Longitudinal profiles of plasma gelsolin, cytokines and antibody expression predict COVID-19 severity and hospitalization outcomes

  1. Meshach Asare-Werehene
  2. Michaeline McGuinty
  3. Agatha Vranjkovic
  4. Yannick Galipeau
  5. Juthaporn Cowan
  6. Bill Cameron
  7. Curtis L. Cooper
  8. Marc-André Langlois
  9. Angela M. Crawley
  10. Benjamin K. Tsang

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Abstract

Background

Prognostic markers for COVID-19 disease outcome are currently lacking. Plasma gelsolin (pGSN) is an actin-binding protein and an innate immune marker involved in disease pathogenesis and viral infections. Here, we demonstrate the utility of pGSN as a prognostic marker for COVID-19 disease outcome; a test performance that is significantly improved when combined with cytokines and antibodies compared to other conventional markers such as CRP and ferritin.

Methods

Blood samples were longitudinally collected from hospitalized COVID-19 patients as well as COVID-19 negative controls and the levels of pGSN in μg/mL, cytokines and anti-SARS-CoV-2 spike protein antibodies assayed. Mean±SEM values were correlated with clinical parameters to develop a prognostic platform.

Results

pGSN levels were significantly reduced in COVID-19 patients compared to healthy individuals. Additionally, pGSN levels combined with plasma IL-6, IP-10 and M-CSF significantly distinguished COVID-19 patients from healthy individuals. While pGSN and anti-spike IgG titers together strongly predict COVID-19 severity and death, the combination of pGSN and IL-6 was a significant predictor of milder disease and favorable outcomes.

Conclusion

Taken together, these findings suggest that multi-parameter analysis of pGSN, cytokines and antibodies could predict COVID-19 hospitalization outcomes with greater certainty compared with conventional clinical laboratory markers such as CRP and ferritin. This research will inform and improve clinical management and health system interventions in response to SARS-CoV-2 infection.

Trial Registration

N/A

Funding

The Ottawa Hospital Department of Medicine - Special Pandemic Agile Research Competition

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  1. ScreenIT

    SciScore for 10.1101/2022.06.01.22275882: (What is this?)

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

    Table 1: Rigor

    EthicsIRB: Ethics statement: This study was approved by the Ottawa Health Science Network Research Ethics Board (OH SNREB; Protocol# 20200200-01H) and conducted in accordance with the appropriate guidelines.
    Consent: Written informed consent was obtained from all subjects.
    Sex as a biological variablenot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    The detection antibody was raised against human plasma (soluble) gelsolin.
    human plasma (soluble) gelsolin.
    suggested: None
    Anti-spike protein antibody assay: A manual colorimetric ELISA was used to measure antibodies targeting SARS-CoV-2 spike protein, a complete description of the methods can be found here (36).
    Anti-spike protein
    suggested: None
    Additionally, positive and negative serum controls alongside an isotype-antigen specific calibration curve (CR3022 Human IgG1 (Absolute Antibody, Ab01680-10.0), anti-SARS-CoV-2 S CR3022 Human IgA (Absolute Antibody, Ab01680-16.0), or anti-SARS-CoV-2 S CR3022 Human IgM (Absolute Antibody, Ab01680-15.0)) was added to the plate and incubated for 2h with shaking.
    Human IgG1
    suggested: (Imported from the IEDB Cat# CR3022, RRID:AB_2848080)
    anti-SARS-CoV-2 S CR3022 Human IgA
    suggested: None
    anti-SARS-CoV-2 S CR3022 Human IgM
    suggested: None
    The plates were washed again and 50uL of isotype specific secondary antibodies (anti-human IgG#5-HRP (NRC), anti-human IgA-HRP (Jackson ImmunoResearch Labs, 109-035-011), and anti-human IgM-HRP (Jackson ImmunoResearch Labs, 109-035-129) added and incubated with shaking for 1 hour.
    anti-human IgG#5-HRP
    suggested: None
    anti-human IgA-HRP
    suggested: (SouthernBiotech Cat# 2050-05, RRID:AB_2687526)
    anti-human IgM-HRP
    suggested: (MyBioSource Cat# MBS673990, RRID:AB_10891687)
    Software and Algorithms
    SentencesResources
    Cytokine Assay: The concentrations of pro- and anti-inflammatory cytokines in plasma were quantified using multiplexing immunobead assays analyzed using the BioRad Luminex machine (Bio-Rad Laboratories, Hercules, CA,
    Bio-Rad Laboratories
    suggested: (Bio-Rad Laboratories, RRID:SCR_008426)
    Biostatistical methods: The GraphPad Prism 8 (San Diego, CA, USA) and SPSS software version 28
    GraphPad
    suggested: (GraphPad Prism, RRID:SCR_002798)
    8 (SPSS Inc., Chicago, IL, USA) were used to perform all statistical analyses and two-sided P ≤ 0.05 considered to indicate statistical significance.
    SPSS
    suggested: (SPSS, RRID:SCR_002865)

    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:
    Despite these promising findings, there are limitations that need to be acknowledged. The sample size of this study is relatively small due to the difficulty in sampling patients longitudinally in a hospital setting. Antibody kinetics (levels and isotype) during the early infection could be influenced by time since infection. Also, the control samples consist of out-patient populations that had no recorded history of respiratory infection during sample collection. Having control samples from patients who are SARS-CoV-2 negative but positive for other respiratory diseases will further strengthen the reliability of pGSN multi-analyte panels. In future studies, patient sample size will be increased while we investigate the prognostic significance of pGSN multi-analyte panel in SARS-CoV-2 variant patients, vaccinated patients as well as convalescent plasma.

    Results from TrialIdentifier: We found the following clinical trial numbers in your paper:

    IdentifierStatusTitle
    NCT04358406Active, not recruitingRhu-pGSN for Severe Covid-19 Pneumonia

    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.
    • Thank you for including a protocol registration statement.

    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.

  2. Version published to 10.1101/2022.06.01.22275882v1 on medRxiv