摘要
目的:探讨IL-12对细胞因子诱导的杀伤(cytokine-induced killer,CIK)细胞的表型和CIK细胞在体外对食管癌EC9706细胞杀伤活性的影响。方法:体外分离人外周血单个核细胞,分为两组:对照组以IFN-γ、IL-2和CD3单抗诱导培养CIK细胞;IL-12组在对照组基础上加用IL-12培养。培养至第14天,流式细胞仪检测两组CIK细胞的免疫表型;LDH释放法测定效靶比20∶1、30∶1时两组CIK细胞对EC9706细胞的杀伤活性;观察效靶比30∶1时,NKG2D单抗对两组CIK细胞杀伤活性的影响。结果:IL-12组与对照组相比较,CIK细胞免疫表型的CD3+CD56+细胞比例[(28.23±1.71)%vs(16.34±0.59)%,P<0.05]、CD3+细胞及CD3+CD56+细胞中NKG2D的表达[(77.45±2.15)%vs(66.87±0.73)%,(92.94±0.77)%vs(82.18±0.66)%;均P<0.05]、CD3+细胞中穿孔素的表达[(51.78±0.63)%vs(43.54±0.95)%,P<0.05]都明显增强;CD3+细胞颗粒酶B表达无明显变化[(26.90±0.67)%vs(26.76±0.33)%,P>0.05)]。效靶比20∶1和30∶1时,IL-12组CIK细胞对EC9706细胞的杀伤活性均较对照组明显增强[(43.92±1.67)%vs(35.34±1.22)%,(55.95±0.88)%vs(43.91±1.10)%;均P<0.05];以NKG2D单抗阻断后,IL-12组、对照组CIK细胞杀伤活性均明显下降[(19.72±0.56)%vs(55.95±0.88)%,(19.83±1.20)%vs(43.91±1.10)%;均P<0.05]。结论:IL-12能够上调CIK细胞NKG2D和穿孔素的表达,从而增强CIK细胞对EC9706细胞的杀伤活性。
Objective： To explore the effects of IL-12 on the phenotypes of cytokine-induced killer （CIK） cells and the cytolytic activity of CIK cells against human esophageal carcinoma EC9706 cells in vitro. Methods： Peripheral blood mononuclear cells were isolated from healthy donors. Two groups were designed ： a control group （ cells were cultured in the presence of IFN-У/, IL-2 and anti-CD3 antibody） and IL-12 group （cells were cultured in the presence of IFN-У/, anti- CD3 antibody , IL-2 and IL-12 ）. After 14-day culture, the phenotypes of CIK cells in the control and IL-12 groups were analyzed by flow cytometry. The cytotoxic activity of CIK cells on EC9706 cells was measured by LDH releasing assay at effect-to-target （ E： T） cell ratios of 20： 1, 30： 1. The anti-NKG2D monoclonal antibody was added to CIK cells to detect its effect on the eytotoxic activity of CIK cells at E： T ratio of 30： 1. Results： In comparison to the control group, the pro- portion of CD3 ＋ CD56 ＋ cells, NKG2D expressions on CD3 ＋ and CD3 ＋ CD56 ＋ cells, and perforin expression in CD3＋ cells were higher in the IL-12 group （ [ 28.23 ± 1.71 ] % vs [ 16.34 ± 0.59 ] % ; [ 77.45± 2.15 ] % vs [ 66.87 ± 0.73 ] %, [92.94±0.77]% vs [82.18 ±0.661%; [51.78 ±0.63]% vs [43.54 ±0.951%; allP〈0.05）. Whereas, no sig- nificant change was observed in the granzyme B expression in the CD3 ~ cells [ （26.90 ± 0.67 ） % vs （26.76 ±0. 33 ） %, P 〉 0.05 ]. The cytolytic activity of CIK cells against EC9706 cells was increased significantly in the IL-12 group （E： T ratio of 20： l , [43.92± 1.671% vs [35.34±1.221% ;E：Tratioof30：l, [55.95 ±0.88]% vs [43.91 ±1.101%, all P 〈 0.05 ）. The cytotoxicity of CIK ceils in the IL-12 and the control groups were significantly inhibited by anti-NKG2D monoclonal antibody （ [ 19.72 ± 0.56 ] % vs [ 55.95 ± O. 88 ] %, [ 19.83 ± 1.20 ] % vs [ 43.91 ± 1.10 ] %, all P 〈 0. 05 ）. Conclusion： IL-12 up-regulates the NKG2D and perforin expressions
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2013年第2期197-200,共4页
Chinese Journal of Cancer Biotherapy
基金
河南省科技计划资助项目(No.201143)~~
