Replication kinetic, cell tropism and associated immune responses in SARS-CoV-2 and H5N1 virus infected human iPSC derived neural models

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   ScreenIT

   Mar 24, 2021

   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 Statement	not detected.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	not detected.
   Cell Line Authentication	Contamination: The cells were routinely checked for the presence of mycoplasma.

   Table 2: Resources

   Experimental Models: Cell Lines
   Sentences	Resources
   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 …

   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 Statement	not detected.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	not detected.
   Cell Line Authentication	Contamination: The cells were routinely checked for the presence of mycoplasma.

   Table 2: Resources

   Experimental Models: Cell Lines
   Sentences	Resources
   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
   Sentences	Resources
   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

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   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).

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   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 ...

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   Results from TrialIdentifier: No clinical trial numbers were referenced.

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

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

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   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.

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2. 

   Version published to 10.1101/2021.03.15.435472 on bioRxiv

   Mar 16, 2021