Newcastle disease virus (NDV) expressing the spike protein of SARS-CoV-2 as a live virus vaccine candidate

  1. Weina Sun
  2. Sarah R. Leist
  3. Stephen McCroskery
  4. Yonghong Liu
  5. Stefan Slamanig
  6. Justine Oliva
  7. Fatima Amanat
  8. Alexandra Schäfer
  9. Kenneth H. Dinnon
  10. Adolfo García-Sastre
  11. Florian Krammer
  12. Ralph S. Baric
  13. Peter Palese

Reviewed by

Read the full article See related articles

Listed in

Log in to save this article

Abstract

No abstract available

Article activity feed

  1. ScreenIT

    SciScore for 10.1101/2020.07.26.221861: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIACUC: Experiments were performed in accordance with protocols approved by the Icahn School of Medicine at Mount Sinai Institutional Animal Care and Use Committee (IACUC).
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variableMice immunizations: Ten-week old female BALB/cJ mice (Jackson Laboratories) were used.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    The blocking buffer was discarded and surface proteins were stained with anti-NDV rabbit serum or SARS-CoV-2 spike receptor-binding domain (RBD) specific human monoclonal antibody CR3022 (24, 25) for 2h at RT.
    anti-NDV
    suggested: None
    anti-NDV rabbit serum
    suggested: None
    specific human monoclonal antibody CR3022 ( 24 , 25 ) for 2h at RT
    suggested: None
    The primary antibodies were discarded, cells were then washed 3 times with PBS and incubated with goat anti-rabbit Alexa Fluor 488 or goat anti-human Alexa Fluor 488 secondary antibodies (Thermo Fisher Scientific) for 1h at RT.
    anti-rabbit
    suggested: None
    anti-human
    suggested: None
    To detect the spike protein of SARS-CoV-2, a mouse monoclonal antibody 2B3E5 kindly provided by Dr. Thomas Moran at ISMMS was used, while the HN protein was detected by a mouse monoclonal antibody 8H2 (MCA2822, Bio-Rad).
    MCA2822
    suggested: None
    ELISA plates were washed 3 times with PBST and 50 µL of sheep anti-mouse IgG-horseradish peroxidase (HRP) conjugated antibody (GE Healthcare) was added at a dilution of 1:3,000 in blocking solution.
    anti-mouse IgG-horseradish
    suggested: None
    Cells were then stained with 100 µL per well of a mouse monoclonal anti-NP antibody (1C7), kindly provided by Dr. Thomas Moran at ISMMS, at 1µg/ml for 1h at RT.
    anti-NP
    suggested: None
    Cells were washed with PBS and incubated with 100 µL per well Anti-mouse IgG HRP (Rockland) secondary antibody at 1:3,000 dilution in PBS containing 1% dry milk for 1h at RT.
    Anti-mouse IgG
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Vero E6 cells were also maintained in DMEM containing 10% FBS with 100 unit/ml P/S at 37 °C with 5% CO2.
    Vero E6
    suggested: None
    Briefly, Vero cells were seeded onto 96-well (Denville) tissue culture plates at 2.5 x 104 cells/well the day before infection.
    Vero
    suggested: None
    Experimental Models: Organisms/Strains
    SentencesResources
    Mice immunizations: Ten-week old female BALB/cJ mice (Jackson Laboratories) were used.
    BALB/cJ
    suggested: RRID:IMSR_JAX:000651)
    Software and Algorithms
    SentencesResources
    The plasmids were purified using QIAprep Spin Miniprep kit (Qiagen) for Sanger sequencing (Macrogen).
    Macrogen
    suggested: (Macrogen, RRID:SCR_014454)
    The sequences of the transgenes were confirmed by Sanger Sequencing (Genewiz)
    Genewiz
    suggested: (GENEWIZ, RRID:SCR_003177)
    The endpoint titers of serum IgG responses was graphed using GraphPad Prism 7.0.
    GraphPad Prism
    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 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.

    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.

  2. Version published to 10.1016/j.ebiom.2020.103132
  3. Version published to 10.1101/2020.07.26.221861v2 on bioRxiv
  4. ScreenIT

    SciScore for 10.1101/2020.07.26.221861: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementExperiments were performed in accordance with protocols approved by the Icahn School of Medicine at Mount Sinai Institutional Animal Care and Use Committee (IACUC).Randomizationnot detected.Blindingnot detected.Power Analysisnot detected.Sex as a biological variableMice immunizations Ten-week old female BALB/cJ mice (Jackson Laboratories) were used.Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    The blocking buffer was discarded and surface proteins were stained with anti-NDV rabbit serum or SARS-CoV-2 spike receptor-binding domain (RBD) specific human monoclonal antibody CR3022 (24, 25) for 2h at RT.
    anti-NDV rabbit serum
    suggested: None
          <div style="margin-bottom:8px">
        <div><b>specific human monoclonal antibody CR3022 ( 24 , 25 ) for 2h at RT</b></div>
        <div>suggested: None</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">The primary antibodies were discarded, cells were then washed 3 times with PBS and incubated with goat anti-rabbit Alexa Fluor 488 or goat antihuman Alexa Fluor 488 secondary antibodies (Thermo Fisher Scientific) for 1h at RT.</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>anti-rabbit</b></div>
        <div>suggested: None</div>
      </div>
    
      <div style="margin-bottom:8px">
        <div><b>antihuman Alexa Fluor 488</b></div>
        <div>suggested: None</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">To detect the spike protein of SARS- CoV-2, a mouse monoclonal antibody 2B3E5 kindly provided by Dr. Thomas Moran at ISMMS was used while the HN protein was detected by a mouse monoclonal antibody 8H2 (MCA2822, Bio-Rad).</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>MCA2822</b></div>
        <div>suggested: None</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">ELISA plates were washed 3 times with PBST and 50 µL of sheep anti-mouse IgG-horseradish peroxidase (HRP) conjugated antibody (GE Healthcare) was added at a dilution of 1:3,000 in blocking solution.</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>anti-mouse IgG-horseradish</b></div>
        <div>suggested: None</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">Cells were then stained with 100 µL per well of a mouse monoclonal anti-NP antibody (1C7), kindly provided by Dr. Thomas Moran at ISMMS, at 1µg/ml for 1h at RT.</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>anti-NP</b></div>
        <div>suggested: None</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">Cells were washed with PBS and incubated with 100 µL per well Anti- mouse IgG HRP (Rockland) secondary antibody at 1:3,000 dilution in PBS containing 1% dry milk for 1h at RT.</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>Anti-</b></div>
        <div>suggested: None</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">The surface of the cells was stained with anti-NDV rabbit serum or spike- specific monoclonal antibody CR3022 that recognizes the RBD.</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>anti-NDV</b></div>
        <div>suggested: None</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">Immunization of mice with NDV LaSota expressing the spike protein elicited potently binding and neutralizing antibodies To evaluate the immunogenicity of our NDV vectors expressing the S or S-F as vaccine candidates against SARS-CoV-2, a proof of principle study was performed in mice.</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>SARS-CoV-2</b></div>
        <div>suggested: None</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">The lungs of infected mice were fixed in 10% neutral buffered formalin for IHC staining using an anti-SARS-CoV-2 NP antibody.</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>anti-SARS-CoV-2 NP</b></div>
        <div>suggested: None</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">We also thank Dr. Thomas Moran for the 2B3E5 and 1C7 antibodies.</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>1C7</b></div>
        <div>suggested: None</div>
      </div>
    </td></tr><tr><td style="min-width:100px;text-align:center; padding-top:4px;" colspan="2"><b>Experimental Models: Cell Lines</b></td></tr><tr><td style="min-width:100px;text=align:center"><i>Sentences</i></td><td style="min-width:100px;text-align:center"><i>Resources</i></td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">Briefly, Vero cells were seeded onto 96-well (Denville) tissue culture plates at 2.5 x 104 cells/well the day before infection.</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>Vero</b></div>
        <div>suggested: None</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">The spike protein is incorporated into NDV particles To validate the expression of S and S-F as transgenes, Vero E6 cells were infected with WT NDV or NDV expressing the S or S-F.</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>Vero E6</b></div>
        <div>suggested: <a href="https://scicrunch.org/resources/Any/search?q=CVCL_XD71">CVCL_XD71</a></div>
      </div>
    </td></tr><tr><td style="min-width:100px;text-align:center; padding-top:4px;" colspan="2"><b>Software and Algorithms</b></td></tr><tr><td style="min-width:100px;text=align:center"><i>Sentences</i></td><td style="min-width:100px;text-align:center"><i>Resources</i></td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">The plasmids were purified using QIAprep Spin Miniprep kit (Qiagen) for Sanger sequencing (Macrogen).</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>Macrogen</b></div>
        <div>suggested: (Macrogen, <a href="https://scicrunch.org/resources/Any/search?q=SCR_014454">SCR_014454</a>)</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">The sequences of the transgenes were confirmed by Sanger Sequencing (Genewiz)</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>Genewiz</b></div>
        <div>suggested: (GENEWIZ, <a href="https://scicrunch.org/resources/Any/search?q=SCR_003177">SCR_003177</a>)</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">The endpoint titers of serum IgG responses was graphed using GraphPad Prism 7.0.</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>GraphPad Prism</b></div>
        <div>suggested: (GraphPad Prism, <a href="https://scicrunch.org/resources/Any/search?q=SCR_002798">SCR_002798</a>)</div>
      </div>
    </td></tr></table>

    Data from additional tools added to each annotation on a weekly basis.

    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 is not a substitute for expert review. SciScore checks for the presence and correctness of RRIDs (research resource identifiers) in the manuscript, and detects sentences that appear to be missing RRIDs. SciScore also checks to make sure that rigor criteria are addressed by authors. It does this by detecting sentences that discuss criteria such as blinding or power analysis. SciScore does not guarantee that the rigor criteria that it detects are appropriate for the particular study. Instead it assists authors, editors, and reviewers by drawing attention to sections of the manuscript that contain or should contain various rigor criteria and key resources. For details on the results shown here, including references cited, please follow this link.

  5. Version published to 10.1101/2020.07.26.221861v1 on bioRxiv