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In vitro expansion of NK cells by combination of CD137 monoclonal antibody and IL-15 and their cytotoxicity against lung cancer cells
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Abstract:
Objective:To explore an in vitro expansion method of NK cells by CD137 monoclonal antibody combination with IL-15, and to study the cytotoxicity of expanded NK cells against lung cancer cells. Methods:CD3-CD56+NK cells were isolated from peripheral blood mononuclear cells (PBMCs) using negative MACS, and divided into five groups: a control group, IL-2, IL-2+ CD137mAb，IL-2+IL-15, and IL-2+IL-15+ CD137mAb groups. The expansion, phenotype and cytotoxicity of lung cancer A549 cells and IFN-γ secretion of NK cells were evaluated by MTT, FACS, LDH, and ELISA, respectively. Results: The purity of NK cells increased to (93.28+3.21)% after MACS sorting. In group IL-2+IL-15+CD137mAb, the NK cells were expanded obviously higher than were those in IL-2+CD137mAb，IL-2+IL-15，IL-2 and control groups（86.20±5.00 vs 60.01±5.00, 49.06±4.39, 17.04±1.49, 3.95+0.23, P<0.01). The cytotoxicity of expanded NK cells in group IL-2+IL-15+CD137mAb was significantly higher than that in the other groups (\[93.14±327\]% vs \[83.15±403\]%, \[7125±3.24\]%, \[62.27±3.01\]%, \[49.38±2.35\]%, P<0.01). In group IL-2+IL-15+CD137mAb, the IFN-γ level in cell supernatant was significantly higher than that in IL-2+CD137mAb, IL-2+IL-15, IL-2 and control groups (\[296.25±9.79\] vs \[260.47±11.55\], \[201.13±6.36\], \[138.36±6.09\], \[38.42±3.56\] pg/ml; P<0.01).Conclusion: CD137 monoclonal antibody combination with IL-15 can efficiently expand NK cells with an effective cytotoxicity on lung cancer A549 cells.
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Project Supported:
Project supported by the Science and Technology Foundation of Suzhou (No. 510307)
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