Helmholtz Gemeinschaft


Comparison of FACS and PCR for detection of BCMA-CAR-T cells

PDF (Original Article) - Requires a PDF viewer such as GSview, Xpdf or Adobe Acrobat Reader

Item Type:Article
Title:Comparison of FACS and PCR for detection of BCMA-CAR-T cells
Creators Name:Reichman, A. and Kunz, A. and Joedicke, J.J. and Höpken, U.E. and Keib, A. and Neuber, B. and Sedloev, D. and Wang, L. and Jiang, G. and Hückelhoven-Krauss, A. and Eberhardt, F. and Müller-Tidow, C. and Wermke, M. and Rehm, A. and 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
Page Range:903
Date:14 January 2022
Official Publication:https://doi.org/10.3390/ijms23020903
PubMed:View item in PubMed

Repository Staff Only: item control page


Downloads per month over past year

Open Access
MDC Library