靶向NKG2D配体的嵌合抗原受体巨噬细胞具有肿瘤杀伤功能  

NKG2D chimeric antigen receptor-expressing macrophages demonstrate cytotoxicity against tumor cells

作　　者：戴昆 刘骏晨 刘茂玄 甘江彧 王琪 陈有海 万晓春[2,3] 于广 DAI Kun;LIU Junchen;LIU Maoxuan;GAN Jiangyu;WANG Qi;CHEN Youhai;WAN Xiaochun;YU Guang(Division of Immunology,Jinzhou Medical University,Jinzhou 121001,China;Institute of Biomedicine and Biotechnology,Shenzhen Institute of Advanced Technology,Chinese Academy of Sciences,Shenzhen 518055,China;University of Chinese Academy of Sciences,Beijing 100049,China)

机构地区：[1]锦州医科大学免疫学教研室,121001 [2]中国科学院深圳先进技术研究院生物医药与技术研究所,518055 [3]中国科学院大学,北京100049

出　　处：《免疫学杂志》2022年第12期1090-1099,共10页Immunological Journal

基　　金：国家重点研发计划(2019YFA0906100);深圳市科技创新委员会基础研究项目(JCYJ20170818164619194)。

摘　　要：目的 制备表达NKG2D嵌合抗原受体(CAR)的巨噬细胞(NKG2D CAR-M),研究其是否可以增强巨噬细胞对表达靶抗原的肿瘤细胞系的杀伤效应。方法 合成包含NKG2D的胞外段,CD8α铰链区和穿膜区,CD3ζ胞内信号域的NKG2D CAR基因,构建到腺病毒Ad5F35载体中,利用重组Ad5F35腺病毒感染人原代CD14^(+)单核细胞分化而来的巨噬细胞,制备NKG2D CAR-M。通过流式细胞术和荧光显微镜检测其嵌合抗原受体在巨噬细胞上的表达,通过荧光素酶法检测检测NKG2D CAR-M对高表达NKG2D配体的前列腺癌细胞系PC-3细胞杀伤效果。结果 我们成功制备了NKG2D CAR-M细胞,该细胞对高表达NKG2D配体的PC-3细胞具有非常显著的杀伤效果。结论 NKG2D CAR-M对高表达NKG2D配体的肿瘤细胞具有显著的杀伤效果。In this study, we generated human macrophages with NKG2D chimeric antigen receptor(NKG2D CAR-M) to investigate whether NKG2D CAR can enhance the killing effect of macrophages on tumor cell lines expressing the target antigen. We synthesized the NKG2D CAR gene containing the extracellular segment of NKG2D, the CD8α hinge region in transmembrane region and the CD3ζ intracellular signal domain. These segments were constructed into the adenovirus Ad5F35 vector, which was then used to infect human primary CD14^(+)monocyte-derived macrophages to express NKG2D CAR. Theexpression of chimeric antigen receptor on macrophageswas detected by flow cytometry and fluorescencemicroscopy. Data showed NKG2D-based CAR-M cellsexhibited significant cytolytic activity against NKG2Dligandsprostate cancer cell line PC-3 in an E/T ratio-dependent manner, but had no killing effect on theNKG2D ligands–cell line B16-F10 in vitro. Inconclusion, NKG2D CAR-M has significant killing effects on NKG2D ligands-expressing tumor cells, which lays the foundation for the development of NKG2D-basedCAR-M cells.

关 键 词：嵌合抗原受体 NKG2D 巨噬细胞 CAR-M 

分 类 号：R73-36[医药卫生—肿瘤]