Relative expression of proinflammatory molecules in COVID‐19 patients who manifested disease severities

  1. Shireen Nigar
  2. SM Tanjil Shah
  3. Md Ali Ahasan Setu
  4. Sourav Dutta Dip
  5. Habiba Ibnat
  6. M. Touhidul Islam
  7. Selina Akter
  8. Iqbal Kabir Jahid
  9. M. Anwar Hossain

Reviewed by

Read the full article

Listed in

Log in to save this article

Abstract

Aggressive immune response, due to overexpressed proinflammatory molecules, has been characterized in coronavirus disease 2019 (COVID‐19) patients. Some of those mediators have a dual and opposite role on immune systems at play behind differential disease severities. We investigated the expression of some cytokines and chemokines in COVID‐19 patients in Bangladesh. We diagnosed the patients by detecting severe acute respiratory syndrome coronavirus 2 RNA in nasal swab samples by the real‐time RT‐PCR method. Thirty adult patients were preselected based on their disease severities and grouped into mild, moderate, and severe cases. Nine healthy volunteers participated in this study as a control. Relative expression of nine cytokines/chemokine in total leukocytes was semi‐quantified in SYBRgreen‐based real‐time quantitative reverse‐transcriptase polymerase chain reaction. We performed statistical tests on transformed log data using SPSS 24.0. At the onset of symptoms (Day 1), angiotensin‐converting enzyme 2 (ACE2) ( p  < 0.05) and interleukin (IL)‐6 ( p  > 0.05) were upregulated in all COVID‐19 groups, although the expression levels did not significantly correlate with disease severities. However, expressions of IL‐6, monocyte chemotactic protein‐1, macrophage inflammatory protein‐1α, tumor necrosis factor‐α (TNF‐α), RANTES (regulated upon activation, normal T cell expressed and secreted), and ACE2, on Day 14, were positively correlated with disease severities. Relative viral load at Day 1 showed no significant correlation with cytokine expression but had a significant positive correlation with RANTES and ACE2 expression on Day 14 ( p  < 0.05). Male patients had a higher level of IL‐6 than female patients on Day 1 ( p  < 0.05). All COVID‐19 patients showed upregulated cytokines and chemokines on Day 14 compared to Day 1 except TNF‐α. Female patients had a higher expression of ACE2 and IL‐12 on Day 14. Upregulated cytokines/chemokines at the convalescent stage, especially IL‐6, may help in targeting anticytokine therapy in post‐COVID‐19 patients' management.

Article activity feed

  1. Version published to 10.1002/jmv.27112
  2. ScreenIT

    SciScore for 10.1101/2021.04.01.21254770: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIACUC: Ethical permission and Sample collection: The Ethical Review Committee of the Jashore University of Science and Technology approved this study (ERC no: ERC/FBST/JUST/2020-49).
    Consent: We informed individual participants (COVID-19 patients) or their family members about the study protocol and took verbal consent.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    We exclude a sample from one volunteer in the final study due to the appearance of antibodies against SARS-CoV-2.
    SARS-CoV-2
    suggested: None
    Software and Algorithms
    SentencesResources
    We performed statistical tests based on transformed log data using SPSS 24.0 for Windows (SPSS, Inc., Chicago, IL, USA) and GraphPad Prism 8.0 (GraphPad Software, USA) to construct all the figures logarithmic scales.
    SPSS
    suggested: (SPSS, RRID:SCR_002865)
    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: We detected the following sentences addressing limitations in the study:
    There were a few limitations along with the small sample size in our study. We could not collect repeated blood samples after 14 days from severe patients admitted to ICU. We also estimated the relative expression levels of cytokines and chemokines instead of serum cytokine levels. However, live vaccines such as the bacillus Calmette-Guerin (BCG) are known to induce trained immunity (enhanced innate immune response to subsequent infections). COVID-19 cases reported being lower in countries with universal BCG vaccination programs (such as Bangladesh, Nepal, Bhutan, Japan) compared to those without the programs (such as the USA, Spain, Canada, Italy) [43]. We hypothesized that BCG vaccination might induce an immune response and reduce SARS-CoV-2 viremia and the severity of COVID-19 infections [44], and rapid recovery of SARS-CoV-2 infection in the country.

    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

    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 10.1101/2021.04.01.21254770v1 on medRxiv