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[CD7嵌合抗原受体修饰的NK-92MI细胞系对CD7阳性血液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].

作者信息

Zhu Xin-Ying, Liu Xin, Wang Xing-Bing, Wang An-You, Wang Min, Liu Na-Na, You Feng-Tao, Pan Gui-Fang, Yang Lin

机构信息

Department of Hematology, Anhui Provincial Hospital, Provincial Hospital Affiliated to Anhui Medical University, Hefei 230001, Anhui Province, China.

Department of Hematology, Anhui Provincial Hospital, Provincial Hospital Affiliated to Anhui Medical University, Hefei 230001, Anhui Province, China,E-mail:

出版信息

Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2020 Aug;28(4):1367-1375. doi: 10.19746/j.cnki.issn.1009-2137.2020.04.049.

Abstract

OBJECTIVE

To investigate the killing effect of NK-92MI 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-92MI and CD7-CAR-NK-92MI cells, killing efficiencies of CD7-CAR-NK-92MI cells to CD7 hematological tumor cells in vitro were determined by flow cytometry using PE Annexin V Apoptosis Detection Kit. Secretion differences of NK-92MI and CD7-CAR-NK-92MI 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-92MI cells to CD7 T-ALL, AML, T-cell lymphoma tumor cells were significantly higher than those of NK-92MI cells without genetical modification. The difference showed statistically significant (P<0.05). The level of IFN-γ and granzyme B were significantly increased among cytokines secreted by CD7-CAR-modified NK-92MI cells as compared with those of NK-92MI cells without genetical modification (P<0.05) .

CONCLUSION

CD7-CAR-modified NK-92MI 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.

摘要

目的

研究经嵌合抗原受体(CD7-CAR)修饰且特异性靶向CD7的NK-92MI细胞对CD7血液系统恶性肿瘤细胞的杀伤作用。

方法

收集5例CD7急性T淋巴细胞白血病(T-ALL)、10例急性髓系白血病(AML)和6例T细胞淋巴瘤这三种类型的血液系统恶性肿瘤细胞,经离心、培养后用于检测肿瘤细胞表面靶点的表达水平;采用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凋亡检测试剂盒通过流式细胞术测定CD7-CAR-NK-92MI细胞对CD7血液系统肿瘤细胞的体外杀伤效率。使用CBA试剂盒评估NK-92MI和CD7-CAR-NK-92MI细胞因子白细胞介素(IL)-2、干扰素(IFN)-γ和颗粒酶B检测的分泌差异。

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显著高于未基因修饰的NK-92MI细胞。差异具有统计学意义(P<0.05)。与未基因修饰的NK-92MI细胞相比,CD7-CAR修饰的NK-92MI细胞分泌的细胞因子中IFN-γ和颗粒酶B水平显著升高(P<0.05)。

结论

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,并显示出特异性靶向作用,为CD

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