ABSTRACT
Objective To study the killing effect of γδT cells on different tumor cells by inducing and expanding γδT cells in vitro.Methods Collected the peripheral venous blood of 20 healthy volunteers recruited at Xinqiao Hospital of Third Military Medical University from January to March 2018.Peripheral blood mononuclear cells (PBMCs) were induced and expanded using zoledronic acid combined with interleukin-2 (IL-2) to obtain γδT cells.The cell purity was detected on day 0,7,10,14 and 16 using flow cytometry.The killing activity was detected by CCK-8 kit.Cells cultured on the 14th day were used as effector cells,and breast cancer cell BCap-37 and liver cancer cell Bel-7402 were used as target cells,and the cell killing activity was detected at the effective target ratio (E ∶ T) of 5 ∶ 1,10 ∶ 1 and 20 ∶ 1 respectively.Results The purity of γδT cells were respectively (3.35 ± 1.32) ％,(50.76 ± 5.76) ％,(80.43 ± 4.53) ％,(90.56 ± 3.34) ％,(89.54 ± 4.42) ％ on day 0,7,10,14,16,with significant difference (F =18.431,P =0.012).The efficiency of γδT cells killing BCap-37 tumor cells were (31.3 ± 2.0) ％ (E ∶ T =5 ∶ 1),(48.7 ± 1.6) ％(E ∶T=10∶1),(71.3 ±2.4)％ (E ∶T=20 ∶ 1) respectively,with significant difference (F=16.724,P =0.016),further comparison between each two groups:10 ∶ 1 vs.5 ∶ 1 (P =0.013),20 ∶ 1 vs.5 ∶ 1 (P =0.017),20∶1 vs.10 ∶1 (P =0.011).The killing rate increased with the increase of target ratio.The efficiency of γδT cells killing Bel-7402 tumor cells were (34.5 ± 2.0) ％ (E ∶ T =5 ∶ 1),(52.4 ± 1.9) ％(E ∶ T =10 ∶ 1),(74.5 ± 1.6) ％ (E ∶ T =20 ∶ 1) respectively,with significant difference (F =18.253,P=0.013),further comparison between each two groups:10 ∶ 1 vs.5 ∶ 1 (P=0.015),20 ∶ 1 vs.5 ∶ 1 (P=0.012),20 ∶ 1 vs.10 ∶ 1 (P =0.015).The killing rate increased with the increase of target ratio.Conclusion We can obtain high purity γδT cells using zoledronic acid combined with IL-2 induced PBMCs,and the amplified γδT cells have significant killing effect on BCap-37 and Bel-7402 tumor cells.
