SARS-CoV-2-Specific Memory T Lymphocytes From COVID-19 Convalescent Donors: Identification, Biobanking, and Large-Scale Production for Adoptive Cell Therapy
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Abstract
Syndrome coronavirus 2 (SARS-CoV-2) pandemic is causing a second outbreak significantly delaying the hope for the virus’ complete eradication. In the absence of effective vaccines, we need effective treatments with low adverse effects that can treat hospitalized patients with COVID-19 disease. In this study, we determined the existence of SARS-CoV-2-specific T cells within CD45RA – memory T cells in the blood of convalescent donors. Memory T cells can respond quickly to infection and provide long-term immune protection to reduce the severity of COVID-19 symptoms. Also, CD45RA – memory T cells confer protection from other pathogens encountered by the donors throughout their life. It is of vital importance to resolve other secondary infections that usually develop in patients hospitalized with COVID-19. We found SARS-CoV-2-specific memory T cells in all of the CD45RA – subsets (CD3 + , CD4 + , and CD8 + ) and in the central memory and effector memory subpopulations. The procedure for obtaining these cells is feasible, easy to implement for small-scale manufacture, quick and cost-effective, involves minimal manipulation, and has no GMP 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, thereby increasing the therapeutic options available for these patients.
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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 Statement Consent: 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).Randomization not detected. Blinding not … 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 Statement Consent: 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).Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable The median age of the convalescent donors was 37 years old (range 23-41), three were females and three were males. Table 2: Resources
Antibodies Sentences Resources 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-2suggested: NoneAfter 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-γsuggested: NoneSoftware and Algorithms Sentences Resources The analysis was performed using FlowJo 10.7.1 (FlowJo LLC) FlowJosuggested: (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) Genemappersuggested: (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 Prismsuggested: (GraphPad Prism, RRID:SCR_002798)GraphPadsuggested: (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:
Identifier Status Title NCT04578210 Recruiting Safety 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.
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