Establishment of Swine Primary Nasal, Tracheal, and Bronchial Epithelial Cell Culture Models for the Study of Influenza Virus Infection

IF 2.2 4区 医学 Q3 BIOCHEMICAL RESEARCH METHODS Journal of virological methods Pub Date : 2024-04-26 DOI:10.1016/j.jviromet.2024.114943
Madelyn Krunkosky , Thomas M. Krunkosky , Victoria Meliopoulos , Constantinos S. Kyriakis , Stacey Schultz-Cherry , S. Mark Tompkins
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Abstract

We established primary porcine nasal, tracheal, and bronchial epithelial cells that recapitulate the physical and functional properties of the respiratory tract and have the ability to fully differentiate. Trans-well cultures demonstrated increased transepithelial electrical resistance over time the presence of tight junctions as demonstrated by immunohistochemistry. The nasal, tracheal, and bronchial epithelial cells developed cilia, secreted mucus, and expressed sialic acids on surface glycoproteins, the latter which are required for influenza A virus infection. Swine influenza viruses were shown to replicate efficiently in the primary epithelial cell cultures, supporting the use of these culture models to assess swine influenza and other virus infection. Primary porcine nasal, tracheal, and bronchial epithelial cell culture models enable assessment of emerging and novel influenza viruses for pandemic potential as well as mechanistic studies to understand mechanisms of infection, reassortment, and generation of novel virus. As swine are susceptible to infection with multiple viral and bacterial respiratory pathogens, these primary airway cell models may enable study of the cellular response to infection by pathogens associated with Porcine Respiratory Disease Complex.

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为研究流感病毒感染建立猪鼻腔、气管和支气管上皮细胞原代培养模型。
我们建立了原代猪鼻、气管和支气管上皮细胞,这些细胞再现了呼吸道的物理和功能特性,并具有完全分化的能力。经孔培养显示,随着时间的推移,经上皮细胞的电阻增加,免疫组化显示存在紧密连接。鼻腔、气管和支气管上皮细胞出现纤毛,分泌粘液,并在表面糖蛋白上表达硅酸,后者是甲型流感病毒感染所必需的。猪流感病毒在原代上皮细胞培养物中能有效复制,这支持使用这些培养模型来评估猪流感和其他病毒感染。原代猪鼻、气管和支气管上皮细胞培养模型可用于评估新出现的新型流感病毒的大流行潜力,也可用于机理研究,以了解感染、重组和产生新型病毒的机制。由于猪易感染多种病毒和细菌性呼吸道病原体,这些原发性气道细胞模型可用于研究猪呼吸道疾病综合征相关病原体感染的细胞反应。
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来源期刊
CiteScore
5.80
自引率
0.00%
发文量
209
审稿时长
41 days
期刊介绍: The Journal of Virological Methods focuses on original, high quality research papers that describe novel and comprehensively tested methods which enhance human, animal, plant, bacterial or environmental virology and prions research and discovery. The methods may include, but not limited to, the study of: Viral components and morphology- Virus isolation, propagation and development of viral vectors- Viral pathogenesis, oncogenesis, vaccines and antivirals- Virus replication, host-pathogen interactions and responses- Virus transmission, prevention, control and treatment- Viral metagenomics and virome- Virus ecology, adaption and evolution- Applied virology such as nanotechnology- Viral diagnosis with novelty and comprehensive evaluation. We seek articles, systematic reviews, meta-analyses and laboratory protocols that include comprehensive technical details with statistical confirmations that provide validations against current best practice, international standards or quality assurance programs and which advance knowledge in virology leading to improved medical, veterinary or agricultural practices and management.
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