Ocular conjunctival inoculation of SARS-CoV-2 can cause mild COVID-19 in rhesus macaques

  1. Wei Deng
  2. Linlin Bao
  3. Hong Gao
  4. Zhiguang Xiang
  5. Yajin Qu
  6. Zhiqi Song
  7. Shuran Gong
  8. Jiayi Liu
  9. Jiangning Liu
  10. Pin Yu
  11. Feifei Qi
  12. Yanfeng Xu
  13. Fengli Li
  14. Chong Xiao
  15. Qi Lv
  16. Jing Xue
  17. Qiang Wei
  18. Mingya Liu
  19. Guanpeng Wang
  20. Shunyi Wang
  21. Haisheng Yu
  22. Ting Chen
  23. Xing Liu
  24. Wenjie Zhao
  25. Yunlin Han
  26. Chuan Qin

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Abstract

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is highly transmitted through the respiratory route, but potential extra-respiratory routes of SARS-CoV-2 transmission remain uncertain. Here we inoculated five rhesus macaques with 1 × 10 6 TCID 50 of SARS-CoV-2 conjunctivally (CJ), intratracheally (IT), and intragastrically (IG). Nasal and throat swabs collected from CJ and IT had detectable viral RNA at 1–7 days post-inoculation (dpi). Viral RNA was detected in anal swabs from only the IT group at 1–7 dpi. Viral RNA was undetectable in tested swabs and tissues after intragastric inoculation. The CJ infected animal had a higher viral load in the nasolacrimal system than the IT infected animal but also showed mild interstitial pneumonia, suggesting distinct virus distributions. This study shows that infection via the conjunctival route is possible in non-human primates; further studies are necessary to compare the relative risk and pathogenesis of infection through these different routes in more detail.

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  1. Version published to 10.1038/s41467-020-18149-6
  2. ScreenIT

    SciScore for 10.1101/2020.03.13.990036: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIACUC: The Institutional Animal Care and Use Committee of the Institute of Laboratory Animal Science, Peking Union Medical College, reviewed and authorized all the programs in this research including animals (BLL20001).
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variableAnimal experiments: Five male rhesus macaques (Macaca Mulatta) between the ages of 3 and 5 years were used in this research.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    All sera were collected on 0, 7, 14 and 21 dpi for serologic detection to exam whether there presence the IgG antibodies reactive with SARS-CoV-2 antigens.
    IgG
    suggested: None
    ELISA antibody assay: Sera of Each animal were collected to detect the SARS-CoV-2 antibody through enzyme-linked immunosorbent assay (ELISA) on 0, 7, 14 and 21 dpi.
    SARS-CoV-2
    suggested: None
    Sera samples were diluted at 1:100, and then were added to different wells and maintained at 37 °C for 30 minutes, followed by the goat anti-monkey antibody labeled with horseradish peroxidase (Abcam, ab112767) incubated at room temperature for 30 minutes.
    anti-monkey
    suggested: (Abcam Cat# ab112767, RRID:AB_10866625)
    HRP-labeled goat anti-mouse IgG secondary antibody from ZDR-5307, Beijing ZSGB Biotechnology were maintained at 37°C for 60 min.
    anti-mouse IgG
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Vero cells were applied to the reproduction of SARS-CoV-2 stocks.
    Vero
    suggested: CLS Cat# 605372/p622_VERO, RRID:CVCL_0059)
    Software and Algorithms
    SentencesResources
    Statistical analysis: All data were analyzed by GraphPad Prism 8.0 software.
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)

    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: 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 found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).

    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.

  3. Version published to 10.1101/2020.03.13.990036v2 on bioRxiv
  4. Version published to 10.1101/2020.03.13.990036v1 on bioRxiv