Potent universal-coronavirus therapeutic activity mediated by direct respiratory administration of a Spike S2 domain-specific human neutralizing monoclonal antibody

  1. Michael S. Piepenbrink
  2. Jun-Gyu Park
  3. Ashlesha Desphande
  4. Andreas Loos
  5. Chengjin Ye
  6. Madhubanti Basu
  7. Sanghita Sarkar
  8. David Chauvin
  9. Jennifer Woo
  10. Philip Lovalenti
  11. Nathaniel B. Erdmann
  12. Paul A. Goepfert
  13. Vu L. Truong
  14. Richard A. Bowen
  15. Mark R. Walter
  16. Luis Martinez-Sobrido
  17. James J. Kobie

Reviewed by

Read the full article

Listed in

Log in to save this article

Abstract

Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) marks the third novel β-coronavirus to cause significant human mortality in the last two decades. Although vaccines are available, too few have been administered worldwide to keep the virus in check and to prevent mutations leading to immune escape. To determine if antibodies could be identified with universal coronavirus activity, plasma from convalescent subjects was screened for IgG against a stabilized pre-fusion SARS-CoV-2 spike S2 domain, which is highly conserved between human β-coronavirus. From these subjects, several S2-specific human monoclonal antibodies (hmAbs) were developed that neutralized SARS-CoV-2 with recognition of all variants of concern (VoC) tested (Beta, Gamma, Delta, Epsilon, and Omicron). The hmAb 1249A8 emerged as the most potent and broad hmAb, able to recognize all human β-coronavirus and neutralize SARS-CoV and MERS-CoV. 1249A8 demonstrated significant prophylactic activity in K18 hACE2 mice infected with SARS-CoV-2 lineage A and lineage B Beta, and Omicron VoC. 1249A8 delivered as a single 4 mg/kg intranasal (i.n.) dose to hamsters 12 hours following infection with SARS-CoV-2 Delta protected them from weight loss, with therapeutic activity further enhanced when combined with 1213H7, an S1-specific neutralizing hmAb. As little as 2 mg/kg of 1249A8 i.n. dose 12 hours following infection with SARS-CoV Urbani strain, protected hamsters from weight loss and significantly reduced upper and lower respiratory viral burden. These results indicate in vivo cooperativity between S1 and S2 specific neutralizing hmAbs and that potent universal coronavirus neutralizing mAbs with therapeutic potential can be induced in humans and can guide universal coronavirus vaccine development.

Article activity feed

  1. ScreenIT

    SciScore for 10.1101/2022.03.05.483133: (What is this?)

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

    Table 1: Rigor

    EthicsConsent: The subjects provided informed consent.
    IRB: All procedures and methods were approved by the Institutional Review Board for Human Use at the University of Alabama at Birmingham, and all experiments were performed in accordance with relevant guidelines and regulations
    IACUC: Experiments were approved by the Texas Biomedical Research Institute and Colorado State University Biosafety and Animal Care and Use (IACUC) committees
    Euthanasia Agents: For the body weight and survival studies, five-week-old female K18 hACE2 transgenic mice were infected intranasally with 105 PFU per animal following gaseous sedation in an isoflurane chamber.
    Sex as a biological variableFive-week-old female K18 hACE2 transgenic mice were purchased from The Jackson Laboratory and maintained in the animal facility at Texas Biomedical Research Institute under specific pathogen-free conditions and ABSL3 containment.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    At 24 h p.i., infected cells were fixed with 10% neutral formalin for 24 h and were immune-stained using the anti-NP monoclonal antibody 1C7C7 (39).
    anti-NP
    suggested: (Sigma-Aldrich Cat# ZMS1075, RRID:AB_2893327)
    Experimental Models: Cell Lines
    SentencesResources
    CV3-25, S2P6, and CC40.8 were previously described (17–19) and heavy and light chain variable regions synthesized by IDT based on reported sequences (GenBank: MW681575.1, GenBank: MW681603.1 and (19)) and cloned into IgG1 expression vector for production in HEK293T cell.
    HEK293T
    suggested: RRID:CVCL_HA71)
    Cells and Viruses: African green monkey kidney epithelial cells (Vero E6, CRL-1586) were obtained from the American Type Culture Collection. A Vero E6 cell line expressing human ACE2 and TMPRSS2 (Vero AT) was obtained from BEI Resources (NR-54970).
    E6
    suggested: None
    Briefly, confluent monolayers of Vero E6 cells were mock infected or infected with the indicated virus.
    Vero E6
    suggested: RRID:CVCL_XD71)
    VeroE6/TMPRSS2 cells were seeded at 2 × 104 cells/well in opaque plates (Greiner 655083).
    VeroE6/TMPRSS2
    suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)
    Then 250,000 THP-1 cells (human monocytic cell line obtained from NIH AIDS Reagent Program) were added to the cells and incubated for 3 h at 37°C.
    THP-1
    suggested: None
    Briefly, 10-fold serial dilutions of samples were prepared in BA1 medium with antibiotics, inoculated onto confluent monolayers of Vero cells in 6-well plates,
    Vero
    suggested: None
    Experimental Models: Organisms/Strains
    SentencesResources
    K18 hACE2 transgenic mice experiments: All animal protocols involving K18 hACE2 transgenic mice were approved by the Texas Biomedical Research Institute IACUC (1718MU)
    K18 hACE2
    suggested: RRID:IMSR_GPT:T037657)
    Software and Algorithms
    SentencesResources
    Immunoglobulin sequences were analyzed by IgBlast (www.ncbi.nlm.nih.gov/igblast) and IMGT/V-QUEST (http://www.imgt.org/IMGT_vquest/vquest) to determine which sequences should lead to productive immunoglobulin, to identify the germline V(D)J gene segments with the highest identity, and to scrutinize sequence properties.
    IgBlast
    suggested: (IgBLAST, RRID:SCR_002873)
    A non-linear regression curve fit analysis over the dilution curve can be performed using GraphPad Prism to calculate NT50.
    GraphPad Prism
    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: 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.

    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.

  2. Version published to 10.1101/2022.03.05.483133v1 on bioRxiv