靶向CD7嵌合抗原受体T细胞的功能验证及改进的初步探索  

Functional validation and improvement of chimeric antigen receptor T cells targeting CD7

作　　者：张祎 王佳茜 张蕊[2] 赵明峰[1,2] Zhang Yi;Wang Jiaxi;Zhang Rui;Zhao Mingfeng(Department of Hematology,First Center Clinical School of Medicine,Tianjin Medical University,Tianjin 300380,China;Department of Hematology,Tianjin First Central Hospital,Tianjin 300380,China)

机构地区：[1]天津医科大学一中心临床医学院血液科,天津300380 [2]天津市第一中心医院血液科,天津300380

出　　处：《中华微生物学和免疫学杂志》2024年第11期926-934,共9页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金面上项目(81970180);天津市多元投入基金重点项目(21JCZDJC01240);天津市科技支撑重点项目(20YFZCSY00800);天津市医学重点学科(专科)建设项目(TJYXZDXK-056B)。

摘　　要：目的验证蛋白质阻滞技术处理的靶向CD7的嵌合抗原受体T细胞(chimeric antigen receptor T cells,CAR-T)的有效性,并使用达沙替尼预处理CD7 CAR-T细胞,验证可否增强CD7 CAR-T细胞杀伤能力或者逆转耗竭表型。方法将绿色荧光蛋白(green fluorescent protein,GFP)标记的肿瘤细胞与CD7 CAR-T细胞或T细胞按不同的效靶比以相同的培养体积和相同CAR-T/T细胞数量共孵育。流式细胞仪检测肿瘤细胞数量,并计算CAR-T细胞对肿瘤细胞的杀伤作用。通过尾静脉注射1×106个表达荧光素酶的CCRF-CEM细胞构建T系急性淋巴细胞白血病小鼠模型,评价CD7 CAR-T细胞的治疗效果。结果CD7 CAR-T细胞对CCRF-CEM、Jurkat细胞有明显杀伤效果,而对CD7阴性的NALM6细胞没有杀伤作用。与未转导的T细胞治疗组小鼠相比,CD7 CAR-T细胞治疗组小鼠的体内肿瘤负荷明显降低,存活时间显著延长(P<0.05)。达沙替尼预处理可以明显逆转CD7 CAR-T的耗竭表型(P<0.05),并且不会对杀伤效果和增殖能力产生不良影响。结论蛋白质阻滞技术处理的CD7 CAR-T细胞可以免于自相残杀,并且达沙替尼预处理有望提高CD7 CAR-T细胞的疗效及持久性。Objective To validate the efficacy of chimeric antigen receptor T cells targeting CD7(CD7 CAR-T cells)modified with protein blocking technology and analyze whether pretreatment with dasatinib can enhance CD7 CAR-T killing ability or reverse the depletion phenotype.Methods Green fluorescent protein(GFP)-labeled tumor cells were co-incubated with CD7 CAR-T cells or T cells at different potency-to-target ratios,but the culture volume and the numbers of CAR-T/T cells were same.The number of tumor cells was detected using flow cytometry.The killing effect of CAR-T cells on tumor cells was evaluated.A mouse model of acute T-lymphoblastic leukemia(T-ALL)was constructed by injecting 1×106 luciferase-expressing CCRF-CEM cells into the mouse tail vein to evaluate the therapeutic effect of CD7 CAR-T cells.Results CD7 CAR-T cells had a significant killing effect on CCRF-CEM and Jurkat cells,but not on CD7-negative NALM6 cells.The mice in the group receiving CD7 CAR-T cells had a significantly reduced in vivo tumor load and a significantly prolonged survival time as compared with the mice in the group receiving untransduced T cells(P<0.05).Dasatinib pretreatment significantly reversed the depletion phenotype of CD7 CAR-T cells(P<0.05)and had no adverse effects on the killing effect and the proliferation of the cells.Conclusions Protein-blocking technology-modified CD7 CAR-T cells are protected from killing each other,and pretreatment with dasatinib is expected to improve the efficacy and durability of CD7 CAR-T cells.

关 键 词：嵌合抗原受体T细胞 CD7 达沙替尼 功能验证 

分 类 号：R392[医药卫生—免疫学]