Tags

Type your tag names separated by a space and hit enter

Infection of human Nasal Epithelial Cells with SARS-CoV-2 and a 382-nt deletion isolate lacking ORF8 reveals similar viral kinetics and host transcriptional profiles.
PLoS Pathog. 2020 12; 16(12):e1009130.PP

Abstract

The novel coronavirus SARS-CoV-2 is the causative agent of Coronavirus Disease 2019 (COVID-19), a global healthcare and economic catastrophe. Understanding of the host immune response to SARS-CoV-2 is still in its infancy. A 382-nt deletion strain lacking ORF8 (Δ382 herein) was isolated in Singapore in March 2020. Infection with Δ382 was associated with less severe disease in patients, compared to infection with wild-type SARS-CoV-2. Here, we established Nasal Epithelial cells (NECs) differentiated from healthy nasal-tissue derived stem cells as a suitable model for the ex-vivo study of SARS-CoV-2 mediated pathogenesis. Infection of NECs with either SARS-CoV-2 or Δ382 resulted in virus particles released exclusively from the apical side, with similar replication kinetics. Screening of a panel of 49 cytokines for basolateral secretion from infected NECs identified CXCL10 as the only cytokine significantly induced upon infection, at comparable levels in both wild-type and Δ382 infected cells. Transcriptome analysis revealed the temporal up-regulation of distinct gene subsets during infection, with anti-viral signaling pathways only detected at late time-points (72 hours post-infection, hpi). This immune response to SARS-CoV-2 was significantly attenuated when compared to infection with an influenza strain, H3N2, which elicited an inflammatory response within 8 hpi, and a greater magnitude of anti-viral gene up-regulation at late time-points. Remarkably, Δ382 induced a host transcriptional response nearly identical to that of wild-type SARS-CoV-2 at every post-infection time-point examined. In accordance with previous results, Δ382 infected cells showed an absence of transcripts mapping to ORF8, and conserved expression of other SARS-CoV-2 genes. Our findings shed light on the airway epithelial response to SARS-CoV-2 infection, and demonstrate a non-essential role for ORF8 in modulating host gene expression and cytokine production from infected cells.

Links

PMC Free PDF

Authors+Show Affiliations

Gamage AM
Programme in Emerging Infectious Diseases, Duke-NUS Medical School, Singapore.
Tan KS
Department of Otolaryngology, Infectious Diseases Translational Research Programme, Yong Loo Lin School of Medicine, National University Health System, National University of Singapore, Singapore.
Chan WOY
Programme in Emerging Infectious Diseases, Duke-NUS Medical School, Singapore.
Liu J
Department of Otolaryngology, Infectious Diseases Translational Research Programme, Yong Loo Lin School of Medicine, National University Health System, National University of Singapore, Singapore.
Tan CW
Programme in Emerging Infectious Diseases, Duke-NUS Medical School, Singapore.
Ong YK
Department of Otolaryngology, Head & Neck Surgery, National University Health System, National University Hospital, Singapore.
Thong M
Department of Otolaryngology, Head & Neck Surgery, National University Health System, National University Hospital, Singapore.
Andiappan AK
Singapore Immunology Network (SIgN), A*STAR, Singapore, Singapore.
Anderson DE
Programme in Emerging Infectious Diseases, Duke-NUS Medical School, Singapore.
Wang Y
Department of Otolaryngology, Infectious Diseases Translational Research Programme, Yong Loo Lin School of Medicine, National University Health System, National University of Singapore, Singapore.
Wang LF
Programme in Emerging Infectious Diseases, Duke-NUS Medical School, Singapore. Singhealth Duke-NUS Global Health Institute, Singapore.

MeSH

COVID-19Chemokine CXCL10Epithelial CellsHost-Pathogen InteractionsHumansKineticsNasal MucosaSARS-CoV-2TranscriptomeViral ProteinsVirus Replication

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

33284849

Citation

Gamage, Akshamal M., et al. "Infection of Human Nasal Epithelial Cells With SARS-CoV-2 and a 382-nt Deletion Isolate Lacking ORF8 Reveals Similar Viral Kinetics and Host Transcriptional Profiles." PLoS Pathogens, vol. 16, no. 12, 2020, pp. e1009130.
Gamage AM, Tan KS, Chan WOY, et al. Infection of human Nasal Epithelial Cells with SARS-CoV-2 and a 382-nt deletion isolate lacking ORF8 reveals similar viral kinetics and host transcriptional profiles. PLoS Pathog. 2020;16(12):e1009130.
Gamage, A. M., Tan, K. S., Chan, W. O. Y., Liu, J., Tan, C. W., Ong, Y. K., Thong, M., Andiappan, A. K., Anderson, D. E., Wang, Y., & Wang, L. F. (2020). Infection of human Nasal Epithelial Cells with SARS-CoV-2 and a 382-nt deletion isolate lacking ORF8 reveals similar viral kinetics and host transcriptional profiles. PLoS Pathogens, 16(12), e1009130. https://doi.org/10.1371/journal.ppat.1009130
Gamage AM, et al. Infection of Human Nasal Epithelial Cells With SARS-CoV-2 and a 382-nt Deletion Isolate Lacking ORF8 Reveals Similar Viral Kinetics and Host Transcriptional Profiles. PLoS Pathog. 2020;16(12):e1009130. PubMed PMID: 33284849.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Infection of human Nasal Epithelial Cells with SARS-CoV-2 and a 382-nt deletion isolate lacking ORF8 reveals similar viral kinetics and host transcriptional profiles. AU - Gamage,Akshamal M, AU - Tan,Kai Sen, AU - Chan,Wharton O Y, AU - Liu,Jing, AU - Tan,Chee Wah, AU - Ong,Yew Kwang, AU - Thong,Mark, AU - Andiappan,Anand K, AU - Anderson,Danielle E, AU - Wang,De Yun, AU - Wang,Lin-Fa, Y1 - 2020/12/07/ PY - 2020/08/31/received PY - 2020/11/09/accepted PY - 2020/12/17/revised PY - 2020/12/8/pubmed PY - 2020/12/29/medline PY - 2020/12/7/entrez SP - e1009130 EP - e1009130 JF - PLoS pathogens JO - PLoS Pathog VL - 16 IS - 12 N2 - The novel coronavirus SARS-CoV-2 is the causative agent of Coronavirus Disease 2019 (COVID-19), a global healthcare and economic catastrophe. Understanding of the host immune response to SARS-CoV-2 is still in its infancy. A 382-nt deletion strain lacking ORF8 (Δ382 herein) was isolated in Singapore in March 2020. Infection with Δ382 was associated with less severe disease in patients, compared to infection with wild-type SARS-CoV-2. Here, we established Nasal Epithelial cells (NECs) differentiated from healthy nasal-tissue derived stem cells as a suitable model for the ex-vivo study of SARS-CoV-2 mediated pathogenesis. Infection of NECs with either SARS-CoV-2 or Δ382 resulted in virus particles released exclusively from the apical side, with similar replication kinetics. Screening of a panel of 49 cytokines for basolateral secretion from infected NECs identified CXCL10 as the only cytokine significantly induced upon infection, at comparable levels in both wild-type and Δ382 infected cells. Transcriptome analysis revealed the temporal up-regulation of distinct gene subsets during infection, with anti-viral signaling pathways only detected at late time-points (72 hours post-infection, hpi). This immune response to SARS-CoV-2 was significantly attenuated when compared to infection with an influenza strain, H3N2, which elicited an inflammatory response within 8 hpi, and a greater magnitude of anti-viral gene up-regulation at late time-points. Remarkably, Δ382 induced a host transcriptional response nearly identical to that of wild-type SARS-CoV-2 at every post-infection time-point examined. In accordance with previous results, Δ382 infected cells showed an absence of transcripts mapping to ORF8, and conserved expression of other SARS-CoV-2 genes. Our findings shed light on the airway epithelial response to SARS-CoV-2 infection, and demonstrate a non-essential role for ORF8 in modulating host gene expression and cytokine production from infected cells. SN - 1553-7374 UR - https://www.unboundmedicine.com/medline/citation/33284849/Infection_of_human_Nasal_Epithelial_Cells_with_SARS_CoV_2_and_a_382_nt_deletion_isolate_lacking_ORF8_reveals_similar_viral_kinetics_and_host_transcriptional_profiles_ DB - PRIME DP - Unbound Medicine ER -