Long-Term Modeling of SARS-CoV-2 Infection of <italic toggle="yes">In Vitro</italic> Cultured Polarized Human Airway Epithelium

ABSTRACT Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) replicates throughout human airways. The polarized human airway epithelium (HAE) cultured at an airway-liquid interface (HAE-ALI) is an in vitro model mimicking the in vivo human mucociliary airway epithelium and supports the repl...

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Autores principales:	Siyuan Hao, Kang Ning, Cagla Aksu Kuz, Kai Vorhies, Ziying Yan, Jianming Qiu
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	SARS-CoV-2 human airway epithelium epithelial damage recurrent infection airway epithelial damage long-term infection Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/9fa28d51b4454c60a53895cd42672c9d
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spelling	oai:doaj.org-article:9fa28d51b4454c60a53895cd42672c9d2021-11-15T15:55:43ZLong-Term Modeling of SARS-CoV-2 Infection of <italic toggle="yes">In Vitro</italic> Cultured Polarized Human Airway Epithelium10.1128/mBio.02852-202150-7511https://doaj.org/article/9fa28d51b4454c60a53895cd42672c9d2020-12-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02852-20https://doaj.org/toc/2150-7511ABSTRACT Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) replicates throughout human airways. The polarized human airway epithelium (HAE) cultured at an airway-liquid interface (HAE-ALI) is an in vitro model mimicking the in vivo human mucociliary airway epithelium and supports the replication of SARS-CoV-2. Prior studies characterized only short-period SARS-CoV-2 infection in HAE. In this study, continuously monitoring the SARS-CoV-2 infection in HAE-ALI cultures for a long period of up to 51 days revealed that SARS-CoV-2 infection was long lasting with recurrent replication peaks appearing between an interval of approximately 7 to 10 days, which was consistent in all the tested HAE-ALI cultures derived from 4 lung bronchi of independent donors. We also identified that SARS-CoV-2 does not infect HAE from the basolateral side, and the dominant SARS-CoV-2 permissive epithelial cells are ciliated cells and goblet cells, whereas virus replication in basal cells and club cells was not detected. Notably, virus infection immediately damaged the HAE, which is demonstrated by dispersed zonula occludens-1 (ZO-1) expression without clear tight junctions and partial loss of cilia. Importantly, we identified that SARS-CoV-2 productive infection of HAE requires a high viral load of >2.5 × 105 virions per cm2 of epithelium. Thus, our studies highlight the importance of a high viral load and that epithelial renewal initiates and maintains a recurrent infection of HAE with SARS-CoV-2. IMPORTANCE The pandemic of coronavirus disease 2019 (COVID-19), which is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has led to >35 million confirmed cases and >1 million fatalities worldwide. SARS-CoV-2 mainly replicates in human airway epithelia in COVID-19 patients. In this study, we used in vitro cultures of polarized human bronchial airway epithelium to model SARS-CoV-2 replication for a period of 21 to 51 days. We discovered that in vitro airway epithelial cultures endure a long-lasting SARS-CoV-2 propagation with recurrent peaks of progeny virus release at an interval of approximately 7 to 10 days. Our study also revealed that SARS-CoV-2 infection causes airway epithelia damage with disruption of tight junction function and loss of cilia. Importantly, SARS-CoV-2 exhibits a polarity of infection in airway epithelium only from the apical membrane; it infects ciliated and goblet cells but not basal and club cells. Furthermore, the productive infection of SARS-CoV-2 requires a high viral load of over 2.5 × 105 virions per cm2 of epithelium. Our study highlights that the proliferation of airway basal cells and regeneration of airway epithelium may contribute to the recurrent infections.Siyuan HaoKang NingCagla Aksu KuzKai VorhiesZiying YanJianming QiuAmerican Society for MicrobiologyarticleSARS-CoV-2human airway epitheliumepithelial damagerecurrent infectionairway epithelial damagelong-term infectionMicrobiologyQR1-502ENmBio, Vol 11, Iss 6 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	SARS-CoV-2 human airway epithelium epithelial damage recurrent infection airway epithelial damage long-term infection Microbiology QR1-502
spellingShingle	SARS-CoV-2 human airway epithelium epithelial damage recurrent infection airway epithelial damage long-term infection Microbiology QR1-502 Siyuan Hao Kang Ning Cagla Aksu Kuz Kai Vorhies Ziying Yan Jianming Qiu Long-Term Modeling of SARS-CoV-2 Infection of <italic toggle="yes">In Vitro</italic> Cultured Polarized Human Airway Epithelium
description	ABSTRACT Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) replicates throughout human airways. The polarized human airway epithelium (HAE) cultured at an airway-liquid interface (HAE-ALI) is an in vitro model mimicking the in vivo human mucociliary airway epithelium and supports the replication of SARS-CoV-2. Prior studies characterized only short-period SARS-CoV-2 infection in HAE. In this study, continuously monitoring the SARS-CoV-2 infection in HAE-ALI cultures for a long period of up to 51 days revealed that SARS-CoV-2 infection was long lasting with recurrent replication peaks appearing between an interval of approximately 7 to 10 days, which was consistent in all the tested HAE-ALI cultures derived from 4 lung bronchi of independent donors. We also identified that SARS-CoV-2 does not infect HAE from the basolateral side, and the dominant SARS-CoV-2 permissive epithelial cells are ciliated cells and goblet cells, whereas virus replication in basal cells and club cells was not detected. Notably, virus infection immediately damaged the HAE, which is demonstrated by dispersed zonula occludens-1 (ZO-1) expression without clear tight junctions and partial loss of cilia. Importantly, we identified that SARS-CoV-2 productive infection of HAE requires a high viral load of >2.5 × 105 virions per cm2 of epithelium. Thus, our studies highlight the importance of a high viral load and that epithelial renewal initiates and maintains a recurrent infection of HAE with SARS-CoV-2. IMPORTANCE The pandemic of coronavirus disease 2019 (COVID-19), which is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has led to >35 million confirmed cases and >1 million fatalities worldwide. SARS-CoV-2 mainly replicates in human airway epithelia in COVID-19 patients. In this study, we used in vitro cultures of polarized human bronchial airway epithelium to model SARS-CoV-2 replication for a period of 21 to 51 days. We discovered that in vitro airway epithelial cultures endure a long-lasting SARS-CoV-2 propagation with recurrent peaks of progeny virus release at an interval of approximately 7 to 10 days. Our study also revealed that SARS-CoV-2 infection causes airway epithelia damage with disruption of tight junction function and loss of cilia. Importantly, SARS-CoV-2 exhibits a polarity of infection in airway epithelium only from the apical membrane; it infects ciliated and goblet cells but not basal and club cells. Furthermore, the productive infection of SARS-CoV-2 requires a high viral load of over 2.5 × 105 virions per cm2 of epithelium. Our study highlights that the proliferation of airway basal cells and regeneration of airway epithelium may contribute to the recurrent infections.
format	article
author	Siyuan Hao Kang Ning Cagla Aksu Kuz Kai Vorhies Ziying Yan Jianming Qiu
author_facet	Siyuan Hao Kang Ning Cagla Aksu Kuz Kai Vorhies Ziying Yan Jianming Qiu
author_sort	Siyuan Hao
title	Long-Term Modeling of SARS-CoV-2 Infection of <italic toggle="yes">In Vitro</italic> Cultured Polarized Human Airway Epithelium
title_short	Long-Term Modeling of SARS-CoV-2 Infection of <italic toggle="yes">In Vitro</italic> Cultured Polarized Human Airway Epithelium
title_full	Long-Term Modeling of SARS-CoV-2 Infection of <italic toggle="yes">In Vitro</italic> Cultured Polarized Human Airway Epithelium
title_fullStr	Long-Term Modeling of SARS-CoV-2 Infection of <italic toggle="yes">In Vitro</italic> Cultured Polarized Human Airway Epithelium
title_full_unstemmed	Long-Term Modeling of SARS-CoV-2 Infection of <italic toggle="yes">In Vitro</italic> Cultured Polarized Human Airway Epithelium
title_sort	long-term modeling of sars-cov-2 infection of <italic toggle="yes">in vitro</italic> cultured polarized human airway epithelium
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/9fa28d51b4454c60a53895cd42672c9d
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Long-Term Modeling of SARS-CoV-2 Infection of <italic toggle="yes">In Vitro</italic> Cultured Polarized Human Airway Epithelium

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