SARS-CoV-2 Accessory Protein ORF8 Decreases Antibody-Dependent Cellular Cytotoxicity

  1. Guillaume Beaudoin-Bussières
  2. Ariana Arduini
  3. Catherine Bourassa
  4. Halima Medjahed
  5. Gabrielle Gendron-Lepage
  6. Jonathan Richard
  7. Qinghua Pan
  8. Zhen Wang
  9. Chen Liang
  10. Andrés Finzi

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Abstract

Viruses use many different strategies to evade host immune responses. In the case of SARS-CoV-2, its Spike mutates rapidly to escape from neutralizing antibodies. In addition to this strategy, ORF8, a small accessory protein encoded by SARS-CoV-2, helps immune evasion by reducing the susceptibility of SARS-CoV-2-infected cells to the cytotoxic CD8+ T cell response. Interestingly, among all accessory proteins, ORF8 is rapidly evolving and a deletion in this protein has been linked to milder disease. Here, we studied the effect of ORF8 on peripheral blood mononuclear cells (PBMC). Specifically, we found that ORF8 can bind monocytes as well as NK cells. Strikingly, ORF8 binds CD16a (FcγRIIIA) with nanomolar affinity and decreases the overall level of CD16 at the surface of monocytes and, to a lesser extent, NK cells. This decrease significantly reduces the capacity of PBMCs and particularly monocytes to mediate antibody-dependent cellular cytotoxicity (ADCC). Overall, our data identifies a new immune-evasion activity used by SARS-CoV-2 to escape humoral responses.

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  1. Version published to 10.3390/v14061237
  2. ScreenIT

    SciScore for 10.1101/2022.03.30.486403: (What is this?)

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

    Table 1: Rigor

    EthicsConsent: Ethics statement: The study was conducted in accordance with the Declaration of Helsinki in terms of informed consent and approval by an appropriate institutional
    IRB: The protocol was approved by the Ethics Committee of CHUM (protocol #19.381, approved on 25 March 2020).
    Sex as a biological variablenot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    ORF8 protein was probed with sheep anti-ORF8 antibody (dilution 1:1000, MRC Pure Reagents, Cat. DA088) for 16 hours at 4oC, followed by incubation with donkey anti-sheep HRP (horseradish peroxidase)-conjugated secondary antibody (1:2000, Invitrogen, Cat. AP184P) for 1 hour at room temperature.
    anti-ORF8
    suggested: None
    anti-sheep HRP
    suggested: (Millipore Cat# AP184P, RRID:AB_92611)
    Then, the anti-CD14-V450 (dilution 1:100, BD, Cat. 561390), anti-CD16-PE-Cy7 (dilution 1:100, BD, Cat. 560716) and anti-CD56-PE (dilution 1:100, BD, Cat. 556647) antibodies were added for 1 hour incubation on ice.
    anti-CD14-V450
    suggested: None
    anti-CD16-PE-Cy7
    suggested: None
    anti-CD56-PE
    suggested: (Beckman Coulter Cat# A07788, RRID:AB_2636814)
    Experimental Models: Cell Lines
    SentencesResources
    ORF8 Production: Three million HEK293T cells were seeded on 100 mm petri dishes.
    HEK293T
    suggested: None
    Software and Algorithms
    SentencesResources
    The data were analyzed with FlowJo.
    FlowJo
    suggested: (FlowJo, RRID:SCR_008520)
    Statistical analyses: Statistics were analyzed using GraphPad Prism version 8.0.2 (GraphPad, San Diego, CA, (USA).
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)
    GraphPad
    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.

  3. Version published to 10.1101/2022.03.30.486403v1 on bioRxiv