这种集成的2D CRC和3D ALI文化提供了生理和个性化的体外模型，用于研究新兴病毒的感染。这种新颖的模型可用于研究病毒生物学和宿主对病毒感染的反应以及抗病毒药物的发现。