IL-2、IL-15增强免疫编辑后NK细胞NKG2D的表达及其对鼻咽癌CNE2细胞的杀伤活性

IL-2 and IL-15 up-regulate NKG2D expression and enhance cytotoxicity of edited-NK cells against nasopharyngeal carcinoma cells

目的:观察IL-2、IL-15对免疫编辑后NK细胞NKG2D的表达及其对鼻咽癌CNE2细胞杀伤活性的影响。方法:抗CD56磁珠纯化NK细胞后分为4组:(1)编辑前NK细胞组:加入100 U/ml IL-2;(2)单纯编辑组:NK细胞与CNE2细胞10∶1混合,加入100U/ml IL-2;(3)IL-2再培养组:纯化编辑后的NK细胞,加入1 000 U/ml IL-2;(4)IL-15再培养组:纯化编辑后的NK细胞,加入10 ng/ml IL-15。24 h后,流式细胞仪检测各组NK细胞表面NKG2D的表达;LDH释放测定法测定效靶比20∶1时各组NK细胞对CNE2细胞的杀伤活性。结果:编辑前NK细胞组、单纯编辑组、IL-2再培养组、IL-15再培养组NK细胞表面NKG2D的表达率分别为(97.63±0.83)%、(53.50±1.25)%、(94.47±1.00)%、(98.07±0.21)%。IL-2、IL-15再培养组NK细胞NKG2D的表达分别比单纯编辑组有显著的增加(P<0.01),该4组NK细胞对CNE2细胞的杀伤活性分别为(35.90±3.27)%、(4.70±2.30)%、(31.70±3.56)%、(40.18±2.94)%,IL-2再培养组、IL-15再培养组明显提高编辑后NK细胞对CNE2细胞的杀伤活性(P<0.01),IL-15的作用强于IL-2。结论:高剂量IL-2、IL-15可以上调免疫编辑后NK细胞表面NKG2D的表达,恢复编辑后NK细胞对鼻咽癌细胞CNE2的杀伤活性,IL-15的作用强于IL-2。

Objective： To study the effects of IL-2 and IL-15 on the expression of NKG2D and the cytotoxicity of edited-NK cells against human nasopharyngeal carcinoma cell line CNE2.Methods： NK cells were purified by anti-CD56 MACS and were divided into four groups： non-edited-NK cells group（NK cells treated with 100 U/ml IL-2）,edited-NK cells group（NK cells co-cultured with CNE2 cells at a ratio of 10∶1 and then treated with 100 U/ml IL-2）,edited-NK cells retreated with 1 000 U/ml IL-2 group,and edited-NK cells retreated with 10 ng/ml IL-15 group.Expression of NKG2D in each group was determined by FACS 24 h later.Cytotoxicity of NK cells against CNE2 cells（NK∶CNE2 being 20∶1） was measured by LDH releasing assay.Results： The expression of NKG2D in non-edited-NK cells,edited-NK cells,edited-NK cells retreated with IL-2,and edited-NK cells retreated with IL-15 were（97.63±0.83）%,（53.50±1.25）%,（94.47±1.00）%,and（98.07±0.21）%,respectively.The expression of NKG2D on edited-NK cells retreated with IL-2 or IL-15 was significantly increased than that on edited-NK cells（P〈0.01）.The cytotoxicity of non-edited-NK cells,edited-NK cells,edited-NK cells retreated with IL-2,and edited-NK cells retreated with IL-15 against CNE2 cells were（35.90±3.27）%,（4.70±2.30）%,（31.70 ±3.56）% and（40.18±2.94）%,respectively.The cytotoxicity of edited-NK cells was significantly enhanced after retreated with IL-2 or IL-15（P〈0.01）,with those retreated with IL-15 being stronger than those retreated with IL-2.Conclusion： High dose IL-2 and IL-15 can up-regulate the expression of NKG2D on edited-NK cells and restore their cytotoxicity against CNE2 cells,with the efficacy of IL-15 stronger than that of IL-2.