摘要
目的:寻求增强CIK细胞的细胞毒活性的有效方法.方法:从健康人外周血中提取出单个核细胞,第1天加入IFN-γ,第2天加入IL-1,CD3 mAb,IL-2诱导CIK细胞,另一组与IL-1.CD3 mAb,IL-2同时加入IL-24.杀伤分为4组:未加IL-24培养组、单独IL-24杀伤组、加IL-24培养组、未加IL-24的CIK细胞培养组在杀伤时加入IL-24.细胞计数法测定细胞的增殖、MTT法测定细胞杀伤活性和流式细胞术分析细胞表型.扫描电镜和透射电镜观察CIK细胞对肿瘤细胞的杀伤和肿瘤细胞的改变.结果:未加IL-24培养组CIK细胞增殖高于加IL-24培养组,两者比较有明显差异(126.34±2.14 vs 108.87±1.29,P<0.05).加IL-24培养组细胞各个效靶比杀伤活性均达到90%以上,明显高于其他各组(效靶比为10:1时95.58%±2.21% vs 27.31%±2.69%,8.74%±2.41%,38.65%±21.30%,P<0.05;效靶比为20:1时91.97%±4.21% vs 34.27%±0.85%,11.54%±2.78%,48.32%±11.72%,P<0.05;效靶比为40:1时91.84%±9.28% vs 50.67%±1.30%,23.73%±11.07%,52.89%±12.26%,P<0.05).不同时间加IL-24培养组各个细胞表型与未加IL-24培养组相比没有差别.透射电镜下观察加IL-24培养组凋亡和坏死肿瘤细胞比未加IL-24培养组明显增多.结论:CIK细胞诱导过程中加入IL-24能明显增强其杀伤活性.
AIM： To find a new method to enhance the cytotoxic activity of cytokine-induced killer （CIK） cells for clinical immunotherapy.
METHODS： Mononuclear cells were extracted from the peripheral blood of health adults. One group was treated with interferon-γ （IFN-γ） at the 1^st day and interleukin-1 （IL-1）, CD3 mAb, IL-2 at the 2^nd day, and the other group was disposed with IL-24 besides IL-1, CD3 mAb, and IL-2. Cell counting method was used to determine the proliferation of CIK cells, and the cytotoxic activity was detected by MTT assay. Cell phenotype was examined by flow cytometry. The scanning electron microscopy and transmission electron microscopy were used to observe the killing effects of CIK cells on tumor cells as well as the changes of tumor cells.
RESULTS： The proliferation of CIK cells without IL-24 treatment was higher than that with IL-24 treatment （126.34 ± 2.14 vs 108.87 ± 1.29, P 〈 0.05）. The cytotoxic activities of CIK cells co-cultured with IL-24 were above 90% at different effectortarget ratios （10 ： 1, 20 ： 1, 40 ： 1）, which were significantly higher than those in the other groups （10 ： 1： 95.58% ± 2.21% vs 27.31% ± 2.69%, 8.74% ± 2.41%, 38.65% ± 21.30%, all P 〈 0.05; 20 ： 1： 91.97%± 4.21% vs 34.27% ± 0.85%, 11.54% ± 2.78%, 48.32% ± 11.72%, all P 〈 0.05; 40 ： 1： 91.84% ± 9.28% vs 50.67% ± 1.30%, 23.73% ± 11.07%, 52.89% ± 12.26%, all P 〈 0.05）. Cell phenotypes were not significantly different between the CIK cells with and without IL-24 treatment （P 〉 0.05）. Apoptotic and necrotic cells were obviously increased in after IL-24 treatment under transmission electron microscope.
CONCLUSION： IL-24 can obviously enhance the cytotoxic activity of CIK cell.
出处
《世界华人消化杂志》
CAS
北大核心
2007年第6期548-553,共6页
World Chinese Journal of Digestology
基金
国家自然科学基金
No.39270765
黑龙江省卫生厅基金
No.2006-478~~
