Full Brain and Lung Prophylaxis against SARS-CoV-2 by Intranasal Lentiviral Vaccination in a New hACE2 Transgenic Mouse Model or Golden Hamsters

  1. Min-Wen Ku
  2. Pierre Authié
  3. Maryline Bourgine
  4. François Anna
  5. Amandine Noirat
  6. Fanny Moncoq
  7. Benjamin Vesin
  8. Fabien Nevo
  9. Jodie Lopez
  10. Philippe Souque
  11. Catherine Blanc
  12. Sébastien Chardenoux
  13. llta Lafosse
  14. David Hardy
  15. Kirill Nemirov
  16. Françoise Guinet
  17. Francina Langa Vives
  18. Laleh Majlessi
  19. Pierre Charneau

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Abstract

Non-integrative, non-cytopathic and non-inflammatory lentiviral vectors are particularly suitable for mucosal vaccination and recently emerge as a promising strategy to elicit sterilizing prophylaxis against SARS-CoV-2 in preclinical animal models. Here, we demonstrate that a single intranasal administration of a lentiviral vector encoding a prefusion form of SARS-CoV-2 spike glycoprotein induces full protection of respiratory tracts and totally avoids pulmonary inflammation in the susceptible hamster model. More importantly, we generated a new transgenic mouse strain, expressing the human Angiotensin Converting Enzyme 2, with unprecedent brain permissibility to SARS-CoV-2 replication and developing a lethal disease in <4 days post infection. Even though the neurotropism of SARS-CoV-2 is now well established, so far other vaccine strategies under development have not taken into the account the protection of central nervous system. Using our highly stringent transgenic model, we demonstrated that an intranasal booster immunization with the developed lentiviral vaccine candidate achieves full protection of both respiratory tracts and brain against SARS-CoV-2.

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  1. ScreenIT

    SciScore for 10.1101/2021.02.03.429211: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIACUC: Ethical Approval of Animal Studies: Experimentation on mice and hamsters was realized in accordance with the European and French guidelines (Directive 86/609/CEE and Decree 87-848 of 19 October 1987) subsequent to approval by the Institut Pasteur Safety, Animal Care and Use Committee, protocol agreement delivered by local ethical committee (CETEA #DAP20007, CETEA #DAP200058) and Ministry of High Education and Research APAFIS#24627-2020031117362508 v1, APAFIS#28755-2020122110238379 v1.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variableMice: Female C57BL/6JRj mice (Janvier, Le Genest Saint Isle, France) were used between the age of 7 and 12 wks.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Experimental Models: Cell Lines
    SentencesResources
    Mouse Transgenesis: The human K18 promoter (GenBank: AF179904.1 nucleotide 90 to 2579) was amplified by nested PCR from A549 cell lysate, as described previously (Chow et al., 1997; Koehler et al., 2000).
    A549
    suggested: None
    Experimental Models: Organisms/Strains
    SentencesResources
    Mice: Female C57BL/6JRj mice (Janvier, Le Genest Saint Isle, France) were used between the age of 7 and 12 wks.
    C57BL/6JRj
    suggested: None
    Transgenic B6.K18-hACE2IP-THV mice were generated and bred, as detailed below at the CIGM of Institut Pasteur.
    B6.K18-hACE2IP-THV
    suggested: None
    ILV of high titre (4.16 × 109 TU/ml) carrying K18-hACE2IP-THV was used in transgenesis by subzonal micro-injection under the pellucida of fertilized eggs, and transplantation into the pseudo-pregnant B6CBAF1 females.
    B6CBAF1
    suggested: RRID:MGI:5652451)
    Eight N0 hACE2+ males were crossed with female WT C57BL/6 mice.
    C57BL/6
    suggested: None
    SARS-CoV-2 inoculation: Hamsters or transgenic B6.K18-hACE2IP-THV or B6.K18-ACE22Prlmn/JAX were anesthetized by i.p. injection of Ketamine and Xylazine mixture, transferred into a biosafety cabinet 3 and inoculated i.n. with 0.3 × 105 TCID50 of the BetaCoV/France/IDF0372/2020 SARS-CoV-2 clinical isolate (Lescure et al., 2020).
    B6.K18-ACE22Prlmn/JAX
    suggested: None
    Software and Algorithms
    SentencesResources
    , FcγII/III receptor blocking anti-CD16/CD32 (BD Biosciences), APC-anti-CD45 (BD), PerCP-Cy5.5-anti-CD3 (eBioscience), FITC-anti-CD4 (BD Pharmingen), BV711-anti-CD8 (BD Horizon), BV605-anti-CD69 (Biolegend), PE-anti-CCR7 (eBioscience) and VioBlue-Anti-B220 (Miltenyi).
    BD Biosciences
    suggested: (BD Biosciences, RRID:SCR_013311)
    Samples were acquired in an Attune NxT cytometer (Invitrogen) and data analyzed by FlowJo software (Treestar, OR, USA).
    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 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.

    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 10.1101/2021.02.03.429211 on bioRxiv