靶向CD7的嵌合抗原受体NK-92MI细胞对CD7阳性血液系统恶性肿瘤细胞的杀伤作用  

Killing Effect of A CD7 Chimeric Antigen Receptor-Modified NK-92MI Cell Line on CD7-Positive Hematological Malignant Cells

作　　者：朱欣颖 刘欣 王兴兵 汪安友 汪敏 刘纳纳 游凤涛 潘桂芳 杨林 ZHU Xin-Ying;LIU Xin;WANG Xing-Bing;WANG An-You;WANG Min;LIU Na-Na;YOU Feng-Tao;PAN Gui-Fang;YANG Lin(Department of Hematology,Anhui Provincial Hospital,Provincial Hospital Affiliated to Anhui Medical University,Hefei 230001,Anhui Province,China;PersonGen-Anke Cell Technology Co.,Ltd,Hefei 230088,Anhui Province,China)

机构地区：[1]安徽医科大学附属省立医院,安徽省立医院血液科,安徽合肥230001 [2]博生吉安科细胞技术有限公司,安徽合肥230088

出　　处：《中国实验血液学杂志》2020年第4期1367-1375,共9页Journal of Experimental Hematology

基　　金：安徽省重点研究与开发计划项目(201904a07020102)。

摘　　要：目的:探讨特异性靶向CD7分子的嵌合抗原受体(CD7-CAR)修饰的NK-92MI细胞对CD7阳性血液系统恶性肿瘤细胞的杀伤作用。方法:分别收集5例CD7^+的急性T淋巴细胞白血病(acute T-lymphoblastic leukemia,T-ALL)、10例急性髓细胞白血病(acute myeloid leukemia,AML)和6例T细胞淋巴瘤三类血液系统恶性肿瘤细胞,经实验室离心处理、培养后检测肿瘤细胞表面靶标的表达水平;应用7-AAD、CD56-APC、CD3-FITC、IgG Fc-PE流式细胞术检测NK-92MI和CD7-CAR-NK-92MI细胞的转染效率;使用PE Annexin V Apoptosis Detection Kit检测CD7-CAR-NK-92MI细胞对CD7阳性血液肿瘤细胞的体外杀伤效率。用CBA试剂盒检测杀伤后NK-92MI和CD7-CAR-NK-92MI细胞因子白介素-2(IL-2)、干扰素(IFN-γ)及颗粒酶B的分泌差异。结果:CD7-CAR修饰的NK-92MI细胞对CD7^+T-ALL、AML、T细胞淋巴瘤肿瘤细胞的杀伤效率明显高于未被修饰的阴性对照组NK-92MI细胞,其差异具有统计学意义(P<0.05);与未被修饰的NK-92MI细胞相比,经CD7-CAR修饰的NK-92MI细胞分泌的细胞因子中,IFN-γ和颗粒酶B的水平明显升高(P<0.05)。结论:经CD7-CAR修饰的NK-92MI细胞对CD7^+T-ALL、AML和T淋巴瘤细胞的杀伤效率显著提高,且具有特异性靶向作用。Objective:To investigate the killing effect of NK-92 MI cells modified by chimeric antigen receptor(CD7-CAR)and specifically targeting CD7 to CD7^+hematological malignant cells.Methods:Three types of hematological malignant tumor cells,including 5 cases of CD7^+acute T-lymphoblastic leukemia(T-ALL),10 cases of acute myeloid leukemia(AML)and 6 cases of T-cell lymphoma were collected,centrifuged,cultured and used to detect the expression levels of tumor cell surface targets;7-AAD,CD56-APC,CD3-FITC,IgG Fc-PE flow cytometry were used to detected the transfection efficiency of NK-92 MI and CD7-CAR-NK-92 MI cells,killing efficiencies of CD7-CAR-NK-92 MI cells to CD7^+hematological tumor cells in vitro were determined by flow cytometry using PE Annexin V Apoptosis Detection Kit.Secretion differences of NK-92 MI and CD7-CAR-NK-92 MI cytokines interleukin(IL)-2,interferon(IFN)-γ,and granzyme B detection were estimated by using CBA kit.Results:The killing efficiencies of CD7-CAR-modified NK-92 MI cells to CD7^+T-ALL,AML,T-cell lymphoma tumor cells were significantly higher than those of NK-92 MI cells without genetical modification.The difference showed statistically significant(P<0.05).The level of IFN-γand granzyme B were significantly increased among cytokines secreted by CD7-CAR-modified NK-92 MI cells as compared with those of NK-92 MI cells without genetical modification(P<0.05).Conclusion:CD7-CAR-modified NK-92 MI cells have significantly improved killing efficiency against CD7^+T-ALL,AML and T lymphoma cells,and shows specific targeting effects,which provides a clinical basis for the treatment of CD7^+hematological malignancies.

关 键 词：CD7 血液系统恶性肿瘤 过继免疫治疗 嵌合抗原受体 NK-92MI 

分 类 号：R733.7[医药卫生—肿瘤] R392[医药卫生—临床医学]