Trivalent NDV-HXP-S Vaccine Protects against Phylogenetically Distant SARS-CoV-2 Variants of Concern in Mice

  1. Irene González-Domínguez
  2. Jose Luis Martínez
  3. Stefan Slamanig
  4. Nicholas Lemus
  5. Yonghong Liu
  6. Tsoi Ying Lai
  7. Juan Manuel Carreño
  8. Gagandeep Singh
  9. Gagandeep Singh
  10. Michael Schotsaert
  11. Ignacio Mena
  12. Stephen McCroskery
  13. Lynda Coughlan
  14. Florian Krammer
  15. Adolfo García-Sastre
  16. Peter Palese
  17. Weina Sun

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Abstract

This manuscript describes an extended work on the Newcastle disease virus (NDV)-based vaccine focusing on multivalent formulations of NDV vectors expressing different prefusion-stabilized versions of the spike proteins of different SARS-CoV-2 variants of concern (VOC). We demonstrate here that this low-cost NDV platform can be easily adapted to construct vaccines against SARS-CoV-2 variants.

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  1. Version published to 10.1128/spectrum.01538-22
  2. ScreenIT

    SciScore for 10.1101/2022.03.21.485247: (What is this?)

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

    Table 1: Rigor

    EthicsIACUC: Animal experiments: All the animal experiments were performed in accordance with protocols approved by the Icahn School of Medicine at Mount Sinai (ISMMS) Institutional Animal Care and Use Committee (IACUC).
    Sex as a biological variableMouse immunization and challenge studies: Female BALB/c mice were used in all studies.
    Randomizationnot detected.
    BlindingAll samples were analyzed in a blinded manner.
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    The HN protein was detected by a mouse monoclonal antibody 8H2 (MCA2822, Bio-Rad).
    MCA2822
    suggested: None
    ELISA plates were afterwards washed 3 times with PBST and 50 μL of anti-mouse IgG-horseradish peroxidase (HRP) conjugated antibody (Cytiva, GE Healthcare) was added at a dilution of 1:3,000 in blocking solution.
    anti-mouse IgG-horseradish
    suggested: None
    During this time the primary antibody was biotinylated according to manufacturer protocol (Thermo Scientific EZ-Link NHS-PEG4-Biotin)
    NHS-PEG4-Biotin
    suggested: None
    Blocking solution was removed and 100 μl/well of biotinylated mAb 1C7C7, a mouse anti-SARS nucleoprotein monoclonal antibody generated at the Center for Therapeutic Antibody Development at The Icahn School of Medicine at Mount Sinai ISMMS
    anti-SARS
    suggested: None
    The plaques were immuno-stained with an anti-SARS-CoV-2 NP primary mouse monoclonal antibody 1C7C7 kindly provided by Dr. Thomas Moran at ISMMS.
    anti-SARS-CoV-2 NP
    suggested: None
    An HRP-conjugated goat anti-mouse secondary antibody was used at 1:2000 and the plaques were visualized using TrueBlue™ Peroxidase Substrate (SeraCare Life Sciences Inc.)
    anti-mouse
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    VERO-TMPRSS2 cells (BPS Biosciences, #78081) were maintained in DMEM (Gibco) containing 10% (vol/vol)
    VERO-TMPRSS2
    suggested: JCRB Cat# JCRB1818, RRID:CVCL_YQ48)
    Specifically, transfected BSRT7 and DF-1 cells were washed with warm PBS and trypsinized.
    DF-1
    suggested: RRID:CVCL_XF08)
    BSRT7 cells were mixed with DF-1 cells (~1: 2.5) in a 10-cm dish.
    BSRT7
    suggested: CCLV Cat# CCLV-RIE 0583, RRID:CVCL_RW96)
    Vero-E6 cells or Vero-TMPRSS2 were seeded onto 12-well plates in growth media at 1:5 and cultured for two days.
    Vero-E6
    suggested: None
    Experimental Models: Organisms/Strains
    SentencesResources
    Mouse immunization and challenge studies: Female BALB/c mice were used in all studies.
    BALB/c
    suggested: RRID:IMSR_ORNL:BALB/cRl)
    Recombinant DNA
    SentencesResources
    Briefly, the variant HXP-S were inserted into the pNDV_LS/L289A rescue plasmid (between P and M genes) by in-Fusion cloning (Clontech, CA, USA).
    pNDV_LS/L289A
    suggested: None
    The recombinant product was transformed into MAX Efficiency™ Stbl2™ Competent Cells (Thermo Fisher Scientific, MA, USA) to generate the pNDV-HXP-S rescue plasmid.
    pNDV-HXP-S
    suggested: None
    The next day, cells were transfected with 2 μg of pNDV-HXP-S, 1 μg of pTM1-NP, 0.5 μg of pTM1-P, 0.5 μg of pTM1-L and 1 μg of pCI-T7opt and were re-suspended in 250 μl of a modified Eagle’s Minimum Essential Medium (Opti-MEM; Gibco).
    pTM1-NP
    suggested: None
    pTM1-P
    suggested: None
    pTM1-L
    suggested: None
    pCI-T7opt
    suggested: None
    Briefly, the mammalian-cell codon-optimized nucleotide sequence of a soluble spike protein (amino acids 1-1,213) lacking the polybasic cleavage site, carrying two stabilizing mutations (K986P and V987P), a signal peptide, and at the C-terminus a thrombin cleavage site, a T4 fold-on trimerization domain, and a hexahistidine tag was cloned into the mammalian expression vector pCAGGS. https://www.beiresources.org/).
    pCAGGS.
    suggested: None
    Software and Algorithms
    SentencesResources
    Briefly, the mammalian-cell codon-optimized nucleotide sequence of a soluble spike protein (amino acids 1-1,213) lacking the polybasic cleavage site, carrying two stabilizing mutations (K986P and V987P), a signal peptide, and at the C-terminus a thrombin cleavage site, a T4 fold-on trimerization domain, and a hexahistidine tag was cloned into the mammalian expression vector pCAGGS. https://www.beiresources.org/).
    https://www.beiresources.org/
    suggested: (BEI Resource Repository, RRID:SCR_013698)
    Analysis was performed using GraphPad Prism 7 software.
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)
    Statistical analyses were performed using Prism software (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: We found the following clinical trial numbers in your paper:

    IdentifierStatusTitle
    NCT04871737Active, not recruitingStudy of a Live rNDV Based Vaccine Against COVID-19
    NCT05181709RecruitingA Live Recombinant Newcastle Disease Virus-vectored COVID-19…
    NCT04830800RecruitingA Phase 1/2 Safety and Immunogenicity Trial of COVID-19 Vacc…
    NCT04764422RecruitingAssess the Safety and Immunogenicity of NDV-HXP-S Vaccine in…
    NCT04993209RecruitingClinical Trial of the COVID-19 Vaccine (Recombinant, Inactiv…

    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 10.1101/2022.03.21.485247v1 on bioRxiv