Efficient amplification of NK cells and their anti-tumor cytotoxic activity in vitro
10.3969/j.issn.1000-484X.2017.08.013
- VernacularTitle:NK细胞的高效扩增及其对体外肿瘤模型的杀伤效果研究
- Author:
Chunxiang LIU
;
Yi ZHANG
;
Huiying LU
- Keywords:
NK cells;
Tumor cells;
Cytotoxic activity;
Amplification
- From:
Chinese Journal of Immunology
2017;33(8):1186-1190,1196
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To efficiently amplify NK cells and determine their cytotoxic activity against a variety of tumor cell lines in vitro,thereby providing evidence for potential clinical application.Methods: PBMCs were isolated from adult peripheral blood and co-cultured with K562 cells that were genetically modified to express 4-1BBL,IL-15 and IL-21 on the surface for 15 days to effectively amplify NK cells.The total cell number and Purity of CD3-CD56+ cells were measured.Granzyme B and perforin expression of the amplified NK cells were detected by flow cytometry and real-time PCR.The anti-tumor effect on different cancer cells was evaluated.Results: This method obtained a more than 1.1×1010 CD3-CD56+ NK cells with 95% purity over a 15 day amplification procedure.The expanded NK cells could efficiently release granzyme B and Perforin.The cytotoxicity against different tumor cells was followed the order from strong to weak:gastric,pancreatic,cervical,ovarian and renal cancer cells,with the highest activity against gastric cancer cell line A549 (90% at E∶T=10∶1) (P<0.05).A time-dependent killing effect of activated NK cells on cervical,liver and pancreatic cancer cells was observed.Conclusion: This amplification procedure can consistently generate large amounts of pure NK cells with effective cytotoxic function against a variety of tumor cells.
