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Archive > Volume 25 Issue 8 > 2018,25(8):767-771. DOI:10.3872/j.issn.1007-385X.2018.08.003 Prev Next
Basic Research
The construction of anti-CD19 chimeric receptor modified NK-92 cells and the killing effect of CD19 positive non-Hodgkin lymphoma cells
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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, ARH-77 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 on ARH-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.
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Project Supported:
Project supported by the Key Technology Innovation Team Project of Zhjiang Province（No. 2011R50018-06）
Clc Number:
