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

• Published in: The Journal of infectious diseases 2023-03, Vol.227 (6), p.788-799
• Main Authors: Chu, Yaya, Milner, Jordan, Lamb, Margaret, Maryamchik, Elena, Rigot, Olivia, Ayello, Janet, Harrison, Lauren, Shaw, Rosemarie, Behbehani, Gregory K, Mardis, Elaine R, Miller, Katherine, Prakruthi Rao Venkata, Lakshmi, Chang, Hsiaochi, Lee, Dean, Rosenthal, Elana, Kadauke, Stephan, Bunin, Nancy, Talano, Julie-An, Johnson, Bryon, Wang, Yongping, Cairo, Mitchell S
• Format: Article
• Language: English
• Subjects: CD4 antigen; CD4-Positive T-Lymphocytes; CD8 antigen; CD8-Positive T-Lymphocytes; Clinical trials; Coronaviruses; COVID-19; COVID-19 Drug Treatment; Cytokines; Cytotoxicity; Flow cytometry; Gene expression; Humans; Immunological memory; Interferon-gamma; Leukocytes, Mononuclear; Lymphocytes; Lymphocytes T; Memory cells; Peripheral blood mononuclear cells; SARS-CoV-2; Severe acute respiratory syndrome coronavirus 2; T cell receptors; T-Lymphocytes, Cytotoxic; Transcriptomes; γ-Interferon
• ISSN: 0022-1899; 1537-6613
• DOI: 10.1093/infdis/jiac500

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. Highly functional SARS-CoV-2–vCTLs can be rapidly generated from convalescent COVID-19 donors using the CliniMACS Cytokine Capture System. SARS-CoV-2–vCTLs displayed a highly diverse TCR repertoire, enhanced memory CD8 and CD4 T cells, and increased gene expression in T-cell function and other pathways.