Siglec-1 on dendritic cells mediates SARS-CoV-2 trans -infection of target cells while on macrophages triggers proinflammatory responses

  1. Daniel Perez-Zsolt
  2. Jordana Muñoz-Basagoiti
  3. Jordi Rodon
  4. Marc Elousa
  5. Dàlia Raïch-Regué
  6. Cristina Risco
  7. Martin Sachse
  8. Maria Pino
  9. Sanjeev Gumber
  10. Mirko Paiardini
  11. Jakub Chojnacki
  12. Itziar Erkizia
  13. Xabier Muñiz
  14. Ester Ballana
  15. Eva Riveira-Muñoz
  16. Marc Noguera
  17. Roger Paredes
  18. Benjamin Trinité
  19. Ferran Tarrés-Freixas
  20. Ignacio Blanco
  21. Victor Guallar
  22. Jorge Carrillo
  23. Julià Blanco
  24. Amalio Telenti
  25. Holger Heyn
  26. Joaquim Segalés
  27. Bonaventura Clotet
  28. Javier Martinez-Picado
  29. Júlia Vergara-Alert
  30. Nuria Izquierdo-Useros

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Abstract

COVID-19 pandemic is not yet under control by vaccination, and effective antivirals are critical for preparedness. Here we report that macrophages and dendritic cells, key antigen presenting myeloid cells (APCs), are largely resistant to SARS-CoV-2 infection. APCs effectively captured viruses within cellular compartments that lead to antigen degradation. Macrophages sense SARS-CoV-2 and released higher levels of cytokines, including those related to cytokine storm in severe COVID-19. The sialic acid-binding Ig-like lectin 1 (Siglec-1/CD169) present on APCs, which interacts with sialylated gangliosides on membranes of retroviruses or filoviruses, also binds SARS-CoV-2 via GM1. Blockage of Siglec-1 receptors by monoclonal antibodies reduces SARS-CoV-2 uptake and transfer to susceptible target cells. APCs expressing Siglec-1 and carrying SARS-CoV-2 are found in pulmonary tissues of non-human primates. Single cell analysis reveals the in vivo induction of cytokines in those macrophages. Targeting Siglec-1 could offer cross-protection against SARS-CoV-2 and other enveloped viruses that exploit APCs for viral dissemination, including those yet to come in future outbreaks.

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  1. ScreenIT

    SciScore for 10.1101/2021.05.11.443572: (What is this?)

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

    Table 1: Rigor

    EthicsIRB: Ethics statement: The institutional review board on biomedical research from Hospital Germans Trias i Pujol (HUGTiP) approved this study.
    Sex as a biological variablenot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    ACE2-mFc concentration was determined by sandwich ELISA using a goat anti-mouse IgG Fc (Jackson Immunoresearch, 115-006-071) for capture, a horseradish peroxidase (HRP) labeled F(ab)2 Goat anti-mouse IgG Fc (Jackson immunoresearch, 115-036-071) as secondary antibody, A purified mouse IgG (D50, NIH AIDS Reagent Program) as standard and o-phenylenediamine dihydrochloride (Sigma-Aldrich, #P8787-100TAB) as substrate.
    anti-mouse IgG
    suggested: (Jackson ImmunoResearch Labs Cat# 115-036-071, RRID:AB_2338524)
    The sections were incubated with SARS Nucleocapsid Protein Antibody (Rabbit polyclonal; Novus Biologicals, NB100-56576SS) and rabbit anti-human Anti-Sialoadhesin/CD169 antibody (clone SP216, Abcam, ab183356) for 1 h, followed by a double detection polymer system (Mach 2 Double Stain 2, Biocare Medical)
    SARS Nucleocapsid Protein Antibody ( Rabbit polyclonal; Novus Biologicals , NB100-56576SS )
    suggested: None
    NB100-56576SS
    suggested: None
    rabbit anti-human Anti-Sialoadhesin/CD169 antibody
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    HEK-293T (ATCC repository) were maintained in DMEM with 10% fetal bovine serum, 100 IU/mL penicillin and 100 μg/mL streptomycin (all from Invitrogen).
    HEK-293T
    suggested: None
    Raji B lymphocyte and Raji DC-SIGN cell lines (kindly provided by Y. Van Kooyke) were maintained in Roswell Park Memorial Institute medium (RPMI; Invitrogen) or RPMI plus 1 mg/mL geneticin (Invitrogen).
    Raji DC-SIGN
    suggested: None
    Pseudoviral fusion assay: Macrophages or DCs activated or not with IFNα as previously described along with HEK-293T ACE2 cells were exposed to equivalent MOI of VSVg or SARS-CoV-2 spike pseudotyped lentiviruses.
    HEK-293T ACE2
    suggested: None
    Cells not exposed to the virus were equally co-cultured with Vero E6 cells and used to set 100% of cell viability for each condition.
    Vero E6
    suggested: None
    Recombinant DNA
    SentencesResources
    HIV-1 reporter pseudoviruses expressing SARS-CoV-2 Spike protein and luciferase were generated using two plasmids. pNL4-3.Luc.R-.
    pNL4-3.Luc
    suggested: None
    SARS-CoV-2.SctΔ19 was generated (Geneart) from the full protein sequence of SARS-CoV-2 spike with a deletion of the last 19 amino acids in the C-terminal, human-codon optimized and inserted into pcDNA3.4-TOPO (Ou et al., 2020).
    pcDNA3.4-TOPO
    suggested: None
    VSV-G plasmid (kindly provided by A.
    VSV-G
    suggested: RRID:Addgene_138479)
    For protein production, Expi293F cells (Thermofisher Scientific) were transfected with ACE2-mFc vector at a density of 2.5 × 106 cells/mL using Expifectamine (
    ACE2-mFc
    suggested: None
    Software and Algorithms
    SentencesResources
    Samples were analyzed with FACS Canto (BD Biosciences) using FlowJo software to evaluate collected data.
    FlowJo
    suggested: (FlowJo, RRID:SCR_008520)
    Abberior STAR RED and Abberior STAR 580 signal was depleted with a donut-shaped 775-nm pulsed STED laser.
    STAR
    suggested: (STAR, RRID:SCR_004463)
    Acquired STED images were thresholded and filtered using Gaussian filter (Sigma (Radius) = 0.75) using Fiji (ImageJ distribution) software.
    Fiji
    suggested: (Fiji, RRID:SCR_002285)
    ImageJ
    suggested: (ImageJ, RRID:SCR_003070)
    The data originally used by Speranza et al. [10.1126/scitranslmed.abe8146] is publicly available in the Gene Expression Omnibus through GEO Series accession no. GSE156755 (www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE156755).
    Gene Expression Omnibus
    suggested: (Gene Expression Omnibus (GEO, RRID:SCR_005012)
    Due to the sparsity of the scRNAseq data we used MAGIC [https://doi.org/10.1016/j.cell.2018.05.061] denoising on the normalized count matrix to recover the underlying structure of the data and enhance the signal of lowly expressed genes.
    MAGIC
    suggested: (Magic, RRID:SCR_006406)
    Comparisons were performed with Graph Prism 9.
    Graph Prism
    suggested: None

    Results from OddPub: Thank you for sharing your data.

    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

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  2. Version published to 10.1101/2021.05.11.443572v1 on bioRxiv