TY - CHAP
T1 - Basic procedures for detection and cytotoxicity of chimeric antigen receptors
AU - Mihara, Keichiro
AU - Yoshida, Tetsumi
AU - Bhattacharyya, Joyeeta
PY - 2019/1/1
Y1 - 2019/1/1
N2 - Chimeric antigen receptors against CD19 (anti-CD19-CAR) are widely recognized and used by not only researchers associated with immunology, molecular biology, and cell biology but also physicians to treat B-cell malignancies. Anti-CD19-CAR is currently clinically available as one of the therapeutic modalities for refractory acute B-cell-typed lymphoblastic leukemia (B-ALL) patients. However, to detect CAR on the cell surface and investigate the efficacy of CAR-T cells, there are numerous experimental modalities including flow cytometry, the Cr-releasing assay, immunoblot, and immunostaining. We have chosen several techniques, which are necessary and sufficient as well as reliable and reproducible to detect and assess the killing effect of CAR-T cells. Here, we describe protocols for basic experiments and procedures for the detection of CAR on transduced cells and in in vitro coculture experiments to assess cytotoxicity using CAR-T cells.
AB - Chimeric antigen receptors against CD19 (anti-CD19-CAR) are widely recognized and used by not only researchers associated with immunology, molecular biology, and cell biology but also physicians to treat B-cell malignancies. Anti-CD19-CAR is currently clinically available as one of the therapeutic modalities for refractory acute B-cell-typed lymphoblastic leukemia (B-ALL) patients. However, to detect CAR on the cell surface and investigate the efficacy of CAR-T cells, there are numerous experimental modalities including flow cytometry, the Cr-releasing assay, immunoblot, and immunostaining. We have chosen several techniques, which are necessary and sufficient as well as reliable and reproducible to detect and assess the killing effect of CAR-T cells. Here, we describe protocols for basic experiments and procedures for the detection of CAR on transduced cells and in in vitro coculture experiments to assess cytotoxicity using CAR-T cells.
UR - http://www.scopus.com/inward/record.url?scp=85058593851&partnerID=8YFLogxK
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U2 - 10.1007/978-1-4939-8958-4_13
DO - 10.1007/978-1-4939-8958-4_13
M3 - Chapter
C2 - 30539476
AN - SCOPUS:85058593851
T3 - Methods in Molecular Biology
SP - 299
EP - 306
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -