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| 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 |
| 马宜冬1△,王真2△,巩睿智2,李林芳3,吴红平3,金华君3*,钱其军3* |
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(1. 苏州大学医学部基础医学与生物科学学院细胞生物学系, 苏州 215123;
2. 第二军医大学东方肝胆外科医院生物治疗科, 上海 200438;
3. 第二军医大学东方肝胆外科医院基因-病毒治疗实验室, 上海 200438
△共同第一作者
*通信作者) |
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| 摘要: |
| 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。 |
| 关键词: 嵌合抗原受体 黏蛋白1 Jurkat细胞 肝细胞癌 |
| DOI：10.3724/SP.J.1008.2014.01177 |
| 投稿时间：2014-04-12修订日期：2014-08-06 |
| 基金项目:艾滋病和病毒肝炎等重大传染病的防治重大专项(2013ZX10002-010-007);上海工程技术研究中心专项课题(12DZ2251600). |
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| Specific cytotoxicity of MUC1 chimeric antigen receptor-engineered Jurkat T cells against hepatocellular carcinoma |
| MA Yi-dong1△,WANG Zhen2△,GONG Rui-zhi2,LI Lin-fang3,WU Hong-ping3,JIN Hua-jun3*,QIAN Qi-jun3* |
(1. Department of Cell Biology, School of Biology and Basic Medical Sciences, Medical College, Soochow University, Suzhou 215123, Jiangsu, China;
2. Tumor Immunology and Gene Therapy Center, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai 200438, China;
3. Laboratory of Viral and Gene Therapy, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai 200438, China
△Co-first authors.
*Corresponding authors) |
| Abstract: |
| Objective To investigate the specific cytotoxicity of MUC1-specific chimeric antigen receptor (CAR)-engineered Jurkat T cells against 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 types of MUC1-specific CARs-engineered Jurkat T cells could recognize MUC1 molecule and specifically kill MUC1 over-expressed HCC cells while left 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(P<0.05 or 0.01). Conclusion The MUC1-sepcific CAR-engineered Jurkat T cells can specifically kill MUC1 over-expressed HCC cells. |
| Key words: chimeric antigen receptors mucin 1 Jurkat cells hepatocellular carcinoma |
