The construction of anti-CD19 chimeric receptor modified NK-92 cells and the killing effect of CD19 positive non-Hodgkin lymphoma cells
10.3872/j.issn.1007-385x.2018.08.003
- VernacularTitle:抗CD19嵌合受体修饰的NK-92细胞的构建及其对CD19阳性非霍奇金 淋巴瘤细胞的杀伤作用
- Author:
JIANG Xin
1
;
ZHU Shuangyue
2
;
ZHENG Haili
1
;
LIU Bingyu
3
Author Information
1. Pharmacy Department, Children's Hospital, Zhejian University School of Medicine
2. Department of Integrated Chinese and Western Medicine, Hangzhou Seventh People's Hospital
3. Department of Cardiology, Zhejiang Hospital
- Publication Type:Journal Article
- Keywords:
non-Hodgkin's lymphoma;
chimeric receptor;
NK-92;
cytotoxicity
- From:
Chinese Journal of Cancer Biotherapy
2018;25(8):767-771
- CountryChina
- Language:Chinese
-
Abstract:
Objective: A second generation CAR-NK-92 cell line expressing CD19 was constructed to investigate its specific killing effect on CD19 positive non-Hodgkin lymphoma cells. Methods: First, build CD19-CAR gene expression vector and packaged slow virus particles, then the infection rate was detected by flow cytometry after infected NK-92 cells and positive cells were further separated. Finally, detected the expression of CD19-CAR in NK-92 cells by Western blotting. U-266 with CD19 negative myeloma cells,ARH77 and HS-Sultan with CD19 positive non-Hodgkin’s lymphoma cells as target cells, and CD19CAR-NK-92 as effector cells, then the killing rate was calculated by the absolute number of tumor cells alive in the cell killing experiment. Results: Construct lentivirus vector pLVX-CD19-CAR and packaged virus particles successfully, the purity of CD19-CAR-NK-92 cells also was over 90% after infected with NK-92 cells; and Western blotting analysis showed that CD19-CAR had been successfully expressed in NK-92 cell. The killing effect of CD19CAR-NK-92 onARH-77 ([70.10±1.86]% vs [1.95±0.63]%, P<0.01) and HS-Sultan ([74.98±1.60]% vs [0.58±1.49]%, P< 0.01) cells was significantly higher than the empty vector control group of ZsGreen-NK-92, but there was no difference in killing U266 (P>0.05). Conclusion: The NK-92 cell lines expressing CD19CAR were successfully constructed, and also has specific killing effects on CD19 positive non-Hodgkin lymphoma cells.
- Full text:20180803.pdf
