Immunogenicity and protective efficacy of an intranasal live-attenuated vaccine against SARS-CoV-2

  1. Jun-Gyu Park
  2. Fatai S. Oladunni
  3. Mohammed A. Rohaim
  4. Jayde Whittingham-Dowd
  5. James Tollitt
  6. Matthew D.J. Hodges
  7. Nadin Fathallah
  8. Muhsref Bakri Assas
  9. Wafaa Alhazmi
  10. Abdullah Almilaibary
  11. Munir Iqbal
  12. Pengxiang Chang
  13. Renee Escalona
  14. Vinay Shivanna
  15. Jordi B. Torrelles
  16. John J. Worthington
  17. Lucy H. Jackson-Jones
  18. Luis Martinez-Sobrido
  19. Muhammad Munir

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Abstract

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  1. Version published to 10.1016/j.isci.2021.102941
  2. ScreenIT

    SciScore for 10.1101/2021.01.08.425974: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIRB: The project licence was approved by both the Home Office and the local AWERB committee of Lancaster University.
    IACUC: Biosafety (BSC) and Institutional Animal Care and Use (IACUC) committees, protocols BSC20- and 1722 MA 3, respectively.
    Randomizationnot detected.
    BlindingSections were stained with H&E and evaluated under light microscopy in a blinded manner by a board-certified veterinary pathologist at Texas Biomedical Research Institute.
    Power Analysisnot detected.
    Sex as a biological variableAnimals: BALB/c female mice were purchased from Charles River, UK and housed under specific pathogen free conditions at Lancaster University.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Experimental Models: Cell Lines
    SentencesResources
    The rNDV-WT and rNDV-S viruses were propagated in chicken embryonated eggs quantified in Vero cells using procedures we described before33. 293T cells and Vero cells were maintained in Dulbecco’s Modified Eagle Medium (DMEM) with 10% FBS as described previously33.
    Vero
    suggested: None
    293T
    suggested: None
    SARS-CoV-2 microneutralization assay: The day before infection, Vero-E6 cells were seeded at 1×104 cells/well into 96-well plates and incubated at 37°C in a 5% CO2 incubator overnight.
    Vero-E6
    suggested: None
    Confluent monolayers of Vero E6 cells (96-plate format, 4 × 104cells/well, duplicates) were infected with 10-fold serial dilutions of supernatants obtained from the nasal turbinate or lung homogenates from SARS-CoV-2 infected golden Syrian hamsters.
    Vero E6
    suggested: None
    Experimental Models: Organisms/Strains
    SentencesResources
    Animals: BALB/c female mice were purchased from Charles River, UK and housed under specific pathogen free conditions at Lancaster University.
    BALB/c
    suggested: RRID:IMSR_ORNL:BALB/cRl)
    Software and Algorithms
    SentencesResources
    The half maximal neutralizing concentration (NC50) for sera was determined using 4-parameter nonlinear regression curve fit to raw infectivity data measured as relative light units, or as the percentage of infected cells (GraphPad Prism).
    GraphPad
    suggested: (GraphPad Prism, RRID:SCR_002798)
    Viral neutralization was evaluated and quantified using ELISPOT, and a sigmoidal dose-response, non-linear regression curve was generated using GraphPad Prism to calculate median neutralization titer (NT50) in each of the serum samples.
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)
    Then cells were washed 3X with PBS and stained with the Vectastain ABC kit and developed using the DAB Peroxidase Substrate kit (Vector Laboratory, Inc, CA, USA), as previously described 1.
    Vector Laboratory
    suggested: None
    All samples were analysed on a Beckman Cytoflex and analysed with FlowJo Software (TreeStar).
    FlowJo
    suggested: (FlowJo, RRID:SCR_008520)
    Images were used for macroscopic pathology scoring analysis by measuring the distributions of pathological lesions, including consolidation, congestion, and pneumonic lesions using NIH ImageJ software.
    ImageJ
    suggested: (ImageJ, RRID:SCR_003070)
    Samples were analysed using Fiji software.
    Fiji
    suggested: (Fiji, RRID:SCR_002285)

    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.

    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. ScreenIT

    SciScore for 10.1101/2021.01.08.425974: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementThe project licence was approved by both the Home Office and the local AWERB committee of Lancaster University.Randomizationnot detected.BlindingSections were stained with H&E and evaluated under light microscopy in a blinded manner by a board-certified veterinary pathologist at Texas Biomedical Research Institute.Power Analysisnot detected.Sex as a biological variableMETHODS Animals BALB/c female mice were purchased from Charles River, UK and housed under specific pathogen free conditions at Lancaster University.Cell Line Authenticationnot detected.

    Table 2: Resources

    Experimental Models: Cell Lines
    SentencesResources
    The rNDV-WT and rNDV-S viruses were propagated in chicken embryonated eggs quantified in Vero cells using procedures we described before33. 293T cells and Vero cells were maintained in Dulbecco's Modified Eagle Medium (DMEM) with 10% FBS as described previously33.
    Vero
    suggested: None
    Neutralization assays using pseudotyped viruses Derivatives of 293T cells expressing ACE2 receptors were generated by transducing 293T cells with ACE2 receptor expressing vector.
    293T
    suggested: None
    SARS-CoV-2 microneutralization assay The day before infection, Vero-E6 cells were seeded at 1×104 cells/well into 96-well plates and incubated at 37°C in a 5% CO2 incubator overnight.
    Vero-E6
    suggested: None
    Confluent monolayers of Vero E6 cells (96-well plate format, 4 x 104 cells/well, quadruplicates) were infected with the virus-serum mixture for 1 h 37oC.
    Vero E6
    suggested: None
    Experimental Models: Organisms/Strains
    SentencesResources
    METHODS Animals BALB/c female mice were purchased from Charles River, UK and housed under specific pathogen free conditions at Lancaster University.
    BALB/c
    suggested: RRID:IMSR_APB:4790)
    Software and Algorithms
    SentencesResources
    The half maximal neutralizing concentration (NC50) for sera was determined using 4parameter nonlinear regression curve fit to raw infectivity data measured as relative light units, or as the percentage of infected cells (GraphPad Prism).
    GraphPad
    suggested: (GraphPad Prism, RRID:SCR_002798)
    Viral neutralization was evaluated and quantified using ELISPOT, and a sigmoidal dose-response, non-linear regression curve was generated using GraphPad Prism to calculate median neutralization titer (NT50) in each of the serum samples.
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)
    Then cells were washed 3X with PBS and stained with the Vectastain ABC kit and developed using the DAB Peroxidase Substrate kit (Vector Laboratory, Inc, CA, USA), as previously described 1.
    Vector Laboratory
    suggested: None
    All samples were analysed on a Beckman Cytoflex and analysed with FlowJo Software (TreeStar).
    FlowJo
    suggested: (FlowJo, RRID:SCR_008520)
    Images were used for macroscopic pathology scoring analysis by measuring the distributions of pathological lesions, including consolidation, congestion, and pneumonic lesions using NIH ImageJ software.
    ImageJ
    suggested: (ImageJ, RRID:SCR_003070)
    Samples were analysed using Fiji software.
    Fiji
    suggested: (Fiji, RRID:SCR_002285)

    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.

    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.

  4. Version published to 10.1101/2021.01.08.425974v1 on bioRxiv