Construction of Second-and Third-generation CAR-T Cells Targeting BCMA and Analysis of Their Antitumor Activity in Vitro
ZHANG Weiguang1,2, WANG Keke2, WU Fangnan2, LIU Yu2, YIN Changlin1, GAO Jimin2*
This study constructs a lentiviral expression vector of the second-generation Anti-BCMA CAR containing 4-1BB or ICOS by genetic engineering, and a lentiviral expression vector of the third-generation AntiBCMA CAR containing both of these costimulatory signaling molecules. By transducing human CD3+ T cells with these lentiviral particles, the second- and third-generation Anti-BCMA CAR-T cells are obtained, respectively. Our results showed that the second-generation CAR-T cells with ICOS and 4-1BB have similar fucnctions on antineoplastic activity. Whereas, when effector/target was 2׃1, the cells with ICOS showed better killing effect to U266- Lucgfp cells comparing with 4-1BB. The third-generation Anti-BCMA CAR-T cells executed lower cytolytic capacity against U266-lucgfp cells than the second-generation CAR-T cells at the ratio of effectors/targets were 2׃1, 5׃1 and 10׃1. But when the ratio of effectors/targets was 20׃1, the specific lysis rates of the second- or thirdgeneration CAR-T cells against U266-lucgfp cells were both above 90%, which were significantly higher than those of control T cells. In conclusion, this study successfully constructed the second- and third-generation Anti-BCMA CAR-T cells, which efficiently killed highly-expressing-BCMA tumor cells. And the second-generation CAR-T cells targeting BCMA had stronger killing effect on tumor cells than the third-generation CAR-T cells. Besides, the second-generation CAR-T cells targeting BCMA had more potent killing effect on tumor cells than the third-generation CAR-T cells.
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