Elevated temperature inhibits SARS-CoV-2 replication in respiratory epithelium independently of the induction of IFN-mediated innate immune defences

  1. Vanessa Herder
  2. Kieran Dee
  3. Joanna K. Wojtus
  4. Daniel Goldfarb
  5. Christoforos Rozario
  6. Quan Gu
  7. Ruth F. Jarrett
  8. Ilaria Epifano
  9. Andrew Stevenson
  10. Steven McFarlane
  11. Meredith E. Stewart
  12. Agnieszka M. Szemiel
  13. Rute M. Pinto
  14. Andreu Masdefiol Garriga
  15. Sheila V. Graham
  16. Pablo R. Murcia
  17. Chris Boutell

Reviewed by

Read the full article See related articles

Listed in

Log in to save this article

Abstract

The pandemic spread of SARS-CoV-2, the etiological agent of COVID-19, represents a significant and ongoing international health crisis. A key symptom of SARS-CoV-2 infection is the onset of fever, with a hyperthermic temperature range of 38 to 41°C. Fever is an evolutionarily conserved host response to microbial infection and inflammation that can influence the outcome of viral pathogenicity and regulation of host innate and adaptive immune responses. However, it remains to be determined what effect elevated temperature has on SARS-CoV-2 tropism and replication. Utilizing a 3D air-liquid interface (ALI) model that closely mimics the natural tissue physiology and cellular tropism of SARS-CoV-2 infection in the respiratory airway, we identify tissue temperature to play an important role in the regulation of SARS-CoV-2 infection. We show that temperature elevation induces wide-spread transcriptome changes that impact upon the regulation of multiple pathways, including epigenetic regulation and lncRNA expression, without disruption of general cellular transcription or the induction of interferon (IFN)-mediated antiviral immune defences. Respiratory tissue incubated at temperatures >37°C remained permissive to SARS-CoV-2 infection but severely restricted the initiation of viral transcription, leading to significantly reduced levels of intraepithelial viral RNA accumulation and apical shedding of infectious virus. To our knowledge, we present the first evidence that febrile temperatures associated with COVID-19 inhibit SARS-CoV-2 replication. Our data identify an important role for temperature elevation in the epithelial restriction of SARS-CoV-2 that occurs independently of the induction of canonical IFN-mediated antiviral immune defences and interferon-stimulated gene (ISG) expression.

Article activity feed

  1. ScreenIT

    SciScore for 10.1101/2020.12.04.411389: (What is this?)

    Please note, not all rigor criteria are appropriate for all manuscripts.

    Table 1: Rigor

    Institutional Review Board Statementnot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variableDifferentiation of respiratory epithelium: Primary human bronchiolar epithelial (HBE) cells from a healthy 63-year-old white Caucasian male (non-smoker) were sourced from Epithelix Sarl (Geneva, Switzerland).
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    ACE2 was detected using a rabbit polyclonal anti-hACE2 antibody (Cell Signalling, 4355S; citrate antigen-retrieval) and EnVision+ anti-rabbit HRP (Agilent, K4003).
    a rabbit polyclonal anti-hACE2 antibody ( Cell Signalling , 4355S; citrate antigen-retrieval )
    suggested: None
    EnVision+ anti-rabbit HRP ( Agilent , K4003)
    suggested: None
    anti-rabbit
    suggested: (Agilent Cat# K4003, RRID:AB_2630375)
    K4003
    suggested: (Agilent Cat# K4003, RRID:AB_2630375)
    Mx1 was detected using a mouse monoclonal anti-Mx1 antibody ((63); EDTA antigen-retrieval).
    anti-Mx1
    suggested: None
    SCV2 nucleocapsid protein was detected using a sheep polyclonal anti-SCV2 N protein antibody (University of Dundee, DA114; EDTA antigen-retrieval).
    anti-SCV2 N protein
    suggested: None
    Secondary antibodies for detection were rabbit anti-sheep AlexaFluor 555 (abcam, ab150182) and rabbit anti-mouse AlexaFluor 488 (Sigma-Aldrich, SAB4600056).
    anti-sheep
    suggested: None
    anti-mouse
    suggested: None
    SAB4600056
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    The virus was passaged three times in VeroE6 cells and genotype sequence confirmed by Illumina sequencing.
    VeroE6
    suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)
    Software and Algorithms
    SentencesResources
    Zen black software (Zeiss) was used for image capture and exporting images, with minimal adjustment (image rotation) in Adobe Photoshop for presentation.
    Adobe Photoshop
    suggested: (Adobe Photoshop, RRID:SCR_014199)
    Tissue section image analysis: Scanned Haematoxylin and Eosin (H&E) stained sections were analyzed using Aperio ImageScope analysis software (Leica).
    ImageScope
    suggested: (ImageScope, RRID:SCR_014311)
    RNA-Seq reads were quality assessed (FastQC; http://www.bioinformatics.babraham.ac.uk/projects/fastqc) and sequence adaptors removed (TrimGalore; https://www.bioinformatics.babraham.ac.uk/projects/trim_galore/).
    https://www.bioinformatics.babraham.ac.uk/projects/trim_galore/
    suggested: (Trim Galore, RRID:SCR_011847)
    HISAT2 is a fast and sensitive splice aware mapper, which aligns RNA sequencing reads to mammalian-sized genomes using FM index strategy (64).
    HISAT2
    suggested: (HISAT2, RRID:SCR_015530)
    RNA-Seq reads were also mapped to SARS-CoV-2 (GISAID accession ID:EPI_ISL_407073) using Bowtie2 (65).
    Bowtie2
    suggested: (Bowtie 2, RRID:SCR_016368)
    The edgeR package was used to calculate the gene expression level and to analyze differentially expressed genes between sample groups (67).
    edgeR
    suggested: (edgeR, RRID:SCR_012802)
    In Metascape, all DEGs were used for differential pathway analysis.
    Metascape
    suggested: (Metascape, RRID:SCR_016620)
    Differential expressed (p<0.05, ≥ 1.5 or ≤ -1.5 log2 FC) lncRNAs and miRNAs were identified using the Ensembl BioMart tool (http://www.ensembl.org/biomart/martview/05285d5f063a05a82b8ba71fe18a0f18).
    Ensembl
    suggested: (Ensembl, RRID:SCR_002344)
    Statistical analysis: GraphPad Prism (version 9) was used for statistical analysis.
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)

    Results from OddPub: Thank you for sharing your data.

    Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.

    Results from TrialIdentifier: No clinical trial numbers were referenced.

    Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

    Results from JetFighter: We did not find any issues relating to colormaps.

    Results from rtransparent:
    • Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
    • Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
    • No protocol registration statement was detected.

    About SciScore

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.

  2. Version published to 10.1101/2020.12.04.411389v1 on bioRxiv