Modulation of immunosuppressant drug treatment to improve SARS-CoV-2 vaccine efficacy in mice

  1. Amy V. Paschall
  2. Ahmet Ozdilek
  3. Sydney L. Briner
  4. Melinda A. Brindley
  5. Fikri Y. Avci

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Abstract

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  1. Version published to 10.1016/j.vaccine.2021.12.058
  2. ScreenIT

    SciScore for 10.1101/2021.09.28.462156: (What is this?)

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

    Table 1: Rigor

    EthicsEuthanasia Agents: For tissue processing and subsequent flow cytometry, mice were euthanized through carbon dioxide in accordance with IACUC guidelines.
    IACUC: For tissue processing and subsequent flow cytometry, mice were euthanized through carbon dioxide in accordance with IACUC guidelines.
    Sex as a biological variableMice: Eight-week-old female BALB/c mice were obtained from Jackson Laboratories (Bar Harbor, ME) and housed at the University of Georgia.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    The antigenicity and the immunogenicity of the purified recombinant spike protein was validated (Supplementary Fig 1) against verified available recombinant spike protein (BEI Resources, NR52397) and anti-SARS-CoV-2 spike antibody clone A20085C (BioLegend).
    NR52397
    suggested: None
    anti-SARS-CoV-2
    suggested: (BioLegend Cat# 943201, RRID:AB_2888742)
    One 384-well plate was used to test one dilution and one antibody (IgG or IgM).
    IgM
    suggested: None
    Anti-IgG-AP (Southern BioTech 1030-04) and anti-IgM-AP (Southern BioTech 1020-04) were used to detect antibodies.
    Anti-IgG-AP
    suggested: None
    anti-IgM-AP
    suggested: None
    Cells were then stained with the following antibodies and stains (in multiple sets to prevent fluorophore overlap): CD4-FITC (BioLegend), CD8-PECy5 (BioLegend), CD3-PE(BioLegend), CD45-APCCy7 (BioLegend), CD19-PE (BioLegend)
    CD4-FITC
    suggested: None
    CD8-PECy5
    suggested: None
    CD45-APCCy7
    suggested: None
    CD19-PE
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    The spike protein was expressed in a serum-free medium by transient transfection of FreeStyle™ 293-F cells and purified by affinity chromatography using Nickel resins.
    293-F
    suggested: RRID:CVCL_6642)
    The virus-sera mixture was added to three replicate wells of Vero cells in a 96-well plate and incubated for 24 hr at 37°C.
    Vero
    suggested: None
    Experimental Models: Organisms/Strains
    SentencesResources
    Mice: Eight-week-old female BALB/c mice were obtained from Jackson Laboratories (Bar Harbor, ME) and housed at the University of Georgia.
    BALB/c
    suggested: RRID:IMSR_ORNL:BALB/cRl)
    Recombinant DNA
    SentencesResources
    Production of recombinant SARS-CoV-2 spike protein: The mammalian expression vector, pcDNA3.1+, with the codon-optimized nucleotide sequence of SARS-CoV-2 spike protein was a generous gift from Jarrod Mousa (University of Georgia).
    pcDNA3.1+
    suggested: RRID:Addgene_117272)
    Software and Algorithms
    SentencesResources
    Flow cytometry data was analyzed using FlowJo Single Cell Analysis Software (Treestar
    FlowJo
    suggested: (FlowJo, RRID:SCR_008520)
    Statistical Analysis: GraphPad Prism v8 was used for statistical analyses.
    Statistical Analysis: GraphPad Prism
    suggested: None
    GraphPad
    suggested: (GraphPad Prism, RRID:SCR_002798)
    Furthermore, effect sizes were calculated with No Drug (Fig1-2) or continuous drug treatment (Fig3-4) using Excel to find Cohen’s d (standardized mean difference).
    Excel
    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:
    One limitation of our study is the translation of mouse drug treatment and COVID-19 vaccination into human patients. We selected the drug treatment regimens for our mouse models based on previous research showing drug efficacy with no apparent effects on animal well-being. Also, we used intraperitoneal drug injections for optimal delivery and to minimize variations in individual drug deliveries. This method minimizes potential error in drug delivery, though one potential limit would be faster release into the circulatory system as opposed to subcutaneous or intramuscular injections as sometimes used in humans. Similarly, our generated spike protein was validated in both intraperitoneal and intramuscular injection models, and we used intraperitoneal vaccination in the discussed experiments to minimize the chance for error. While the purified spike protein has not as yet been used as a COVID-19 vaccine in the clinical practice, our validation (Supplementary Fig 2) in addition to the SARS-CoV-2 neutralization experiments (Fig2, Fig 4) show the relevance of this protein to established COVID-19 vaccine practices. Alum was used for spike protein vaccination to optimize immune response and antibody response. However, most currently approved COVID-19 vaccines use different formulations with non-alum adjuvants. While we show the overall importance of regulating immunosuppressant treatment regimens in COVID-19 vaccine strategies, the differences in drug delivery systems, as well as dos...

    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.

  3. Version published to 10.1101/2021.09.28.462156 on bioRxiv