Killing effect of NK92 cells modified with CD33-CAR on CD33+ acute myeloid leukemia cells
DOI ： ： 10.3872/j.issn.1007-385x.2018.05.004
- VernacularTitle:靶向CD33的CAR修饰的NK92细胞对CD33+急性髓系白血病细胞的杀 伤作用
- Author:
LIU Yanzhong1
1
;
PAN Lijuan1
1
;
TANG Qulai1
1
;
SHI Jiangzhou1
1
;
ZHAO Wenjing1
1
;
HUO Lihong1
1
;
GU Chaojiang2
2
;
HU Guang2
2
;
LIU Huining
1
;
ZHANG Tongcun
1
,
3
Author Information
1. 1. College of Biotechnology, Tianjin University of Science and Technology, Tianjing 300457, China
2. College of Life Sciences and Health, Wuhan University of Science and Technology, Wuhan 430000, Hubei，China
3. 2. College of Life Sciences and Health, Wuhan University of Science and Technology, Wuhan 430000, Hubei，China
- Publication Type:Journal Article
- Keywords:
chimeric antigen receptor(CAR);
CD33;
NK92 cell;
CD33-CAR-NK92 cell；acute myeloid leukemia(AML)
- From:
Chinese Journal of Cancer Biotherapy
2018;25(5):462-468
- CountryChina
- Language:Chinese
-
Abstract:
[Abstract] Objective: To construct CD33-CAR modified NK92 cells based on CD33-scFv sequence, and to explore its killing effect on CD33+ AML (acute myeloid leukemia) cells. Methods: DNA fragment encoding CD33-CAR was synthesized by gene synthesis and molecular cloning technology and then cloned into lentiviral vector. Lentivirus were packaged and used to transfect NK92 cells. The transfection efficiency was detected by flow cytometry, and puromycin was used to screen NK92 cells stably expressing CD33-CAR (CD33-CAR-NK92). Killing effect of CD33-CAR-NK92 cells on AML cells in vitro was examined with calcein-AM release assays. IFN-γ secretions of NK92 cells and CD33-CAR-NK92 cells were measured by ELISA. Results: The pCDH-CD33-CAR lentiviral vector was successfully constructed. After lentiviral transfection, about 18.7% of NK92 cells express CD33-CAR (referred as CD33-CARNK92 cells). The percentage of CD33-CAR+ NK92 cells was about 86.3% after puromycin selection. In contrast to unmodified NK92 cells, significantly higher cytotoxic effect against CD33+ MOLM-13 cells was found in CD33-CAR-NK92 cells (P<0.01); however, there was no significant difference in cytotoxicity against CD33- JURKAT cells between NK92 cells and CD33-CAR-NK92 cells (P> 0.05). After co-culture at an effect-target ratio of 2∶1 for 6 hours, the level of IFN-γ secreted by the CD33-CAR modified NK92 cells was significantly higher than that of the unmodified ([190.97±11.52] vs [88.41±2.75]pg/ml, P<0.01). Conclusion: The CD33-CARNK92 cells could specifically recognize CD33 antigen and kill CD33+ AML cells in comparison with the unmodified NK92 cells, which provides experimental basis for clinical transformation of CD33-CAR-NK92 cells in treatingAML.
- Full text:20180504.pdf
