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. 2023 Mar 28;227(6):788-799.
doi: 10.1093/infdis/jiac500.

Manufacture and Characterization of Good Manufacturing Practice-Compliant SARS-COV-2 Cytotoxic T Lymphocytes

Yaya Chu  1 , Jordan Milner  1 , Margaret Lamb  2   3 , Elena Maryamchik  4   5 , Olivia Rigot  1 , Janet Ayello  1 , Lauren Harrison  1 , Rosemarie Shaw  1 , Gregory K Behbehani  6 , Elaine R Mardis  3   7 , Katherine Miller  7 , Lakshmi Prakruthi Rao Venkata  7 , Hsiaochi Chang  6 , Dean Lee  2   3 , Elana Rosenthal  1 , Stephan Kadauke  4 , Nancy Bunin  5   8 , Julie-An Talano  9 , Bryon Johnson  10 , Yongping Wang  4   5 , Mitchell S Cairo  1   11   12   13
Affiliations

Manufacture and Characterization of Good Manufacturing Practice-Compliant SARS-COV-2 Cytotoxic T Lymphocytes

Yaya Chu et al. J Infect Dis. .

Abstract

Background: Coronavirus disease 2019 (COVID-19) is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). SARS-CoV-2 virus-specific cytotoxic T-cell lymphocytes (vCTLs) could provide a promising modality in COVID-19 treatment. We aimed to screen, manufacture, and characterize SARS-CoV-2-vCTLs generated from convalescent COVID-19 donors using the CliniMACS Cytokine Capture System (CCS).

Methods: Donor screening was done by stimulation of convalescent COVID-19 donor peripheral blood mononuclear cells with viral peptides and identification of interferonγ (IFN-γ)+ CD4 and CD8 T cells using flow cytometry. Clinical-grade SARS-CoV-2-vCTLs were manufactured using the CliniMACS CCS. The enriched SARS-CoV-2-vCTLs were characterized by T-cell receptor sequencing, mass cytometry, and transcriptome analysis.

Results: Of the convalescent donor blood samples, 93% passed the screening criteria for clinical manufacture. Three validation runs resulted in enriched T cells that were 79% (standard error of the mean 21%) IFN-γ+ T cells. SARS-CoV-2-vCTLs displayed a highly diverse T-cell receptor repertoire with enhancement of both memory CD8 and CD4 T cells, especially in CD8 TEM, CD4 TCM, and CD4 TEMRA cell subsets. SARS-CoV-2-vCTLs were polyfunctional with increased gene expression in T-cell function, interleukin, pathogen defense, and tumor necrosis factor superfamily pathways.

Conclusions: Highly functional SARS-CoV-2-vCTLs can be rapidly generated by direct cytokine enrichment (12 hours) from convalescent donors.

Clinical trials registration: NCT04896606.

Keywords: TCRB CDR3 repertoire; IFN-γ+ T cells; SARS-CoV-2; cytokine capture system; immune landscape; virus-specific cytotoxic T lymphocytes.

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Conflict of interest statement

Potential conflicts of interest. M. S. C. has served as a consultant for Jazz Pharmaceuticals, Omeros Pharmaceuticals, and Novartis Pharmaceuticals; Speakers Bureau for Jazz Pharmaceuticals, Servier Pharmaceuticals, Amgen, Inc, Sanofi, and Sobi; and Advisory Board for Astra Zeneca. B. J. and S. K. have received research support from Miltenyi Biotec. Y. W. reports the Children's Hospital of Philadelphia Cell and Gene Therapy Laboratory has received free SARS-COV2 peptide reagent for testing. All other authors report no potential conflicts. All authors have submitted the ICMJE Form for Disclosure of Potential Conflicts of Interest. Conflicts that the editors consider relevant to the content of the manuscript have been disclosed.

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