Breadth of SARS-CoV-2 neutralization and protection induced by a nanoparticle vaccine
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Abstract
Coronavirus vaccines that are highly effective against current and anticipated SARS-CoV-2 variants are needed to control COVID-19. We previously reported a receptor-binding domain (RBD)-sortase A-conjugated ferritin nanoparticle (scNP) vaccine that induced neutralizing antibodies against SARS-CoV-2 and pre-emergent sarbecoviruses and protected non-human primates (NHPs) from SARS-CoV-2 WA-1 infection. Here, we find the RBD-scNP induced neutralizing antibodies in NHPs against pseudoviruses of SARS-CoV and SARS-CoV-2 variants including 614G, Beta, Delta, Omicron BA.1, BA.2, BA.2.12.1, and BA.4/BA.5, and a designed variant with escape mutations, PMS20. Adjuvant studies demonstrate variant neutralization titers are highest with 3M-052-aqueous formulation (AF). Immunization twice with RBD-scNPs protect NHPs from SARS-CoV-2 WA-1, Beta, and Delta variant challenge, and protect mice from challenges of SARS-CoV-2 Beta variant and two other heterologous sarbecoviruses. These results demonstrate the ability of RBD-scNPs to induce broad neutralization of SARS-CoV-2 variants and to protect animals from multiple different SARS-related viruses. Such a vaccine could provide broad immunity to SARS-CoV-2 variants.
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SciScore for 10.1101/2022.01.26.477915: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IACUC: Animals and immunizations: The study protocol and all veterinarian procedures were approved by the Bioqual IACUC per a memorandum of understanding with the Duke IACUC, and were performed based on standard operating procedures. Sex as a biological variable Male and female macaques per group were balanced when availability permitted. Randomization not detected. Blinding Samples were scored by a board-certified veterinary pathologist in a blinded manner. Power Analysis not detected. Cell Line Authentication Contamination: All cells were tested monthly for mycoplasma. Table 2: Resources
Antibodies Sentences Resources ACE2 and neutralizing antibody blocking assay: ELISA plates were coated as stated above … SciScore for 10.1101/2022.01.26.477915: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IACUC: Animals and immunizations: The study protocol and all veterinarian procedures were approved by the Bioqual IACUC per a memorandum of understanding with the Duke IACUC, and were performed based on standard operating procedures. Sex as a biological variable Male and female macaques per group were balanced when availability permitted. Randomization not detected. Blinding Samples were scored by a board-certified veterinary pathologist in a blinded manner. Power Analysis not detected. Cell Line Authentication Contamination: All cells were tested monthly for mycoplasma. Table 2: Resources
Antibodies Sentences Resources ACE2 and neutralizing antibody blocking assay: ELISA plates were coated as stated above with 2 μg/mL recombinant ACE-2 protein or neutralizing antibodies, then washed and blocked with 3% BSA in 1x PBS. ACE2suggested: NoneA peroxide block (Leica) was applied for 5 min to quench endogenous peroxidase activity prior to applying the SARS-CoV-2 antibody (1:2000, GeneTex, GTX135357). SARS-CoV-2suggested: (Leinco Technologies Cat# LT2000, RRID:AB_2893936)GTX135357suggested: (GeneTex Cat# GTX135357, RRID:AB_2868464)Experimental Models: Cell Lines Sentences Resources ACE2-Fc was expressed by transient transfection of Freestyle 293-F cells. 293-Fsuggested: RRID:CVCL_6642)Pseudovirions were produced in HEK 293T/17 cells (ATCC cat. no. CRL-11268) by transfection using Fugene 6 (Promega, Catalog #E2692). HEK 293T/17suggested: ATCC Cat# CRL-11268, RRID:CVCL_1926)Pseudovirions were produced by co-transfection of Lenti-X 293T cells with psPAX2(gag/pol), 293Tsuggested: KCB Cat# KCB 200744YJ, RRID:CVCL_0063)The supernatants were collected at 48 h after transfection and filtered through 0.45-μm membranes and titrated using HEK293T cells that express ACE2 and TMPRSS2 protein (293T-ACE2-TMPRSS2 cells). HEK293Tsuggested: NoneFor the neutralization assay, 50 μl of SARS-CoV-2 spike pseudovirions were pre-incubated with an equal volume of medium containing serum at varying dilutions at room temperature for 1 h, then virus-antibody mixtures were added to 293T-ACE2-TMPRSS2 cells in a 96-well plate. 293T-ACE2-TMPRSS2suggested: NoneBriefly, target cells were either Vero E6 cells after a 2 day-infection with SARS-CoV-2 USA-WA1/2020 or 293T cells 2-days post transfection with a SARS-CoV-2 S protein (D614) expression plasmid. Vero E6suggested: RRID:CVCL_A7UJ)Experimental Models: Organisms/Strains Sentences Resources Mouse immunization and challenge: Eleven-month-old female BALB/c mice were purchased from Envigo (#047) and were used for the SARS-CoV, SARS-CoV-2 WA-1, SARS-CoV-2 B.1.351, and RsSHC014-CoV protection experiments. BALB/csuggested: NoneRecombinant DNA Sentences Resources Pseudovirions for 293T/ACE2 infection were produced by co-transfection with a lentiviral backbone (pCMV ΔR8.2) and firefly luciferase reporter gene (pHR’ CMV Luc) (Naldini et al., 1996) pCMV ΔR8.2suggested: NonePseudovirions were produced by co-transfection of Lenti-X 293T cells with psPAX2(gag/pol), psPAX2suggested: RRID:Addgene_12260)The recombinant pcDNA3.1 plasmid was linearized, transcribed using MEGAscript T7 Transcription Kit (ThermoFisher, catalog # AM1334), and purified with MEGAclear Transcription Clean-Up Kit (ThermoFisher, catalog # AM1908). pcDNA3.1suggested: RRID:Addgene_79663)Software and Algorithms Sentences Resources All mutations were confirmed by full-length spike gene sequencing by Sanger Sequencing, using Sequencher and SnapGene for sequence analyses. Sequenchersuggested: (Sequencher, RRID:SCR_001528)SnapGenesuggested: (SnapGene, RRID:SCR_015052)Data analysis was performed using FlowJo 10 software (BD Biosciences). FlowJosuggested: (FlowJo, RRID:SCR_008520)Images were analyzed by 2D class averages using standard protocols with Relion 3.0 (Zivanov et al., 2018). Relionsuggested: (RELION, RRID:SCR_016274)Statistics Analysis: Data were plotted using Prism GraphPad 8.0. GraphPadsuggested: (GraphPad Prism, RRID:SCR_002798)Wilcoxon rank sum exact test was performed to compare differences between groups with p-value < 0.05 considered significant using SAS 9.4 (SAS Institute, Cary, NC). SAS Institutesuggested: (Statistical Analysis System, RRID:SCR_008567)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: We detected the following sentences addressing limitations in the study:Our study has several limitations. First, our study did not evaluate the durability of vaccine-induced immune responses and protection against SARS-CoV-2 variants. Second, we did not set up longer time intervals between the second and the third booster vaccination, to mimic 4-6 month boosting interval in humans. Lastly, we challenged the animals with WA-1 strain, the Beta variant and the Delta variant; future in vivo protection studies will be required upon availability of viral stocks of other SARS-CoV-2 variants such as the Omicron variant. Thus, our study demonstrates that scNP vaccines with SARS-CoV-2 spike or spike subunits confer potent protection in NHPs against WA-1, Beta and Delta variants, and that they induce neutralizing antibodies to all SARS-CoV-2 variants tested in vitro. These findings have important implications for development of the next generation of COVID-19 vaccines.
Results from TrialIdentifier: We found the following clinical trial numbers in your paper:
Identifier Status Title NCT04915768 Recruiting Evaluating the Safety and Immunogenicity of Stabilized CH505… NCT04177355 Recruiting Evaluating the Safety and Immunogenicity of HIV-1 BG505 SOSI… 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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