Generation of Glucocorticoid Resistant SARS-CoV-2 T-Cells for Adoptive Cell Therapy

Adoptive cell therapy with viral-specific T cells has been successfully used to treat life-threatening viral infections, supporting the application of this approach against COVID-19. We expanded SARS-CoV-2 T-cells from the peripheral blood of COVID-19-recovered donors and non-exposed controls using different culture conditions. We observed that the choice of cytokines modulates the expansion, phenotype and hierarchy of antigenic recognition by SARS-CoV-2 T-cells. Culture with IL-2/4/7 but not other cytokine-driven conditions resulted in >1000 fold expansion in SARS-CoV-2 T-cells with a retained phenotype, function and hierarchy of antigenic recognition when compared to baseline (pre-expansion) samples. Expanded CTLs were directed against structural SARS-CoV-2 proteins, including the receptor-binding domain of Spike. SARS-CoV-2 T-cells could not be efficiently expanded from the peripheral blood of non-exposed controls. Since corticosteroids are used for the management of severe COVID-19, we developed an efficient strategy to inactivate the glucocorticoid receptor gene (NR3C1) in SARS-CoV-2 CTLs using CRISPR-Cas9 gene editing. Funding: Supported in part by the generous philanthropic contributions to The University of Texas MD Anderson Cancer Center AML Moonshot Program, by the grants from National Institute of Health, National Cancer Institute 5R01CA211044-04 and Cancer Center Support (CORE) Grant (CA016672) that support the Flow Cytometry and Cellular Imaging Facility and the RNA sequencing core facility at MD Anderson Cancer Center. Conflict of Interest: The authors declare no competing interests. Ethical Approval: Buffy coat units were processed from 500mL of whole blood collected from each of the 10 COVID-19 recovered donors (CoV-RD) and 20 mL of peripheral blood from 5 healthy donors were collected under local Institutional Review Board approved protocols (Lab02-0630 and PA13-0647) and following informed consent.