中国肿瘤生物治疗杂志2025，Vol.32Issue(11)：1181-1187,7.DOI:10.3872/j.issn.1007-385x.2025.11.011
靶向CD7抗原的嵌合共刺激受体修饰的γδ T细胞的制备方法及初步功能鉴定
Preparation method and preliminary functional characterization of chimeric costimulatory receptor-modified γδ T cells targeting the CD7 antigen
摘要
Abstract
Objective:To develop a chimeric costimulatory receptor(CCR)targeting the CD7 antigen and prepare CCR-modified γδ T cells from healthy donors for the evaluation of its in vitro and in vivo cytotoxic effects against T-cell acute lymphoblastic leukemia(T-ALL)cells.Methods:Lentiviral vectors carrying CD7-DAP10-CCR were constructed and γδ T cells in the peripheral blood of healthy individuals were transduced to prepare CCR γδ T cells targeting the CD7 antigen(CD7-DAP10-CCR-γδ T).The obtained cells were expanded in vitro using artificial antigen-presenting cells(aAPC)expressing CD64,CD86,and CD137L.The in vitro cytotoxic effects of CD7-DAP10-CCR-γδ T cells against T-ALL cells(Jurkat),CD7-deficient Jurkat cells(CD7⁻ Jurkat),and healthy donor primary αβ T cells were detected using the Annexin Ⅴ/7-AAD assay.The experiment was performed at three effector-to-target(E∶T)ratios(1∶1,1∶3,and 1∶10),with Jurkat cells as CD7 positive target cells,CD7⁻ Jurkat cells as CD7 negative target cells to verify the killing specificity,and healthy donor primary αβ T cells as CD7 positive normal control cells to evaluate the off-target effects of CD7-DAP10-CCR-γδ T cells.The incubation time was 18-24 h.Furthermore,the in vivo efficacy was evaluated in an immunodeficient mouse model bearing T-ALL xenografts.In vivo imaging of tumor-bearing immunodeficient mice was regularly conducted,their body weight and length of survival monitored to evaluate in vivo efficacy of CD7-DAP10-CCR-γδ T cells in tumor-bearing immunodeficient mice.Results:CD7-DAP10-CCR-γδ T cells were successfully prepared in vitro using aAPC,achieving an average expansion fold exceeding 10 000.In vitro cytotoxicity assays demonstrated that these cells exhibited significantly high killing activity against T-ALL cells and significantly high toxicity against Jurkat cells(P<0.01),while showing limited cytotoxicity against CD7⁻ Jurkat cells and negligible effects on normal primary CD7-high αβ T cells.In vivo efficacy experiment on tumor-bearing immunodeficient mice indicated that treatment with CD7-DAP10-CCR-γδ T cells resulted in a significant prolongation of survival compared with the PBS control group.Conclusion:CD7-DAP10-CCR-γδ T cells can be successfully generated in vitro using aAPC.CD7-DAP10-CCR-γδ T cells demonstrate strong cytotoxicity against T-ALL cells both in vitro and in vivo,which suggests therapeutic potential of CD7-DAP10-CCR-γδ T cells against T-ALL cells.关键词
急性T淋巴细胞白血病/γδ T细胞/人工抗原提呈细胞/CD7-DAP10-CCR-γδ T细胞/嵌合共刺激受体/Jurkat细胞/K562细胞Key words
T-lineage acute lymphoblastic leukemia(T-ALL)/γδ T cell/artificial antigen presenting cell(aAPC)/CD7-DAP10-CCR-γδ T cell/chimeric costimulatory receptor(CCR)/Jurkat cell/K562 cell分类
医药卫生引用本文复制引用
汪敏,张萍,游凤涛,许汉英,杨林..靶向CD7抗原的嵌合共刺激受体修饰的γδ T细胞的制备方法及初步功能鉴定[J].中国肿瘤生物治疗杂志,2025,32(11):1181-1187,7.基金项目
国家生物药技术创新中心"揭榜挂帅"技术攻关项目(No.NCTIB2023XB01003) （No.NCTIB2023XB01003）
安徽省科技重大专项项目(No.202203a07020030) （No.202203a07020030）
安徽省重点研究与开发计划项目(No.2023s07020010) （No.2023s07020010）
苏州市科技计划项目(No.SYG2024066) （No.SYG2024066）
