Comparison of FACS and PCR for detection of BCMA-CAR-T cells
Chimeric-antigen-receptor (CAR)-T-cell therapy is already widely used to treat patients who are relapsed or refractory to chemotherapy, antibodies, or stem-cell transplantation. Multiple myeloma still constitutes an incurable disease. CAR-T-cell therapy that targets BCMA (B-cell maturation antigen)...
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| Main Authors: | , , , , , , , , , , , , , , , |
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| Format: | Article (Journal) |
| Language: | English |
| Published: |
14 January 2022
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| In: |
International journal of molecular sciences
Year: 2022, Volume: 23, Issue: 2, Pages: 1-14 |
| ISSN: | 1422-0067 |
| DOI: | 10.3390/ijms23020903 |
| Online Access: | Verlag, kostenfrei, Volltext: https://doi.org/10.3390/ijms23020903 Verlag, kostenfrei, Volltext: https://www.mdpi.com/1422-0067/23/2/903 
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| Author Notes: | Avinoam Reichman, Alexander Kunz, Jara J. Joedicke, Uta E. Höpken, Anna Keib, Brigitte Neuber, David Sedloev, Lei Wang, Genqiao Jiang, Angela Hückelhoven-Krauss, Franziska Eberhardt, Carsten Müller-Tidow, Martin Wermke, Armin Rehm, Michael Schmitt and Anita Schmitt |
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| 520 | |a Chimeric-antigen-receptor (CAR)-T-cell therapy is already widely used to treat patients who are relapsed or refractory to chemotherapy, antibodies, or stem-cell transplantation. Multiple myeloma still constitutes an incurable disease. CAR-T-cell therapy that targets BCMA (B-cell maturation antigen) is currently revolutionizing the treatment of those patients. To monitor and improve treatment outcomes, methods to detect CAR-T cells in human peripheral blood are highly desirable. In this study, three different detection reagents for staining BCMA-CAR-T cells by flow cytometry were compared. Moreover, a quantitative polymerase chain reaction (qPCR) to detect BCMA-CAR-T cells was established. By applying a cell-titration experiment of BCMA-CAR-T cells, both methods were compared head-to-head. In flow-cytometric analysis, the detection reagents used in this study could all detect BCMA-CAR-T cells at a similar level. The results of false-positive background staining differed as follows (standard deviation): the BCMA-detection reagent used on the control revealed a background staining of 0.04% (±0.02%), for the PE-labeled human BCMA peptide it was 0.25% (±0.06%) and for the polyclonal anti-human IgG antibody it was 7.2% (±9.2%). The ability to detect BCMA-CAR-T cells down to a concentration of 0.4% was similar for qPCR and flow cytometry. The qPCR could detect even lower concentrations (0.02-0.01%). In summary, BCMA-CAR-T-cell monitoring can be reliably performed by both flow cytometry and qPCR. In flow cytometry, reagents with low background staining should be preferred. | ||
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