Replication Kinetics, Cell Tropism, and Associated Immune Responses in SARS-CoV-2- and H5N1 Virus-Infected Human Induced Pluripotent Stem Cell-Derived Neural Models

  1. Lisa Bauer
  2. Bas Lendemeijer
  3. Lonneke Leijten
  4. Carmen W. E. Embregts
  5. Barry Rockx
  6. Steven A. Kushner
  7. Femke M. S. de Vrij
  8. Debby van Riel

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Abstract

Infections with the recently emerged severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are often associated with neurological complications. Evidence suggests that SARS-CoV-2 enters the brain via the olfactory nerve; however, SARS-CoV-2 is only rarely detected in the central nervous system of COVID-19 patients.

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  1. Version published to 10.1128/msphere.00270-21
  2. ScreenIT

    SciScore for 10.1101/2021.03.15.435472: (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 variablenot detected.
    Cell Line AuthenticationContamination: The cells were routinely checked for the presence of mycoplasma.

    Table 2: Resources

    Experimental Models: Cell Lines
    SentencesResources
    Cell Lines: VeroE6 (ATCC® CRL 1586TM) cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM, Lonza, Breda, the Netherlands) supplemented with 10% fetal calf serum (FCS, Sigma-Aldrich, St. Louis, MO,
    VeroE6
    suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)
    Viruses: The SARS-CoV-2 isolate (isolate BetaCoV/Munich/BavPat1/2020; European Virus Archive Global #026V-03883; kindly provided by Dr. C. Drosten) was previously described by Lamers et al.51,52 The zoonotic HPAI H5N1 virus (A/Indonesia/5/2005) was isolated from a human patient and the virus was propagated once in embryonated chicken eggs and twice in MDCK cells.
    MDCK
    suggested: CLS Cat# 602280/p823_MDCK_(NBL-2, RRID:CVCL_0422)
    Experimental Models: Organisms/Strains
    SentencesResources
    WTC-11 Coriell #GM25256, obtained from the Gladstone Institute, San Francisco, USA] were differentiated to NPCs as previously described2 with slight modifications.
    WTC-11 Coriell #GM25256
    suggested: None

    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:
    However, one caveat of our study is that other cells such as microglia, oligodendrocytes and vascular cells (pericytes, endothelial cells) are not present. Therefore, we cannot exclude that SARS-CoV-2 can infect and possibly replicate efficiently in other cells of the CNS or neuronal cell types such as cortical PV interneurons, midbrain or hindbrain cell types. Despite the low proportion of SARS-CoV-2 infected cells and the fact that infection seemed to be abortive in the hiPSC derived neural cultures, we found evidence for cellular immune activation. In particular, SARS-CoV-2 infection of the neural cultures resulted in the induction of type-III-IFN and IFNλ2/3, but not type-I-IFN or type-II-IFN. This result is in accordance with earlier reports suggesting that SARS-CoV-2 triggers only very mild type-I and type-II-IFN responses, but does trigger a robust type-III-IFN response in cell culture, human airway epithelial cells, ferrets and SARS-CoV-2 infected individuals41,42. In addition, IL-8—a chemotactic factor that attracts leukocytes—was induced in all hiPSC-derived cultures. In lung tissue and peripheral venous blood serum of SARS-CoV-2 infected patients, elevated levels of IL-8 are associated with severe COVID-1943–45. Furthermore, IL-8 has been detected in the CSF of SARS-CoV-2 patients who developed encephalitis, which might be induced by the SARS-CoV-2 associated brain immune response, since SARS-CoV-2 RNA could not be detected in the CSF46. However, how exactly these ...

    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.

  3. Version published to 10.1101/2021.03.15.435472v1 on bioRxiv