Trivalent NDV-HXP-S Vaccine Protects against Phylogenetically Distant SARS-CoV-2 Variants of Concern in Mice
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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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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
Ethics IACUC: 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 variable Mouse immunization and challenge studies: Female BALB/c mice were used in all studies. Randomization not detected. Blinding All samples were analyzed in a blinded manner. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources The HN protein was detected by a mouse monoclonal antibody 8H2 (MCA2822, Bio-Rad). MCA2822suggested: NoneELISA plates were afterwards washed 3 times with PBST and 50 μL of anti-mouse … 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
Ethics IACUC: 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 variable Mouse immunization and challenge studies: Female BALB/c mice were used in all studies. Randomization not detected. Blinding All samples were analyzed in a blinded manner. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources The HN protein was detected by a mouse monoclonal antibody 8H2 (MCA2822, Bio-Rad). MCA2822suggested: NoneELISA 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-horseradishsuggested: NoneDuring this time the primary antibody was biotinylated according to manufacturer protocol (Thermo Scientific EZ-Link NHS-PEG4-Biotin) NHS-PEG4-Biotinsuggested: NoneBlocking 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-SARSsuggested: NoneThe 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 NPsuggested: NoneAn 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-mousesuggested: NoneExperimental Models: Cell Lines Sentences Resources VERO-TMPRSS2 cells (BPS Biosciences, #78081) were maintained in DMEM (Gibco) containing 10% (vol/vol) VERO-TMPRSS2suggested: JCRB Cat# JCRB1818, RRID:CVCL_YQ48)Specifically, transfected BSRT7 and DF-1 cells were washed with warm PBS and trypsinized. DF-1suggested: RRID:CVCL_XF08)BSRT7 cells were mixed with DF-1 cells (~1: 2.5) in a 10-cm dish. BSRT7suggested: 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-E6suggested: NoneExperimental Models: Organisms/Strains Sentences Resources Mouse immunization and challenge studies: Female BALB/c mice were used in all studies. BALB/csuggested: RRID:IMSR_ORNL:BALB/cRl)Recombinant DNA Sentences Resources 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/L289Asuggested: NoneThe 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-Ssuggested: NoneThe 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-NPsuggested: NonepTM1-Psuggested: NonepTM1-Lsuggested: NonepCI-T7optsuggested: NoneBriefly, 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: NoneSoftware and Algorithms Sentences Resources 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 Prismsuggested: (GraphPad Prism, RRID:SCR_002798)Statistical analyses were performed using Prism software (GraphPad). Prismsuggested: (PRISM, RRID:SCR_005375)GraphPadsuggested: (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:
Identifier Status Title NCT04871737 Active, not recruiting Study of a Live rNDV Based Vaccine Against COVID-19 NCT05181709 Recruiting A Live Recombinant Newcastle Disease Virus-vectored COVID-19… NCT04830800 Recruiting A Phase 1/2 Safety and Immunogenicity Trial of COVID-19 Vacc… NCT04764422 Recruiting Assess the Safety and Immunogenicity of NDV-HXP-S Vaccine in… NCT04993209 Recruiting Clinical 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.
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