Rapid Manufacturing of Highly Cytotoxic Clinical-Grade SARS-CoV-2-specific T Cell Products Covering SARS-CoV-2 and Its Variants for Adoptive T Cell Therapy

• DOI: 10.3389/fbioe.2022.867042
• Publication Date: 2022-04-11T10:28:36Z
• Authors: Agnes Bonifacius; Sabine Tischer-Zi...; Maria Michela San...; Philip Mausberg; Josephine Schenk; Stephanie Koch; Johanna Barnstorf...; Nina Gödecke; Jörg Martens; Lilia Goudeva; Murielle Verboom; Jana Wittig; Britta Maecker-Ko...; Herrad Baurmann; Caren Clark; Olaf Brauns; Martina Simon; Peter Lang; Oliver A. Cornely; Michael Hallek; Rainer Blasczyk; Dominic Seiferling; Philipp Köhler; Britta Eiz-Vesper
• Keywords: 2019-20 coronavirus outbreak

Objectives: Evaluation of the feasibility SARS-CoV-2-specific T cell manufacturing for adoptive transfer in COVID-19 patients at risk to develop severe disease. Methods: Antiviral cells were detected blood convalescent following stimulation with PepTivator SARS-CoV-2 Select using Interferon-gamma Enzyme-Linked Immunospot (IFN-γ ELISpot), Cell Analysis Kit (Whole Blood) and Cytokine Secretion Assay (CSA) characterized respect memory phenotype, activation state cytotoxic potential by multicolor flow cytometry, quantitative real-time PCR multiplex analyses. Clinical-grade products generated MACS GMP CliniMACS Prodigy Capture System (IFN-gamma) (CCS). Functionality enriched was investigated cytotoxicity assays analysis secreted molecules upon target recognition. Results: Donor screening via IFN-γ ELISpot allows pre-selection donors generation cells. reactive against could be magnetically from peripheral small-scale CSA resembling clinical-grade CCS process showed an activated phenotype. Four successfully sufficient numbers purities comparable those observed donor pretesting CSA. The shown capable replicate, specifically recognize kill vitro secrete viability, total CD3 + number, proliferative capacity remained stable throughout storage up 72 h after end leukapheresis. Conclusion: are functional, have target-specific potential. Their function phenotype remain several days enrichment. partially matched, viable human lymphocytes collected individuals may provide opportunity support immune system