Construction of anti-BCMA chimeric antigen receptor (CAR-BCMA) modified T cells and its cytotoxicity against tumor cells
10.3872/j.issn.1007-385x.2019.02.003
- VernacularTitle:靶向B细胞成熟抗原的嵌合抗原受体T细胞的构建及其对肿瘤细胞的杀伤
- Author:
HAO Ruidong
1
;
TIAN Fang
;
YANG Zhenli
;
WANG Minliang
1
;
ZHANG Dating
1
;
LI Yantao
1
;
FAN Pengcheng
1
;
ZHU Xuejun
;
LIU Gentao
1
Author Information
1. Shanghai Biomed-Union Biotechnology Co. Ltd. Shanghai
- Publication Type:Journal Article
- Keywords:
B cell maturation antigen (BCMA);
chimeric antigen receptor;
multiple myeloma;
IFN-γ;
cytotoxity
- From:
Chinese Journal of Cancer Biotherapy
2019;26(2):152-158
- CountryChina
- Language:Chinese
-
Abstract:
Objective： ：To explore a novel chimeric antigen receptor (CAR)-T cell treatment to treat Multiple Myeloma (MM) via target B cell maturation antigen (BCMA). Methods： ：A CAR-BCMA molecular was constructed based on mouse originated BCMA scFv, and was packaged into lentiviral vector and transfected into T cells from healthy donors to construct CAR-BCMA-T cells. The BCMApositive cell lines A549-BCMA, A549-BCMAOFP and K562-BCMA were constructed as target cells. Then, the CAR-BCMA-T cells were co-incubated with the constructed target cells and human myeloma U266 cells, and the cytotoxic effects of CAR-BCMA-T cells were evaluated via CCK-8 and FACS. Finally, the CAR-BCMA-T cells originated from MM patients were constructed, and its cytotoxicity against A549-BCMA were examined; in addition, the IFN-γ release level in CAR-BCMA-T cells was evaluated by ELISA and FACS. Results: After 11 days’incubation, the CAR-BCMA-T cells originated from healthy donors amplified 300 times with a positive rate of 43%. The BCMApositive target cell lines were constructed successfully. Under an effector : target ratio of 5:1, the killing rates of CARBCMA-T cells against A549-BCMA, K562-BCMA and U266 were about 80%, 60%, and 80%, respectively, which were significantly higher than those against BCMA negative cells; and the cytotoxicity was related to the BCMA expression level in target cells. What’ s more, at the effector : target ratio of 20:1, the CAR-BCMA-T cells originated from MM patients were demonstrated to exhibit a killing rate of more than 95% againstA549-BCMApositive cells, and produced large amount of IFN-γ. Conclusion: CAR-BCMA-T cells originated from both healthy and MM donors were successfully constructed, and they can effectively and specifically kill BCMA positive tumor cells.
- Full text:20190203.pdf
