Pathogenic neutrophilia drives acute respiratory distress syndrome in severe COVID-19 patients

  1. Devon J. Eddins
  2. Junkai Yang
  3. Astrid Kosters
  4. Vincent D. Giacalone
  5. Ximo Pechuan
  6. Joshua D. Chandler
  7. Jinyoung Eum
  8. Benjamin R. Babcock
  9. Brian S. Dobosh
  10. Mindy R. Hernández
  11. Fathma Abdulkhader
  12. Genoah L. Collins
  13. Richard P. Ramonell
  14. Christine Moussion
  15. Darya Y. Orlova
  16. Ignacio Sanz
  17. F. Eun-Hyung Lee
  18. Rabindra M. Tirouvanziam
  19. Eliver E.B. Ghosn

Reviewed by

Read the full article See related articles

Listed in

Log in to save this article

Abstract

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and the ensuing COVID-19 pandemic have caused ∼40 million cases and over 648,000 deaths in the United States alone. Troubling disparities in COVID-19-associated mortality emerged early, with nearly 70% of deaths confined to Black/African-American (AA) patients in some areas, yet targeted studies within this demographic are scant. Multi-omics single-cell analyses of immune profiles from airways and matching blood samples of Black/AA patients revealed low viral load, yet pronounced and persistent pulmonary neutrophilia with advanced features of cytokine release syndrome and acute respiratory distress syndrome (ARDS), including exacerbated production of IL-8, IL-1β, IL-6, and CCL3/4 along with elevated levels of neutrophil elastase and myeloperoxidase. Circulating S100A12 + /IFITM2 + mature neutrophils are recruited via the IL-8/CXCR2 axis, which emerges as a potential therapeutic target to reduce pathogenic neutrophilia and constrain ARDS in severe COVID-19.

Graphical Abstract

The lung pathology due to severe COVID-19 is marked by a perpetual pathogenic neutrophilia, leading to acute respiratory distress syndrome (ARDS) even in the absence of viral burden. Circulating mature neutrophils are recruited to the airways via IL-8 (CXCL8)/CXCR2 chemotaxis. Recently migrated neutrophils further differentiate into a transcriptionally active and hyperinflammatory state, with an exacerbated expression of IL-8 ( CXCL8 ), IL-1β ( IL1B ), CCL3, CCL4 , neutrophil elastase (NE), and myeloperoxidase (MPO) activity. Airway neutrophils and recruited inflammatory monocytes further increase their production of IL-8 ( CXCL8 ), perpetuating lung neutrophilia in a feedforward loop. MdCs and T cells produce IL-1β and TNF, driving neutrophils reprogramming and survival.

Article activity feed

  1. Version published to 10.1101/2021.06.02.446468v2 on bioRxiv
  2. ScreenIT

    SciScore for 10.1101/2021.06.02.446468: (What is this?)

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

    Table 1: Rigor

    EthicsIRB: All studies were approved by the Emory Institutional Review Board (IRB) under protocol numbers IRB00058507, IRB00057983 and IRB00058271.
    Consent: Informed consent was obtained from the patients when they had decision making ability or from a legal authorized representative (LAR) if the patient was unable to provide consent.
    Field Sample Permit: All work with infectious virus and respiratory samples from COVID-19 patients was conducted inside a biosafety cabinet within the Emory Health and Safety Office (EHSO)- and United States Department of Agriculture (USDA)-approved BSL3 containment facility in the Health Sciences Research Building at Emory University following protocols approved by the Institutional Biosafety Committee (IBC) and Biosafety Officer.
    IACUC: All work with infectious virus and respiratory samples from COVID-19 patients was conducted inside a biosafety cabinet within the Emory Health and Safety Office (EHSO)- and United States Department of Agriculture (USDA)-approved BSL3 containment facility in the Health Sciences Research Building at Emory University following protocols approved by the Institutional Biosafety Committee (IBC) and Biosafety Officer.
    Sex as a biological variablenot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Single-cell surface antibody-derived tag (ADT), encapsulation, and library generation: Leukocytes from whole blood and ETA samples were incubated with oligo-conjugated Ig-A/D/G/M for 10 mins at 4°C followed by the addition of Human TruStain FcX™ (BioLegend) and 10 min incubation at RT.
    Single-cell surface antibody-derived tag ( ADT) , encapsulation
    suggested: None
    library generation: Leukocytes from whole blood and ETA samples
    suggested: None
    ADT reads were aligned to a feature reference file containing the antibody-specific barcode sequences.
    antibody-specific barcode sequences .
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Plaque assays: Vero E6 cells were seeded in a 6-well plates (Falcon) with 5 x 105 cells/well in 5% DMEM 24 h prior to infection and checked to verify ≥80% confluency. 10-fold dilutions of virus, respiratory secretions, and/or scRNA-seq emulsion in serum-free DMEM (200 μL) were incubated on Vero E6 monolayers for 1 h absorption at 37°C with rocking at 15 min intervals.
    Vero E6
    suggested: None
    Software and Algorithms
    SentencesResources
    Cells were washed in ~4 mL FACS buffer, then resuspended in 200-1000 μL for acquisition using BD FACSDiva™ Software on the Emory Pediatric/Winship Flow Cytometry Core BD FACSymphony™ A5.
    BD FACSDiva™
    suggested: (BD FACSDiva Software, RRID:SCR_001456)
    Data were analyzed with FlowJo™ v10.7 (FlowJo LLC)
    FlowJo™
    suggested: (FlowJo, RRID:SCR_008520)
    FlowJo
    suggested: (FlowJo, RRID:SCR_008520)
    To recover neutrophils, scRNA-seq UMI count matrices were imported to R 4.0.2, and data analyses were performed using Seurat v4.042.
    Seurat
    suggested: (SEURAT, RRID:SCR_007322)
    The R package Slingshot was used to infer the trajectories of neutrophils recruited to the lungs.
    Slingshot
    suggested: (Slingshot, RRID:SCR_017012)
    Statistical analyses: Statistical analyses were performed using GraphPad Prism9.
    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 10.1101/2021.06.02.446468v1 on bioRxiv