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Updated: Jul 14, 2025
Infection of Primary Nasal Epithelial Cells Grown at an Air-Liquid Interface to Characterize Human Coronavirus-Host Interactions
Published on: September 22, 2023
Infection of Primary Nasal Epithelial Cells Grown at an Air-Liquid Interface to Characterize Human Coronavirus-Host
Clayton J Otter1, Alejandra Fausto1, Li Hui Tan2
1Department of Microbiology, University of Pennsylvania; Penn Center for Research on Coronaviruses and Other Emerging Pathogens, Perelman School of Medicine, University of Pennsylvania.
Abstract:
Three highly pathogenic human coronaviruses (HCoVs) - SARS-CoV (2002), MERS-CoV (2012), and SARS-CoV-2 (2019) - have emerged and caused significant public health crises in the past 20 years. Four additional HCoVs cause a significant portion of common cold cases each year (HCoV-NL63, -229E, -OC43, and -HKU1), highlighting the importance of studying these viruses in physiologically relevant systems. HCoVs enter the respiratory tract and establish infection in the nasal epithelium, the primary site encountered by all respiratory pathogens. We use a primary nasal epithelial culture system in which patient-derived nasal samples are grown at an air-liquid interface (ALI) to study host-pathogen interactions at this important sentinel site. These cultures recapitulate many features of the in vivo airway, including the cell types present, ciliary function, and mucus production. We describe methods to characterize viral replication, host cell tropism, virus-induced cytotoxicity, and innate immune induction in nasal ALI cultures following HCoV infection, using recent work comparing lethal and seasonal HCoVs as an example1. An increased understanding of host-pathogen interactions in the nose has the potential to provide novel targets for antiviral therapeutics against HCoVs and other respiratory viruses that will likely emerge in the future.
Insights
This study introduces a nasal epithelial air-liquid interface (ALI) culture model to investigate human coronavirus (HCoV) infections in the respiratory tract. This model aids in understanding host-pathogen interactions and developing new antiviral therapies.
Area of Science:
- Virology
- Respiratory Medicine
- Immunology
Background:
- Three highly pathogenic human coronaviruses (HCoVs) — SARS-CoV, MERS-CoV, and SARS-CoV-2 — have caused major public health crises.
- Four common HCoVs (NL63, 229E, OC43, and HKU1) contribute significantly to common cold cases annually.
- HCoVs infect the nasal epithelium, the initial site of respiratory pathogen entry.
Purpose of the Study:
- To establish and utilize a physiologically relevant nasal epithelial air-liquid interface (ALI) culture system.
- To study host-pathogen interactions of HCoVs at the sentinel nasal site.
- To characterize viral replication, tropism, cytotoxicity, and immune responses in nasal ALI cultures.
Main Methods:
- Utilized patient-derived nasal samples cultured at an air-liquid interface (ALI).
- Developed methods to characterize viral replication, host cell tropism, cytotoxicity, and innate immune induction.
- Compared lethal and seasonal HCoVs in the established nasal ALI model.
Main Results:
- Nasal ALI cultures recapitulate key in vivo airway features, including cell types, ciliary function, and mucus production.
- Demonstrated the ability to characterize HCoV infection dynamics, including replication and host responses.
- Provided a platform for comparing different HCoV strains, such as lethal and seasonal types.
Conclusions:
- Nasal ALI cultures serve as a valuable, physiologically relevant model for studying HCoV infections.
- Understanding host-pathogen interactions in the nose can identify novel therapeutic targets.
- This research supports the development of antivirals against current and future emerging respiratory viruses.
