Generation of glucocorticoid-resistant SARS-CoV-2 T cells for adoptive cell therapy

Adoptive cell therapy with virus-specific T cells has been used successfully to treat life-threatening viral infections, supporting application of this approach to coronavirus disease 2019 (COVID-19). We expand severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) T cells from the peripheral blood of COVID-19-recovered donors and non-exposed controls using different culture conditions. We observe that the choice of cytokines modulates the expansion, phenotype, and hierarchy of antigenic recognition by SARS-CoV-2 T cells. Culture with interleukin (IL)-2/4/7, but not under other cytokine-driven conditions, results in more than 1,000-fold expansion in SARS-CoV-2 T cells with a retained phenotype, function, and hierarchy of antigenic recognition compared with baseline (pre-expansion) samples. Expanded cytotoxic T lymphocytes (CTLs) are directed against structural SARS-CoV-2 proteins, including the receptor-binding domain of Spike. SARS-CoV-2 T cells cannot be expanded efficiently from the peripheral blood of non-exposed controls. Because corticosteroids are used for management of severe COVID-19, we propose an efficient strategy to inactivate the glucocorticoid receptor gene (NR3C1) in SARS-CoV-2 CTLs using CRISPR-Cas9 gene editing. [Display omitted] •Expansion of SARS-CoV-2 T cells from COVID19-recovered donors is feasible•The choice of cytokines modulates the phenotype and function of SARS-CoV-2 T cells•SARS-CoV-2 T cells are predominantly CD4+ and skewed to the Spike RBD region•NR3C1 KO renders SARS-CoV-2 T cells steroid resistant without functional alteration Basar et al. demonstrate that SARS-CoV-2 T cells can be efficiently generated from COVID19-recovered donors with specificity against multiple structural SARS-CoV-2 proteins, including the Spike protein. Moreover, these cells can be genetically modified to render them resistant to corticosteroids, making their application clinically feasible.