Peripheral innate and adaptive immune cells during COVID ‐19: Functional neutrophils, pro‐inflammatory monocytes, and half‐dead lymphocytes

  1. Emel Ekşioğlu‐Demiralp
  2. Servet Alan
  3. Uluhan Sili
  4. Dilek Bakan
  5. İlhan Ocak
  6. Rayfe Yürekli
  7. Nadir Alpay
  8. Serpil Görçin
  9. Alaattin Yıldız

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Abstract

Background

A better understanding of innate and adaptive cells in COVID‐19 is necessary for the development of effective treatment methods and vaccines.

Methods

We studied phenotypic features of innate and adaptive immune cells, oxidative burst, phagocytosis, and apoptosis. One hundred and three patients with COVID‐19 were grouped according to their clinical features into the categories of mild (35%), moderate (40.8%), and severe (24.3%).

Results

Monocytes were CD16 + pro‐inflammatory monocytes and tended to shed their HLA‐DR, especially in severe cases ( p  < 0.01). Neutrophils were mature and functional, although a decline of their CD10 and CD16 was observed ( p  < 0.01). No defect was found in the reactive oxygen species production and their apoptosis. The percentage of natural killer cells was in the normal range, whereas the percentages of CD8 + NK and CD56 + T lymphocytes were found to be high ( p  < 0.01). Although the absolute numbers of all lymphocyte subsets were low and showed a tendency for a gradual decrease in accordance with the disease progression, the most decreased absolute number was that of B lymphocytes, followed by CD4 + T cells in the severe cases. The percentages of double‐negative T cells; HLA‐DR + CD3 + and CD28 − CD8 + subsets were found to be significantly increased. Importantly, we demonstrated the increased baseline activation of caspase‐3 and increased lymphocyte apoptosis.

Conclusion

We suggest that SARS‐CoV‐2 primarily affects the lymphocytes and not the innate cells. The increased baseline activation of Caspase‐3 could make the COVID‐19 lymphocytes more vulnerable to cell death. Therefore, this may interrupt the crosstalk between the adaptive and innate immune systems.

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  1. Version published to 10.1002/cyto.b.22042
  2. ScreenIT

    SciScore for 10.1101/2020.08.01.20166587: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIRB: The study was approved by both Ministry of Health, Scientific Research Platform (no:2020-04-30_11_31) and Marmara University Ethics Committee (no:08.05.2020/09.2020.541).
    Consent: Informed consent was obtained from the participants before the study.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablePatients: A total of 103 patients infected with the new Coronavirus-2019, who applied to Istanbul Memorial Şişli Hospital and Marmara University Pendik Training and Research Hospital with various symptoms and signs from sore throat to shortness of breath were included in the study [52 female, 51 male with the mean age of 53,9 ± 15 (25-88)].

    Table 2: Resources

    Antibodies
    SentencesResources
    The following fluorochrome labelled monoclonal antibodies (mAb) and isotype-matched controls were used for two-three color phenotypic analysis: anti-IgG1, anti-IgG2a, anti-CD45, anti-CD3; anti-CD4; anti-CD8; anti-CD16; anti-CD56; anti-CD19; anti-CD20, anti-HLA-DR; anti-CD10; anti-CD25; anti-CD28, anti-CD69, (Becton&Dickinson Inc, San Jose, CA, USA).
    anti-IgG1
    suggested: None
    anti-IgG2a
    suggested: None
    anti-CD45
    suggested: (BD Biosciences Cat# 340418, RRID:AB_400423)
    anti-CD3
    suggested: (RayBiotech Cat# DS-MB-02991, RRID:AB_1544189)
    anti-CD4
    suggested: None
    anti-CD8
    suggested: (Thermo Fisher Scientific Cat# MA1-42127, RRID:AB_2537279)
    anti-CD16
    suggested: (Thermo Fisher Scientific Cat# MA1-42217, RRID:AB_2537342)
    anti-CD56
    suggested: (BD Biosciences Cat# 340417, RRID:AB_400422)
    anti-CD19
    suggested: (Thermo Fisher Scientific Cat# MA1-42134, RRID:AB_2537286)
    anti-CD20
    suggested: None
    anti-HLA-DR
    suggested: None
    anti-CD10
    suggested: None
    anti-CD25
    suggested: None
    anti-CD28
    suggested: None
    anti-CD69
    suggested: None
    Software and Algorithms
    SentencesResources
    Then the cells were immediately acquired by flow cytometry (FACSCalibur, Becton Dickinson Inc, San Jose, CA, USA) using CellQuest software, where at least 40,000 cells were acquired.
    FACSCalibur
    suggested: None
    CellQuest
    suggested: (BD CellQuest Pro, RRID:SCR_014489)
    Statistical analysis: The statistical software package SPSS version 25.0 was used for all statistical analyses.
    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:
    Our limitation was that we could not have measured cytokine release from isolated cell populations from patients. This issue requires further functional studies. In our group of patients, we have found that the absolute number of B lymphocytes was extremely reduced. Obviously, we should not expect proper immunoglobulin secretion from plasma cells in COVID-19 with that low number of B lymphocytes. It has already been shown that there was no peripheral B cell memory in Severe Acute Respiratory syndrom in six years follow-up (17, 45). In our study, we considered that this decrease might have been related to plasma cell differentiation. However, we demonstrated that no plasma cells have found in the periphery. A second reason might be the migration of B lymphocytes to the inflammation site. But, histopathological studies did not support this hypothesis. In addition, there is no finding such as lymphadenopathy. Furthermore, it has been shown that there are no lymphocytes in lymph node and spleen (46) Another possibility is that, as a member of the adaptive immune system, B lymphocytes may be one of the targets that directly excluded by SARS-CoV-2. As a part of imbalanced immune behavior, imbalanced cytokine release caused by SARS-CoV-2 may not permit proper involvement of B lymphocytes. More studies on B lymphocytes are require. The significant increase in the percentages of CD3+ and CD8+ T lymphocytes with their normal ranged absolute numbers in the mild patients can be considere...

    Results from TrialIdentifier: No clinical trial numbers were referenced.

    Results from Barzooka: We found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).

    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 10.1101/2020.08.01.20166587 on medRxiv