Abstract
Objective To observe the effect of IL-2 and IL-15 culturing with (NK) natural killer cell of peripheral blood in vitro and improve the cytotoxicity of NK cell against Wilms tumor cell line SK-NEP-1. Methods Eight patients with Wilms tumor (experiment group) and 8 normal children (control group) were recruited.For each case, peripheral blood (4 ml) was collected and mononuclear cell (PBMC) separated with Ficoll reagent.CD3-CD16+ CD56+ NK cells were detected and sorted by flow cytometry.In addition, another acquisition case and control groups in peripheral blood of 4 ml cultured with IL-2(100 U/ml) or IL-2 (100 U/ml) plus IL-15 (10 ng/ml), sorting after culturing of CD3-CD16+ CD56+ NK cells.CCK8 method was used for detecting the effect of different effector cells and target cell ratio (E∶T 5∶1, 10∶1, 20∶1) (effector cells, NK cells & target cells: SK-NEP-1) cases, the killing effect of WT with peripheral blood NK cells (cultured with IL-2, IL-15) to SK-NEP-1 cells.The expressions of TNF-α and TNF-γ of CD3-CD16+ CD56+ NK cells (pre-culture, IL-2 group, IL-2 & IL-15 groups) were detected by western blot. Results The proportion of NK cell in two group, using IL-2 and IL-15 significantly increased than before [case group: (7.3±2.4)% vs (21.2±3.2)%], control group [(4.5±1.6)% vs (10.1±3.4)%] and quantity increased respectively (70.60±22.5), (30.12±33.16) times (P<0.05). After using IL-2 and IL-15, cytotoxicity of NK cell against Wilms tumor cell line (10∶1 20∶1) obviously increased [(16.3±2.1) vs (10.5±1.6)], [(23.9±3.2) vs (13.7±2.5)](P<0.05). The expressions of TNF-α and TNF-γ (post-culture) increased than (pre-culture, IL-2 group)and significantly in IL-2 plus IL-15 group (P<0.05). Conclusions The quantity and proportion of NK cell were improved among PBMCs by IL-2 plus IL-15.Also there is enhanced cytotoxicity of NK cell against Wilms tumor cell line SK-NEP-1.The reason may be due to an up-regulation of TNF. Key words: Wilms tumor; Natural killer cell; Interleukin-2; Interleukin -15; Child
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