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In vitro functionality and endurance of GMP-compliant point-of-care BCMA.CAR-T cells at different timepoints of cryopreservation

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Item Type:Article
Title:In vitro functionality and endurance of GMP-compliant point-of-care BCMA.CAR-T cells at different timepoints of cryopreservation
Creators Name:Jiang, G. and Neuber, B. and Hückelhoven-Krauss, A. and Höpken, U. and Ding, Y. and Sedloev, D. and Wang, L. and Reichman, A. and Eberhardt, F. and Wermke, M. and Rehm, A. and Müller-Tidow, C. and Schmitt, A. and Schmitt, M.
Abstract:The search for target antigens for CAR-T cell therapy against multiple myeloma defined the B-cell maturation antigen (BCMA) as an interesting candidate. Several studies with BCMA-directed CAR-T cell therapy showed promising results. Second-generation point-of-care BCMA.CAR-T cells were manufactured to be of a GMP (good manufacturing practice) standard using the CliniMACS Prodigy® device. Cytokine release in BCMA.CAR-T cells after stimulation with BCMA positive versus negative myeloma cell lines, U266/HL60, was assessed via intracellular staining and flow cytometry. The short-term cytotoxic potency of CAR-T cells was evaluated by chromium-51 release, while the long-term potency used co-culture (3 days/round) at effector/target cell ratios of 1:1 and 1:4. To evaluate the activation and exhaustion of CAR-T cells, exhaustion markers were assessed via flow cytometry. Stability was tested through a comparison of these evaluations at different timepoints: d0 as well as d + 14, d + 90 and d + 365 of cryopreservation. As results, (1) Killing efficiency of U266 cells correlated with the dose of CAR-T cells in a classical 4 h chromium-release assay. There was no significant difference after cryopreservation on different timepoints. (2) In terms of endurance of BCMA.CAR-T cell function, BCMA.CAR-T cells kept their ability to kill all tumor cells over six rounds of co-culture. (3) BCMA.CAR-T cells released high amounts of cytokines upon stimulation with tumor cells. There was no significant difference in cytokine release after cryopreservation. According to the results, BCMA.CAR-T cells manufactured under GMP conditions exerted robust and specific killing of target tumor cells with a high release of cytokines. Even after 1 year of cryopreservation, cytotoxic functions were maintained at the same level. This gives clinicians sufficient time to adjust the timepoint of BCMA.CAR-T cell application to the patient’s course of the underlying disease.
Keywords:Multiple Myeloma, CAR-T Cells, B-Cell Maturation Antigen (BCMA), Timepoint, Stability and Function of CAR-T Cells
Source:International Journal of Molecular Sciences
Page Range:1394
Date:23 January 2024
Official Publication:https://doi.org/10.3390/ijms25031394

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