靶向CD19和BCMA的CAR-NK92MI细胞的构建及抗肿瘤活性研究  

Construction and antitumor activity of CAR-NK92MI cells targeting CD19 and BCMA

作　　者：余卓营 李晓瑞 尚凤琴 李申 王铁山 王建勋 YU Zhuoying;LI Xiaorui;SHANG Fengqin;LI Shen;WANG Tieshan;WANG Jianxun(School of Life Sciences,Beijing University of Chinese Medicine,Beijing 102488,China)

机构地区：[1]北京中医药大学生命科学学院,北京102488

出　　处：《现代肿瘤医学》2022年第5期785-791,共7页Journal of Modern Oncology

基　　金：王建勋高层次人才科研启动经费(编号:9011451310032)。

摘　　要：目的:探讨嵌合抗原受体(chimeric antigen receptor, CAR)结构对CAR-NK细胞特异性杀伤能力的增强情况。方法:利用逆转录病毒载体分别构建出稳定表达靶向B细胞成熟抗原(B cell maturation antigen, BCMA)与分化群抗原19(cluster of differentiation 19,CD19)CAR的CAR-NK92MI细胞,随后使用流式细胞术检测经MYC-PE抗体染色的CAR-NK细胞的CAR阳性率。采用荧光素酶检测法、IncuCyte实时动态细胞成像分析系统、流式细胞术检测两种CAR-NK92MI细胞与只表达CD19或者只表达BCMA的肿瘤细胞共孵育一定时间后肿瘤细胞的凋亡情况。结果:成功构建出稳定高表达CAR的ANTI-CD19-CAR-NK92MI细胞系和ANTI-BCMA-CAR-NK92MI细胞系。体外杀伤实验结果显示,相比表达BCMA的肿瘤细胞,ANTI-CD19-CAR-NK92MI细胞对表达CD19的肿瘤细胞具有更高的杀伤活性;而相比表达CD19的肿瘤细胞,ANTI-BCMA-CAR-NK92MI细胞对表达BCMA的肿瘤细胞具有更高的杀伤活性。结论:CAR结构可以显著增强CAR-NK细胞的特异性杀伤能力。Objective:To investigate the specific cytotoxicity of CAR-NK cells by using CAR structure.Methods:BCMA and CD19 CAR-NK92 MI cells were constructed by retroviral vector respectively.Flow cytometry was uesd to detected the CAR-positive rate of CAR-NK cells stained with MYC-PE antibody.The cytotoxic activity of these CAR-NK92 MI cells against CD19-expressing or BCMA-expressing tumor cells were detected by EX-luciferase assay, Incucyte Live-Cell Analysis Systems and flow cytometry.Results:ANTI-CD19-CAR-NK92 MI cells and ANTI-BCMA-CAR-NK92 MI cells with stable and high expression of CAR were successfully constructed.The results of these cytotoxicity assays showed that ANTI-CD19-CAR-NK92 MI cells exhibited better effector function against CD19-expressing tumor cells, and ANTI-BCMA-CAR-NK92 MI cells exhibited better effector function against BCMA-expressing tumor cells.Conclusion:CAR can significantly improved the specific cytotoxicity of CAR-NK cells.

关 键 词：嵌合抗原受体NK细胞 特异性杀伤 分化群抗原19 B细胞成熟抗原 

分 类 号：R730.3[医药卫生—肿瘤]