Snake venom phospholipases A2 possess a strong virucidal activity against SARS-CoV-2 in vitro and block the cell fusion mediated by spike glycoprotein interaction with the ACE2 receptor

  1. Andrei E. Siniavin
  2. Maria A. Nikiforova
  3. Svetlana D. Grinkina
  4. Vladimir A. Gushchin
  5. Vladislav G. Starkov
  6. Alexey V. Osipov
  7. Victor I. Tsetlin
  8. Yuri N. Utkin

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Abstract

A new coronavirus was recently discovered and named severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In the absence of specific therapeutic and prophylactic agents, the virus has infected almost hundred million people, of whom nearly two million have died from the viral disease COVID-19. The ongoing COVID-19 pandemic is a global threat requiring new therapeutic strategies. Among them, antiviral studies based on natural molecules are a promising approach. The superfamily of phospholipases A2 (PLA2s) consists of a large number of members that catalyze the hydrolysis of phospholipids at a specific position. Here we show that secreted PLA2s from the venom of various snakes protect to varying degrees the Vero E6 cells widely used for the replication of viruses with evident cytopathic action, from SARS-CoV-2 infection PLA2s showed low cytotoxicity to Vero E6 cells and the high antiviral activity against SARS-CoV-2 with IC 50 values ranged from 0.06 to 7.71 μg/ml. Dimeric PLA2 HDP-2 from the viper Vipera nikolskii , as well as its catalytic and inhibitory subunits, had potent virucidal (neutralizing) activity against SARS-CoV-2. Inactivation of the enzymatic activity of the catalytic subunit of dimeric PLA2 led to a significant decrease in antiviral activity. In addition, dimeric PLA2 inhibited cell-cell fusion mediated by SARS-CoV-2 spike glycoprotein. These results suggest that snake PLA2s, in particular dimeric ones, are promising candidates for the development of antiviral drugs that target lipid bilayers of the viral envelope and may be good tools to study the interaction of viruses with host cell membranes.

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  1. ScreenIT

    SciScore for 10.1101/2021.01.12.426042: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board Statementnot detected.
    RandomizationThe percentage of cell-cell fusion was calculated by counting the fused cells in each well in five random fields using an Olympus (Japan) epifluorescence microscope.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line AuthenticationContamination: All cell lines were tested negative for mycoplasma contamination.

    Table 2: Resources

    Experimental Models: Cell Lines
    SentencesResources
    Cells, virus and plasmid: Vero E6 (ATCC CRL-1586) and 293T (ATCC CRL-3216) cells were grown in complete DMEM medium (Gibco, USA), supplemented with 10% fetal bovine serum (FBS; HyClone, USA), 1× L-Glutamine (PanEco, Russia) and 1× penicillin-streptomycin (PanEco, Russia).
    E6
    suggested: None
    293T
    suggested: None
    Then, the virus samples treated by PLA2 were diluted below IC50 and titrated on Vero E6 cells by limiting dilution assay (5-fold dilutions in six replicates) in 96-well plates.
    Vero E6
    suggested: None
    Software and Algorithms
    SentencesResources
    Dimeric phospholipases HDP-1 and HDP-2 were isolated from vipera V. nikolskii venom and separated into subunits HDP-1P (UniProtKB Q1RP79), HDP-2P (UniProtKB Q1RP78) and HDP-1I (UniProtKB A4VBF0) as described in 36.
    UniProtKB
    suggested: (UniProtKB, RRID:SCR_004426)
    Data were analyzed using GraphPad Prism 6 (GraphPad Software Inc.,
    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.

    About SciScore

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  2. Version published to 10.1101/2021.01.12.426042 on bioRxiv