In vitro functionality and endurance of GMP-compliant point-of-care BCMA.CAR-T cells at different timepoints of cryopreservation
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...
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| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article (Journal) |
| Language: | English |
| Published: |
23 January 2024
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| In: |
International journal of molecular sciences
Year: 2024, Volume: 25, Issue: 3, Pages: 1-15 |
| ISSN: | 1422-0067 |
| DOI: | 10.3390/ijms25031394 |
| Online Access: | Verlag, kostenfrei, Volltext: https://doi.org/10.3390/ijms25031394 Verlag, kostenfrei, Volltext: https://www.mdpi.com/1422-0067/25/3/1394 
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| Author Notes: | Genqiao Jiang, Brigitte Neuber, Angela Hückelhoven-Krauss, Uta E. Höpken, Yuntian Ding, David Sedloev, Lei Wang, Avinoam Reichman, Franziska Eberhardt, Martin Wermke, Armin Rehm, Carsten Müller-Tidow, Anita Schmitt and Michael Schmitt |
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| 245 | 1 | 0 | |a In vitro functionality and endurance of GMP-compliant point-of-care BCMA.CAR-T cells at different timepoints of cryopreservation |c Genqiao Jiang, Brigitte Neuber, Angela Hückelhoven-Krauss, Uta E. Höpken, Yuntian Ding, David Sedloev, Lei Wang, Avinoam Reichman, Franziska Eberhardt, Martin Wermke, Armin Rehm, Carsten Müller-Tidow, Anita Schmitt and Michael Schmitt |
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| 520 | |a 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. | ||
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| 650 | 4 | |a CAR-T cells | |
| 650 | 4 | |a multiple myeloma | |
| 650 | 4 | |a stability and function of CAR-T cells | |
| 650 | 4 | |a timepoint | |
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