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| Item Type: | Article |
|---|---|
| Title: | Comparison of FACS and PCR for detection of BCMA-CAR-T cells |
| Creators Name: | Reichman, A., Kunz, A., Joedicke, J.J., Höpken, U.E., Keib, A., Neuber, B., Sedloev, D., Wang, L., Jiang, G., Hückelhoven-Krauss, A., Eberhardt, F., Müller-Tidow, C., Wermke, M., Rehm, A., Schmitt, M. and Schmitt, A. |
| Abstract: | 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. |
| Keywords: | BCMA-CAR, Polymerase Chain Reaction, Detection Reagent, Flow Cytometry |
| Source: | International Journal of Molecular Sciences |
| ISSN: | 1422-0067 |
| Publisher: | MDPI |
| Volume: | 23 |
| Number: | 2 |
| Page Range: | 903 |
| Date: | 14 January 2022 |
| Official Publication: | https://doi.org/10.3390/ijms23020903 |
| PubMed: | View item in PubMed |
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