Immunization with synthetic SARS-CoV-2 S glycoprotein virus-like particles protects macaques from infection

  1. Guidenn Sulbaran
  2. Pauline Maisonnasse
  3. Axelle Amen
  4. Gregory Effantin
  5. Delphine Guilligay
  6. Nathalie Dereuddre-Bosquet
  7. Judith A. Burger
  8. Meliawati Poniman
  9. Marloes Grobben
  10. Marlyse Buisson
  11. Sebastian Dergan Dylon
  12. Thibaut Naninck
  13. Julien Lemaître
  14. Wesley Gros
  15. Anne-Sophie Gallouët
  16. Romain Marlin
  17. Camille Bouillier
  18. Vanessa Contreras
  19. Francis Relouzat
  20. Daphna Fenel
  21. Michel Thepaut
  22. Isabelle Bally
  23. Nicole Thielens
  24. Franck Fieschi
  25. Guy Schoehn
  26. Sylvie van der Werf
  27. Marit J. van Gils
  28. Rogier W. Sanders
  29. Pascal Poignard
  30. Roger Le Grand
  31. Winfried Weissenhorn

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Abstract

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  1. Version published to 10.1016/j.xcrm.2022.100528
  2. ScreenIT

    SciScore for 10.1101/2021.07.26.453755: (What is this?)

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

    Table 1: Rigor

    EthicsIRB: The protocols were approved by the institutional ethical committee “Comité d’Ethique en Expérimentation Animale du Commissariat à l’Energie Atomique et aux Energies Alternatives” (CEtEA #44) under statement number A20-011.
    Sex as a biological variablenot detected.
    RandomizationAnimals and study design: Cynomolgus macaques were randomly assigned in two experimental groups.
    Blindingnot detected.
    Power Analysis10 μl of sample were loaded into the capillary and intrinsic fluorescence was measured at a ramp rate of 1°C/min with an excitation power of 30 %.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    A horseradish peroxidase (HRP) conjugated goat anti-monkey H+L antibody (Invitrogen #PA1-84631) was then added and incubated for 1h before excess Ab was washed out and HRP substrate added.
    anti-monkey H+L
    suggested: (Thermo Fisher Scientific Cat# PA1-84631, RRID:AB_933605)
    CD3 (APC-Cy7, SP34-2, BD), CD4 (BV510, L200, BD) and CD8 (PE-Vio770, BW135/80, Miltenyi Biotec) antibodies was used as lineage markers.
    CD4
    suggested: None
    CD8
    suggested: None
    PE-Vio770, BW135/80, Miltenyi Biotec
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    The SARS-CoV-2 S RBD domain (residues 319 to 541) was expressed in EXPI293 cells by transient transfection according to the manufacturer’s protocol (Thermo Fisher Scientific).
    EXPI293
    suggested: RRID:CVCL_D615)
    Viruses and cells: For the macaques studies, SARS-CoV-2 virus (hCoV-19/France/ lDF0372/2020 strain) was isolated by the National Reference Center for Respiratory Viruses (Institut Pasteur, Paris, France) as previously described (Lescure et al., 2020) and produced by two passages on Vero E6 cells in DMEM (Dulbecco’s Modified Eagles Medium) without FBS, supplemented with 1% P/S (penicillin at 10,000 U ml−1 and streptomycin at 10,000 μg ml−1) and 1 μg ml−1 TPCK-trypsin at 37 °C in a humidified CO2 incubator and titrated on Vero E6 cells.
    Vero E6
    suggested: None
    Pseudovirus neutralization assay: Pseudovirus was produced by co-transfecting the pCR3 SARS-CoV-2-SΔ19 expression plasmid (Wuhan Hu-1; GenBank: MN908947.3) with the pHIV-1NL43 ΔEnv-NanoLuc reporter virus plasmid in HEK293T cells (ATCC, CRL-11268) (Caniels et al., 2021).
    HEK293T
    suggested: ATCC Cat# CRL-11268, RRID:CVCL_1926)
    HEK293T/ACE2 cells kindly provided by Dr. Paul Bieniasz (Schmidt et al., 2020) were seeded at a density of 20,000 cells/well in a 96-well plate coated with 50 μg/mL poly-L-lysine 1 day prior to the start of the neutralization assay.
    HEK293T/ACE2
    suggested: None
    Recombinant DNA
    SentencesResources
    For RBD expression, the following reagent was produced under HHSN272201400008C and obtained through BEI Resources, NIAID, NIH: Vector pCAGGS containing the SARS-Related Coronavirus 2, Wuhan-Hu-1 Spike Glycoprotein Receptor Binding Domain (RBD), NR-52309.
    pCAGGS
    suggested: RRID:Addgene_18926)
    Pseudovirus neutralization assay: Pseudovirus was produced by co-transfecting the pCR3 SARS-CoV-2-SΔ19 expression plasmid (Wuhan Hu-1; GenBank: MN908947.3) with the pHIV-1NL43 ΔEnv-NanoLuc reporter virus plasmid in HEK293T cells (ATCC, CRL-11268) (Caniels et al., 2021).
    pHIV-1NL43 ΔEnv-NanoLuc
    suggested: None
    The pCR3 SARS-CoV-2-SΔ19 expression plasmid contained the following mutations compared to the WT for the variants of concern: deletion (Δ) of H69, V70 and Y144, N501Y, A570D, D614G, P681H, T716I, S982A and D1118H in B.1.1.7; L18F, D80A, D215G, L242H, R246I, K417N, E484K, N501Y, D614G and A701V in B.1.351; L18F, T20N, P26S, D138Y, R190S, K417T, E484K, N501Y, D614G, H655Y and T1027I in P.1 (Caniels et al., 2021).
    pCR3 SARS-CoV-2-SΔ19
    suggested: None
    Software and Algorithms
    SentencesResources
    Antibody titers are presented as ED50 using the GraphPad Prism software version 6.
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)
    One million of PBMC were cultured in complete medium (RPMI1640 Glutamax+, Gibco; supplemented with 10 % FBS), supplemented with co-stimulatory antibodies (FastImmune CD28/CD49d, Becton Dickinson).
    Gibco
    suggested: None
    Next, cells were washed, stained with a viability dye (LIVE/DEAD fixable Blue dead cell stain kit, ThermoFisher), and then fixed and permeabilized with the BD Cytofix/Cytoperm reagent.
    ThermoFisher
    suggested: (ThermoFisher; SL 8; Centrifuge, RRID:SCR_020809)
    Analyses were performed with the FlowJo v.
    FlowJo
    suggested: (FlowJo, RRID:SCR_008520)

    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.

    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 10.1101/2021.07.26.453755v1 on bioRxiv