Immunogenic potential of DNA vaccine candidate, ZyCoV-D against SARS-CoV-2 in animal models

  1. Ayan Dey
  2. T.M. Chozhavel Rajanathan
  3. Harish Chandra
  4. Hari P.R. Pericherla
  5. Sanjeev Kumar
  6. Huzaifa S. Choonia
  7. Mayank Bajpai
  8. Arun K. Singh
  9. Anuradha Sinha
  10. Gurwinder Saini
  11. Parth Dalal
  12. Sarosh Vandriwala
  13. Mohammed A. Raheem
  14. Rupesh D. Divate
  15. Neelam L. Navlani
  16. Vibhuti Sharma
  17. Aashini Parikh
  18. Siva Prasath
  19. M. Sankar Rao
  20. Kapil Maithal

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Abstract

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  1. Version published to 10.1016/j.vaccine.2021.05.098
  2. ScreenIT

    SciScore for 10.1101/2021.01.26.428240: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIACUC: Animal Immunization: The immunogenicity study for the ZyCoV-D vaccine was carried out in inbred BALB/C mouse, guinea pig, and New Zealand white rabbit model after having ethical approval from Institutional Animal Ethics Committee, CPCSEA Reg. No.: 335/PO/RcBi/S/01/CPCSEA, with IAEC approved application numbers: VAC/010/2020 and VAC/013/2020.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    After 72Hrs, plates were fixed with 1:1 acetone and methanol Anti-S1 rabbit polyclonal antibody (Novus) was added to each well and incubated for 1Hr followed by incubation with FITC labelled anti-rabbit antibody (Merck).
    Anti-S1
    suggested: None
    anti-rabbit
    suggested: None
    After that, plates were again washed thrice followed by incubation with 1:5,000 dilution of horse radish peroxidase (HRP) conjugated anti-guinea pig IgG secondary antibody (Sigma-Aldrich) or 1:2,000 dilution of HRP conjugated anti-mouse IgG secondary antibody (Sigma-Aldrich) or 1:5,000 dilution of HRP conjugated anti rabbit IgG secondary antibody (Sigma-Aldrich) for 1 Hr at 37 °C.
    anti-guinea pig IgG
    suggested: None
    anti-mouse IgG
    suggested: None
    anti rabbit IgG
    suggested: None
    The 96 well Mouse IFN-γ ELISPOT kit (CTL, USA) plates pre-coated with purified anti-mouse IFN-γ capture antibody were taken out blocked with RPMI + 10% FBS +1X antibiotic for 1 Hr in CO2 incubator.
    anti-mouse IFN-γ
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    The virus was obtained from the BEI resources, USA (Isolate USA-WA1/2020), passaged and titrated in Vero-E6 cells.
    Vero-E6
    suggested: None
    Experimental Models: Organisms/Strains
    SentencesResources
    BALB/c mouse (five to seven-week-old), guinea pigs (five to seven-week-old) and New Zealand White rabbits (six to twelve-week-old) were used in this study.
    BALB/c
    suggested: RRID:IMSR_ORNL:BALB/cRl)
    Total DNA was extracted from Wistar rat tissues samples using the DNeasy Blood & Tissue Kit (Qiagen, Germany) as per manufacturer’s instructions.
    Wistar
    suggested: RRID:MGI:5657554)
    Software and Algorithms
    SentencesResources
    Inhibition concentration (IC50) of sera sample was calculated by plotting the percentage competition value obtained for each dilution verses serum dilution in a non-linear regression curve fit using Graph pad Prism 8.0.1 software.
    Graph pad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)
    The plates were dried and the spots were counted on ELISPOT Reader S6 Versa, (CTL USA) and analysed with Immunospot software version 7.0.
    Immunospot
    suggested: (ImmunoSpot Software for Analyzing ELISPOT Assays, RRID:SCR_011082)
    Statistical analysis: Statistical analysis of the results and graph creation were done with the Graph Pad Prism (version 8.0.1) and Microsoft Excel (version 7.0) for general statistical calculations, such as arithmetic mean and standard deviation.
    Microsoft Excel
    suggested: (Microsoft Excel, RRID:SCR_016137)

    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 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.
    • No funding statement was detected.
    • 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 10.1101/2021.01.26.428240v1 on bioRxiv