中国肿瘤生物治疗杂志2025，Vol.32Issue(4)：356-363,8.DOI:10.3872/j.issn.1007-385x.2025.04.002
局部分泌IL-2的CD19 CAR-T细胞构建与功能
Construction and functional study of CD19 CAR-T cells that secrete IL-2 locally
摘要
Abstract
Objective:To investigate the impact of locally IL-2-secreting CAR-T cells(Syn.CD19.IL-2 CAR-T cells)constructed using a novel single-vector SynNotch system on the function of conventional CD19 CAR-T cells.Methods:Building upon our previously established single-vector SynNotch system,the CD19-specific FMC63 antibody was integrated with an IL-2 expression module to prepare Syn.CD19.IL-2 CAR-T cells through T cell transduction using a self-inactivating retroviral vector.For validation experiments,cells were divided into the Syn.CD19.IL-2 CAR-T cell group and the untransduced T cells group.ELISA was used to detect IL-2 secretion levels after antigen stimulation.The CFSE staining assay was implemented to evaluate the effects of IL-2 secretion on the proliferation of CD19 CAR-T cells when CD19 CAR-T cells were co-cultured with Raji-Luc or SW620-CD19-Luc cells in the presence of Syn.CD19.IL-2 CAR-T cells.Flow cytometry was employed to detect CD69 expression and monitor the activation status of CD19 CAR-T cells when co-cultured with Raji-Luc cells in the condition of Syn.CD19.IL-2 CAR-T cells secreting IL-2.Results:The self-inactivating retroviral vector Syn.CD19.IL-2 CAR were successfully constructed and Syn.CD19.IL-2 CAR-T cells with a viral titer>1×10⁷ copies/mL and transduction efficiency of 37.1%were generated.After antigen stimulation,SynNotch CAR-T cells demonstrated significantly elevated IL-2 secretion compared with the untransduced T cell group(P<0.001).When Syn.CD19.IL-2 CAR-T cells secreted IL-2,the CD19 CAR-T cells exhibited enhanced proliferative capacity and elevated activation level(all P<0.001).Conclusion:Successfully constructed Syn.CD19.IL-2 CAR-T cells can significantly enhance the proliferative capacity and activation ability of conventional CD19 CAR-T cells.关键词
SynNotch系统/CAR-T细胞/IL-2/CD19Key words
SynNotch system/CAR-T cell/IL-2/CD19分类
临床医学引用本文复制引用
王文娟,陈静,胡周,刘秀盈,王建勋..局部分泌IL-2的CD19 CAR-T细胞构建与功能[J].中国肿瘤生物治疗杂志,2025,32(4):356-363,8.基金项目
王建勋高层次人才科研启动经费(No.90011451310032) （No.90011451310032）
