细胞毒性
嵌合抗原受体
NKG2D公司
细胞生物学
HEK 293细胞
嘌呤霉素
白细胞介素12
化学
报告基因
K562细胞
绿色荧光蛋白
分子生物学
癌症研究
细胞培养
生物
细胞毒性T细胞
基因表达
免疫学
免疫系统
T细胞
体外
基因
遗传学
蛋白质生物合成
作者
Changjiang Guo,Han Chen,Jie Yu,Hui Lü,Qing Xia,Xiaojuan Li,Xiali Guo,Tong Wang,Lin Zhi,Zhiyuan Niu,Wuling Zhu
标识
DOI:10.1016/j.molimm.2023.01.010
摘要
Adoptive chimeric antigen receptor (CAR)-modified T or NK cells (CAR-T/NK) have emerged as a novel form of disease treatment. Lentiviral vectors (LVs) are commonly employed to engineer NK cells for the efficient expression of CARs. This study reported the influence of single-promoter and dual-promoter LVs on the CAR expression and cytotoxicity of engineered NK cells. We constructed a third-generation NKG2D-based CAR that kills cancer cells by targeting up to eight stress-induced ligands (NKG2DLs). Our results demonstrated that the CAR exhibits both a higher expression level and a higher coexpression concordance with the GFP reporter in HEK-293T or NK92 cells by utilizing the optimized single-promoter pCDHsp rather than the original dual-promoter pCDHdp. After puromycin selection, the pCDHsp produces robust CAR expression and enhanced in vitro cytotoxicity of engineered NK cells. Therefore, infection with a single-promoter pCDHsp lentivector is recommended to prepare CAR-engineered NK cells. This research helps to optimize the production of CAR-NK cells and enhance their functional activity, to provide CAR-NK cell products with better and more uniform quality.
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