Omicron-specific mRNA vaccine elicits potent immune responses in mice, hamsters, and nonhuman primates

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Abstract

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  1. SciScore for 10.1101/2022.03.01.481391: (What is this?)

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

    Table 1: Rigor

    Ethicsnot detected.
    Sex as a biological variableMouse immunizations: Female BALB/c mice (8–12 weeks old) were randomly allocated to groups.
    RandomizationMouse immunizations: Female BALB/c mice (8–12 weeks old) were randomly allocated to groups.
    Blindingnot detected.
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    The cells were stained with an anti-rabbit fluorescent IgG secondary antibody, and nucleus DNA was stained with DAPI (Sigma-Aldrich).
    anti-rabbit fluorescent IgG
    suggested: None
    Cells were blocked by Fc-receptor blockade with anti-CD16/CD32 (BD Biosciences), and then stained for 30 minutes at 4℃ with the following antibody panel each diluted in PBS: APC/Cyanine7 anti-mouse CD45 antibody (Biolegend) or FITC anti-mouse CD45.2 antibody (Biolegend), APC anti-mouse CD4 antibody (Biolegend),
    anti-mouse CD45
    suggested: None
    anti-mouse CD45.2
    suggested: None
    anti-mouse CD4
    suggested: None
    anti-mouse CD19 antibody (Biolegend), PE anti-mouse CD8a antibody (Biolegend) or PE anti-mouse/human CD45R/B220 antibody (Biolegend), PerCP/Cyanine5.5
    CD19
    suggested: None
    anti-mouse CD8a
    suggested: None
    anti-mouse/human CD45R/B220
    suggested: None
    anti-mouse/human CD44 antibody (Biolegend)
    anti-mouse/human CD44
    suggested: None
    APC anti-mouse CD138 (Syndecan-1) antibody (Biolegend), Brilliant Violet 510™ anti-mouse CD62L antibody (Biolegend), FITC anti-mouse CD107a (LAMP-1) antibody (Biolegend),
    anti-mouse CD138
    suggested: None
    Syndecan-1
    suggested: None
    anti-mouse CD62L
    suggested: None
    anti-mouse CD107a
    suggested: None
    LAMP-1
    suggested: None
    Then samples were blocked by anti-CD16/CD32 blockade as above, stained for 30 minutes at 4℃ with the following antibody: APC/Cyanine7 anti-mouse CD45 antibody (Biolegend),
    anti-CD16/CD32
    suggested: None
    anti-mouse CD4 antibody (Biolegend),
    CD4
    suggested: None
    anti-mouse CD8a antibody (Biolegend).
    CD8a
    suggested: None
    Cells were washed and fixed and permeabilized using Foxp3/Transcription factor staining buffer set (Invitrogen), and stained intracellularly for 30 minutes in PBS with antibodies including: PE anti-mouse IL-4 antibody (Biolegend) or PE anti-mouse IFN-γ antibody (Biolegend)
    anti-mouse IL-4
    suggested: None
    , APC anti-mouse IL-2 antibody (Biolegend) and FITC anti-human/mouse Granzyme B recombinant antibody (Biolegend).
    anti-mouse IL-2
    suggested: None
    anti-human/mouse
    suggested: None
    After incubation at 37℃, 5% CO2 for 18 h, the plates were washed, and biotinylated anti-mouse IFN-γ, IL-2, or IL-4 antibody was added to each well, following incubation of detection second antibodies.
    anti-mouse IFN-γ
    suggested: None
    IL-2
    suggested: None
    IL-4
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Vaccine antigen detection by immunofluorescence: Transfected HEK293T cells were fixed in 4% paraformaldehyde (PFA) and permeabilized in PBS/0.1% Triton X-100.
    HEK293T
    suggested: None
    Briefly, virus samples were serially 10-fold diluted with DMEM with 2.5% FBS, and inoculated to Vero cells or Vero E6 seeded overnight at 1.5 × 105 /well in 24-well plates; after incubated at 37°C for 1 h, the inoculate was replaced with DMEM containing 2.5% FBS and 0.9% carboxymethyl-cellulose.
    Vero
    suggested: None
    Vero E6
    suggested: None
    Experimental Models: Organisms/Strains
    SentencesResources
    Mouse immunizations: Female BALB/c mice (8–12 weeks old) were randomly allocated to groups.
    BALB/c
    suggested: RRID:IMSR_ORNL:BALB/cRl)
    Software and Algorithms
    SentencesResources
    RNA integrity was assessed by microfluidic capillary electrophoresis (Fragment Analyzer systems 5200, Agilent), and the concentration, pH, residual DNA, proteins, and dsRNA impurities of the solution were determined.
    Agilent
    suggested: (Agilent Bravo NGS, RRID:SCR_019473)
    RLUs of sample wells were normalized with positive control wells, and NT50 was calculated as EC50 by a normalized four-parameter sigmoid curve fit with constrains of EC50 > 0 and hillslope > 0 in Prism 8.0 (GraphPad)
    Prism
    suggested: (PRISM, RRID:SCR_005375)
    GraphPad
    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 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.

    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.