Distinct cellular immune profiles in the airways and blood of critically ill patients with COVID-19

  1. Anno Saris
  2. Tom D.Y. Reijnders
  3. Esther J. Nossent
  4. Alex R. Schuurman
  5. Jan Verhoeff
  6. Saskia D. van Asten
  7. Hetty J. Bontkes
  8. Siebe G. Blok
  9. Janwillem Duitman
  10. Harm Jan Bogaard
  11. Leo Heunks
  12. Rene Lutter
  13. Tom van der Poll
  14. Juan J. Garcia Vallejo
  15. on behalf of the ArtDECO consortium and the Amsterdam UMC COVID study group

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Abstract

Our understanding of the coronavirus disease-19 (COVID-19) immune response is almost exclusively derived from studies that examined blood. To gain insight in the pulmonary immune response we analysed BALF samples and paired blood samples from 17 severe COVID-19 patients. Macrophages and T cells were the most abundant cells in BALF. In the lungs, both CD4 and CD8 T cells were predominantly effector memory cells and expressed higher levels of the exhaustion marker PD-1 than in peripheral blood. Prolonged ICU stay associated with a reduced proportion of activated T cells in peripheral blood and even more so in BALF. T cell activation in blood, but not in BALF, was higher in fatal COVID-19 cases. Increased levels of inflammatory mediators were more pronounced in BALF than in plasma. In conclusion, the bronchoalveolar immune response in COVID-19 has a unique local profile that strongly differs from the immune profile in peripheral blood.

Summary

The bronchoalveolar immune response in severe COVID-19 strongly differs from the peripheral blood immune profile. Fatal COVID-19 associated with T cell activation blood, but not in BALF.

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  1. ScreenIT

    SciScore for 10.1101/2020.10.29.360586: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementConsent: Informed consent for the use of samples and data was deferred until discharge from the ICU.
    IRB: Study procedure was approved by the Review Committee Biobank of the Amsterdam UMC (2020-065).
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Next, 1 million cells (or as many as available in case of BALFMCs) were stained with live/dead stain, and subsequently monoclonal antibodies added sequentially: CCR7, ⍰δ T cell receptor, all non-brilliant (ultra) violet (BV/BUV) or non-brilliant blue (BB) labelled markers and finally all BV, BUV and BB labelled antibodies.
    CCR7
    suggested: None
    Anti-RBD and anti-NP IgG antibodies were measured in EDTA plasma samples at 100-1200 fold dilutions using ELISA as previously described.
    Anti-RBD
    suggested: None
    anti-NP IgG
    suggested: None
    11 Plates were coated with RBD or N protein and specific IgG antibodies were detected using anti-human IgG (MH16, Sanquin).
    specific IgG
    suggested: None
    anti-human IgG
    suggested: (Cell Sciences Cat# MON5009, RRID:AB_419547)
    MH16
    suggested: None
    Software and Algorithms
    SentencesResources
    Correlations are represented using hierarchical edge bundling plot generated using the circlize package in R as previously described.18.
    circlize
    suggested: (circlize, RRID:SCR_002141)
    Statistical analysis was performed in the R statistical framework (Version 4.0.1, Vienna, Austria) or Graphad Prism v7.01 (GraphPad Software, San Diego, California, USA).
    Graphad Prism
    suggested: None
    Graphical presentation was performed using Graphpad Prism v7.01 (GraphPad Software), Adobe Illustrator CC v22.1 (Adobe, San Jose, California, USA), and R (Version 4.0.1).
    Graphpad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)
    GraphPad
    suggested: (GraphPad Prism, RRID:SCR_002798)
    Adobe Illustrator
    suggested: (Adobe Illustrator, RRID:SCR_010279)

    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: We detected the following sentences addressing limitations in the study:
    The present study is limited by its relatively low sample size – due to a rapidly declining incidence of SARS-CoV-2 infections after governmental measures were introduced – and variation in time of sampling, and these limitations necessitate caution when drawing conclusions. In conclusion, immune composition in the lungs of COVID-19 patients admitted to the ICU was substantially different than in peripheral blood. BALF mainly comprised macrophages and T cells, with high percentages of inflammatory monocyte-like macrophages and ⍰δ T cells especially after prolonged ICU stay. Both CD4 and CD8 T cells expressed higher levels of PD-1 in BALF as compared with PBMCs. Surprisingly, total CD8 T cell activation in BALF was lower than in peripheral blood. Reduced CD4 and CD8 T cell activation associated with extended ICU stay, especially in BALF, while peripheral activation of T cells (CD4 EM3 and EM4 as well as CD8 TEMRA) associated with mortality.

    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 10.1101/2020.10.29.360586 on bioRxiv