靶向黏蛋白1嵌合抗原受体修饰的Jurkat T细胞特异杀伤肝癌细胞

目的探讨靶向黏蛋白1(mucin 1,MUC1)的嵌合抗原受体(chimeric antigen receptor,CAR)修饰的T细胞株(CAR-T)对MUC1高表达肝癌细胞的特异杀伤能力。方法分别构建第1代与第3代靶向MUC1的CAR基因表达框(MUC1-CAR与G3MUC1-CAR),并将其装入慢病毒载体,利用获得的重组慢病毒感染Jurkat细胞,通过CCK-8法、ELISA法、乳酸脱氢酶释放实验,分别检测在MUC1抗原存在条件下,CAR-T细胞的增殖、IL-2分泌量、杀伤作用。结果成功构建2种靶向MUC1嵌合抗原受体表达框的重组慢病毒Jurkat CAR-T细胞株。两种Jurkat CAR-T细胞均能特异识别MUC1分子,杀伤MUC1高表达的肝癌细胞QGY-7701,而对MUC1低表达的正常肝细胞基本不杀伤;第3代CAR修饰的Jurkat T细胞接触MUC1分子后,其增殖速度、IL-2分泌量和杀伤效率均优于第1代MUC1修饰细胞(P<0.05或P<0.01)。结论靶向MUC1的Jurkat CAR-T细胞能特异杀伤MUC1高表达的肝癌细胞。

Specific cytotoxicity of MUC1 chimeric antigen receptor-engineered Jurkat T cells against hepatocellular carcinoma

Objective To investigate the killing effect of MUC1-specific chimeric antigen receptor (CAR)-engineered Jurkat T cells on hepatocellular carcinoma (HCC) cells. Methods The expression cassettes of both the first and the third generation MUC1-specific CAR gene (i.e. MUC1-CAR and G3MUC1-CAR) were constructed and cloned into lentivirus transfer plasmids, respectively. Then, the obtained recombinant lentiviruses carrying the MUC1-specific CAR gene and hrGFP reporter genes were used to infect Jurkat T cells in vitro to establish MUC1-sepcific CAR-engineered Jurkat cells (i.e. CAR-T cells). Subsequently, the assays of CCK-8, ELISA, and LDH were used to detect the cell proliferation, IL-2 secretion and killing effect of the CAR-T cells, respectively. Results We successfully constructed the expression cassettes of MUC1-sepcific CAR gene and the corresponding recombinant lentivirus, established the MUC1-specific CAR-engineered Jurkat T cells. Both MUC1-specific CARs-engineered Jurkat T cells could recognize MUC1 molecule and specifically kill MUC1 over-expression HCC cells while leave normal hepatic cells almost undamaged. In addition, the cell proliferation, the secretion of IL-2 and killing effect of the G3MUC1-CAR-engineered Jurkat T cells was significantly superior to the MUC1-CAR-engineered counterpart. Conclusion The MUC1-sepcific CAR-engineered Jurkat T cells can specifically kill MUC1 over-expression HCC cell.