1. SciScore for 10.1101/2021.07.16.452721: (What is this?)

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

    Table 1: Rigor

    EthicsIACUC: All experiments were performed in accordance with institutional guidelines from the Animal Research and Ethics Board.
    Euthanasia Agents: Bronchoalveolar lavage, lung, mediastinal lymph node, nasal turbinate and spleen mononuclear cell isolation: Mice were euthanized by exsanguination.
    Sex as a biological variablenot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    200 µg of Anti-CD4 (clone GK1.5) and anti-CD8 (clone 2.43) depleting, or an IgG isotype control antibodies (MilliporeSigma, Etobicoke, ON, Canada) were intraperitoneally administered as a single bolus 26-days post-vaccination.
    suggested: None
    suggested: None
    an IgG
    suggested: (Hangzhou HuaAn Biotechnology Cat# HA1006, RRID:AB_2819167)
    The membrane was incubated with primary antibodies (1:1000 Anti-VSV-G (ThermoFisher Scientific Waltham, MA, United States), 1:2000 Anti-NP (ThermoFisher Scientific Waltham, MA, United States) and 1:5000 GAPDH (MilliporeSigma, Etobicoke, ON, Canada) diluted in 5% skim milk, followed by anti-mouse and anti-rabbit IRDye secondary antibodies (LI-COR, Lincoln, NE, United States) diluted in 5 % skim milk.
    suggested: None
    suggested: None
    suggested: None
    suggested: None
    After washing, goat anti-mouse-biotin antibodies (Southern Biotech, Birmingham, AL, United States) IgA (1:2000), IgG (1:5000), IgG1 (1:5000), IgG2a (1:5000)) were diluted in reagent diluent and added to all wells.
    suggested: None
    suggested: None
    Experimental Models: Cell Lines
    Both trivalent vaccines were further amplified in HEK293 cells and subsequently purified by cesium chloride density gradient ultracentrifugation.
    suggested: CLS Cat# 300192/p777_HEK293, RRID:CVCL_0045)
    Transgene protein analysis by Western blot: A549 cells (CCL-185, ATCC, Manassas, VA, United States) were cultured at 37°C in DMEM supplemented with 10% FBS, 1 % HEPES pH 7.3, 1% L-glutamine and 100 U/mL of penicillin–streptomycin.
    suggested: None
    The Vero E6 culture media was then replaced with 100 µL of the serum-virus mixture and was incubated at 37°C for 72 hours.
    Vero E6
    suggested: None
    Experimental Models: Organisms/Strains
    Mice: Age-matched 6–8-wk-old wild-type female BALB/c, C57BL/6J, or B6.Cg-Tg (K18-ACE2) 2Prlmn/J mice were purchased from either Charles River Laboratories (Saint Constant, QC, Canada) or The Jackson Laboratory (Bar Harbor, ME, United States)
    suggested: None
    suggested: None
    B6.Cg-Tg ( K18-ACE2 ) 2Prlmn/J
    suggested: None
    SARS-CoV-2 strain SB3-TYAGNC was provided by Dr. Arinjay Banerjee, Dr. Karen Mossman, Dr. Samira Mubareka, and Dr. Rob Kozak and isolated as described previously (Banerjee et al., 2020).
    suggested: None
    Recombinant DNA
    This PCR product was cloned in pCY1 plasmid which contains the mCMV promoter.
    suggested: None
    Software and Algorithms
    Mice: Age-matched 6–8-wk-old wild-type female BALB/c, C57BL/6J, or B6.Cg-Tg (K18-ACE2) 2Prlmn/J mice were purchased from either Charles River Laboratories (Saint Constant, QC, Canada) or The Jackson Laboratory (Bar Harbor, ME, United States)
    Charles River Laboratories
    suggested: (Charles River Laboratories, RRID:SCR_003792)
    Stained cells were fixed and permeabilized with BD Cytofix/Cytoperm before incubation in BD Perm/Wash buffer (BD Biosciences, San Jose, CA, United States).
    BD Cytofix/Cytoperm
    suggested: None
    Data were analyzed using FlowJo software (version 10.1; Tree Star, Ashland, OR, United States).
    suggested: (FlowJo, RRID:SCR_008520)
    Tests were performed using GraphPad Prism software (Version 9
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)
    , Graphpad Software, La Jolla, CA, United States).
    suggested: (GraphPad, RRID:SCR_000306)

    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:
    These limitations of i.m. delivery, along with our current findings and those from others (Bricker et al., 2021; Hassan et al., 2020), should further bolster the global effort in developing respiratory mucosal-deliverable next-generation COVID-19 vaccines. In this regard, there have been at least two clinical trials testing either inhaled aerosol ChAdOx1nCov-19 (Singh et al., 2021) or intranasally delivered ChAd-SARS-CoV-2-S developed by scientists at WUSTL (Clinical Trial NCT04751682). Our current study has also provided the novel experimental evidence that both B and T cells are required for optimal protection. Of note, our data suggest that the clinical outcomes (body weight losses and mortality) may not always corroborate with viral burden, depending on both the immune pathway and relative potency of vaccine. We have observed that while Tri:HuAd-vaccinated B cell-deficient hosts well-protected in terms of clinical outcomes suffered impaired lung viral clearance, T cell-deficient hosts suffered both moderately worsened clinical outcomes and lung viral clearance. On the other hand, the robust i.n. Tri:ChAd vaccine strategy protected wildtype, B cell- and T cell-deficient hosts equally well in terms of clinical outcomes, whereas lack of B or T cells led to impaired lung viral clearance. Besides using T cell-depletion approach, the role of vaccine-induced T cell immunity in protection is supported further by our observations from B.1.135-infected K18-hACE2 animals immunized w...

    Results from TrialIdentifier: We found the following clinical trial numbers in your paper:

    NCT04751682Active, not recruitingSafety and Immunogenicity of an Intranasal SARS-CoV-2 Vaccin…

    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.

    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.

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