Table 1. SARS-CoV-2 CTL manufacture and... | American Society of Hematology

Table 1.

SARS-CoV-2 CTL manufacture and final characteristics

CD3⁺ cells	CD3⁺CD8⁺ cells	CD3⁺CD4⁺ cells	Tetramer positive out of CD3⁺CD8⁺ cells	Tetramer positive out of total cells	Cytotoxicity∗	Naïve T-cell content	Monocyte content	Natural killer cell content	B-cell content
96.5%	87.6%	4%	76.7%	68.5%	82%	0%	0.2%	0.7%	0.2%

CD3⁺ cells	CD3⁺CD8⁺ cells	CD3⁺CD4⁺ cells	Tetramer positive out of CD3⁺CD8⁺ cells	Tetramer positive out of total cells	Cytotoxicity∗	Naïve T-cell content	Monocyte content	Natural killer cell content	B-cell content
96.5%	87.6%	4%	76.7%	68.5%	82%	0%	0.2%	0.7%	0.2%

Target cells were pulsed with the specific SARS-CoV-2 target peptides to allow for binding to HLA-A∗02:01 before performing the assay. The CTLs were generated toward 7 HLA-A∗02:01–restricted SARS-CoV-2 peptides (from spike, nucleocapsid, nonstructural proteins 3, 7, and 8, and 2 from ORF3a proteins) using 1 apheresis product from 1 healthy donor who had COVID-19 illness ∼1 year earlier. The apheresis product was separated into monocytes and lymphocytes via elutriation. Dendritic cells were generated from monocytes and pulsed with the SARS-CoV-2 peptides and used for the first ex vivo resensitization. Thereafter, T cells were serially restimulated with peptide pulsed monocytes and enriched based on adherence of the target T-cell population to the monocytes. The contribution of each peptide to overall cytotoxicity was tested using HLA-A∗02:01 tetramers. Characteristics of the final product are shown above.

∗

Cytotoxicity is shown at an effector-to-target cell ratio (effector cells = SARS-CoV-2 CTLs) of 10:1.

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