Chu Y, Milner J, Lamb M, Maryamchik E, Rigot O, Ayello J, Harrison L, Shaw R, Behbehani GK, Mardis ER, Miller K, Venkata LPR, Chang H, Lee D, Rosenthal E, Kadauke S, Bunin N, Talano JA, Johnson B, Wang Y, Cairo MS
BACKGROUND : Coronavirus disease 2019 (COVID-19) is caused by the 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 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 : 93% of convalescent donor blood samples passed the screening criteria for clinical manufacture. Three validation runs resulted in enriched T-cells that were 79% ± 21% IFN-γ+ T-cells. SARS-CoV-2-vCTLs displayed a highly diverse TCR 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 REGISTRATIONS : NCT04896606, NCT03266627, NCT03266640, NCT03266653, NCT04197596.