SARS-CoV-2 specific memory T lymphocytes from COVID-19 convalescent donors: identification, biobanking and large-scale production for Adoptive Cell Therapy

  1. C Ferreras
  2. B Pascual-Miguel
  3. C Mestre-Durán
  4. A Navarro-Zapata
  5. L Clares-Villa
  6. C Martín-Cortázar
  7. R De Paz
  8. A Marcos
  9. JL Vicario
  10. A Balas
  11. F García-Sánchez
  12. C Eguizabal
  13. C Solano
  14. M Mora-Rillo
  15. B Soria
  16. A Pérez-Martínez

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Abstract

SARS-CoV-2 is causing a second outbreak so the hope for its complete eradication is far from happening. In the absence of effective vaccines, it is mandatory to find effective treatments with low adverse effects able to treat hospitalized patients with COVID-19 disease. In this work, we determined the existence of SARS-CoV-2 specific T cells within the CD45RA − T memory cells from the blood of convalescent donors. Memory T cells can respond quickly to the infection and provide long-term immune protection to reduce the severity of the COVID-19 symptoms. Also, CD45RA − memory T cells confer protection from other pathogens the donors encountered in their life. This is vital to clear other secondary infections usually developed in hospitalized COVID-19 patients. SARS-CoV-2 specific memory T cells were found within all the CD45RA − subsets CD3 + , CD4 + , CD8 + , and in the central memory and effector memory subpopulations. The procedure to obtain the cells is feasible, easy to implement for small scale manufacture, quick and cost-effective involving minimal manipulation, and without GMP condition requirements. This biobank of specific SARS-CoV-2 memory T cells would be immediately available ‘off-the-shelf’ to treat moderate/severe cases of COVID-19 increasing the therapeutic options available for these patients.

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  1. ScreenIT

    SciScore for 10.1101/2020.10.23.352294: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementConsent: All the participants gave written consent with approval by the Hospital Institution Review Board (IRB number: 254/20) Cell processing and detection of SASR-CoV-2 specific memory T cells by IFN-γ assay: Peripheral Blood Mononuclear Cells (PBMCs) from healthy donors and the convalescent donors were isolated from their peripheral blood by density gradient centrifugation using Ficoll-Paque (GE Healthcare, Illinois, Chicago, USA).
    IRB: The donor gave written informed consent by the Declaration of Helsinki protocol, and the study was performed according to the guidelines of the local ethics committee (IRB Number 5579).
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variableThe median age of the convalescent donors was 37 years old (range 23-41), three were females and three were males.

    Table 2: Resources

    Antibodies
    SentencesResources
    Two healthy donors were enrolled who have not been exposed to COVID-19 patients and were tested negative for anti-SARS-CoV-2 antibodies in June 2020.
    anti-SARS-CoV-2
    suggested: None
    After 5 hours of stimulation, the cells were labeled with the IFN-γ Catch Reagent (IFN-γ Secretion Assay-Detection Kit, human Miltenyi Biotec) containing bispecific antibodies for CD45 and IFN-γ which was secreted by the stimulated target cells.
    IFN-γ
    suggested: None
    Software and Algorithms
    SentencesResources
    The analysis was performed using FlowJo 10.7.1 (FlowJo LLC)
    FlowJo
    suggested: (FlowJo, RRID:SCR_008520)
    Primers marked at their 5’ end with 6-FAM fluorochrome, allowed denatured fragment size analysis by capillary electrophoresis (ABI3130 DNA-analyzer) and Genemapper software (Thermo Fisher Scientific, USA)
    Genemapper
    suggested: (GeneMapper , RRID:SCR_014290)
    Two-tailed Mann-Whitney non-parametric test was used for comparison means for non-paired samples using GraphPad Prism (version 8.0.0 for Windows, GraphPad Software, San Diego, California USA).
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)
    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: We found the following clinical trial numbers in your paper:

    IdentifierStatusTitle
    NCT04578210RecruitingSafety Infusion of NatuRal KillEr celLs or MEmory T Cells as…

    Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

    Results from JetFighter: Please consider improving the rainbow (“jet”) colormap(s) used on pages 20, 21 and 22. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.

    Results from rtransparent:
    • Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
    • No funding statement was detected.
    • 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.

  2. Version published to 10.1101/2020.10.23.352294 on bioRxiv