Conjunctival epithelial cells resist productive SARS-CoV-2 infection

  1. Robert M. Jackson
  2. Catherine F. Hatton
  3. Jarmila Stremenova Spegarova
  4. Maria Georgiou
  5. Joseph Collin
  6. Emily Stephenson
  7. Bernard Verdon
  8. Iram J. Haq
  9. Rafiqul Hussain
  10. Jonathan M. Coxhead
  11. Hardeep-Singh Mudhar
  12. Bart Wagner
  13. Megan Hasoon
  14. Tracey Davey
  15. Paul Rooney
  16. C.M. Anjam Khan
  17. Chris Ward
  18. Malcolm Brodlie
  19. Muzlifah Haniffa
  20. Sophie Hambleton
  21. Lyle Armstrong
  22. Francisco Figueiredo
  23. Rachel Queen
  24. Christopher J.A. Duncan
  25. Majlinda Lako

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Abstract

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  1. Version published to 10.1016/j.stemcr.2022.05.017
  2. ScreenIT

    SciScore for 10.1101/2021.12.20.473523: (What is this?)

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

    Table 1: Rigor

    EthicsConsent: Human tissue donation: Adult human eyes from three female donors of 52, 78, and 80 years old were donated for research following informed consent.
    Sex as a biological variableHuman tissue donation: Adult human eyes from three female donors of 52, 78, and 80 years old were donated for research following informed consent.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Experimental Models: Cell Lines
    SentencesResources
    Infection of conjunctival ALI cultures with SARS-CoV-2: A clinical isolate from Public Health England of SARS-CoV-2 (BetaCoV/England/2/2020) virus was propagated once in Vero E6 cells.
    Vero E6
    suggested: RRID:CVCL_XD71)
    Software and Algorithms
    SentencesResources
    Statistical Analysis: Statistical analysis was done using GraphPad Prism (version 9.0.0 121).
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)
    Image acquisition was captured through Zen software.
    Zen
    suggested: None
    The DoubletFinder package (McGinnis et al., 2019) was used to find and remove doublets.
    DoubletFinder
    suggested: (DoubletFinder, RRID:SCR_018771)

    Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).

    Results from LimitationRecognizer: We detected the following sentences addressing limitations in the study:
    The ALI organotypic conjunctival model reported in this study overcomes these limitations, as it can be generated in large numbers and comprises all the key conjunctival cell types, including mucin secreting cells, which play an important role in the ocular surface defence against viruses (Mantelli and Argüeso, 2008). Importantly, glycocalyx, a layer of glycolipids and glycoproteins, forming a barrier between the apical surface of the ALI model and the surrounding was observed, mimicking the native barrier of conjunctival epithelium. Consistent with our previous single-cell studies showing expression of relevant entry receptors ACE2 and TMPRSS2, in approximately 6.6% of conjunctival epithelial cells ex vivo (Collin et al., 2021b), SARS-CoV-2 infection in this ALI conjunctival epithelium model indicated broad but relatively low tropism of SARS-COV-2 for the various cell types (epithelial progenitors, basal and superficial conjunctival epithelium, fibroblasts). Importantly, our data showed no evidence of productive replication in the conjunctival epithelium, consistent with recent findings in the corneal epithelium (Miner et al., 2020). Reasons for this post-entry restriction in different ocular surface cell types remain to be defined, but contrast with permissiveness of the conjunctival epithelium to other respiratory viruses (Belser et al., 2012). These data are consistent with the apparently paradoxical detection of SARS-CoV-2 nucleic acid or protein in post-mortem tissue sa...

    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: Please consider improving the rainbow (“jet”) colormap(s) used on pages 32, 33, 34 and 39. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.

    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.

    Results from scite Reference Check: We found no unreliable references.

    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.

  3. Version published to 10.1101/2021.12.20.473523 on bioRxiv