摘要
目的观察白细胞介素-21（IL-21）对人细胞因子诱导的杀伤细胞（CIK）体外培养的影响。方法以常规培养基诱导的CIK细胞为对照组,培养体系中加入IL-21后体外诱导培养10 d,观察细胞增殖能力,并检测CIK细胞表型,分析CIK细胞分泌的干扰素（IFN）-γ、肿瘤坏死因子-α（TNF-α）等细胞因子含量,检测CIK细胞对食管癌细胞株EC9706细胞的毒性作用及凋亡。结果IL-21组细胞数量[（14.58±1.81）×10^-8/L]高于对照组[（7.83±1.18）×10^-8/L,P〈0.05],CD3-＋细胞比率[（98.48±0.28）%]高于对照组[（95.02±1.18）%,P〈0.05],CD3-＋CD4-＋细胞比率[（51.53±1.21）%]高于对照组[（33.36±1.91）%,P〈0.01]。IL-21组CD3-＋CD8-＋细胞比率、CD3-＋CD56-＋虽略高于对照组,但差异无统计学意义（P〈0.05）。IL-21组培养上清液TNF-d浓度为25.0μg/L,高于对照组的12.5μg/L（P〈0.05）,但是两组之间IFN-γ浓度差异无统计学意义（P〈0.05）。IL-21组EC9706细胞的凋亡比率为（15.33±0.91）%,高于对照组的（13.25±1.17）%（P〈0.05）。结论IL-21能增加CIK细胞增殖,提高CD3-＋、CD3-＋CD4-＋阳性表达率,增加细胞因子TNF-α的分泌,促进CIK细胞诱导食管癌细胞凋亡的能力。
Objective To analyze the effect of interleukin （IL）-21 on induction of cytokine-in- duced killer cells （CIK） from human peripheral blood mononuclear cells （PBMCs） in vitro. Methods CIKs were induced from PBMCs of healthy volunteers. The control group was only given conventional medium, and IL-21 was given in IL-21 experimental group. The cells were cultured in an incubator with 5% CO2 at 37 ℃for 10 days. The proliferation of CIK was analyzed by cell counting trypan blue exclusion test. Fluorescence labeled antihuman CD3, CD4, CD8 and CD56 antibodies were used for FCM analysis. The cytokines in the culture medium produced by CIK, including interferon （IFN）--γ, tumor necrosis factor （TNF）-or and IL-12, were analyzed by enzyme-linked immunosorbent assay （ELISA）. Cytotoxcity on e sophageal cancer cells （EC9706） was analyzed by cell counting kit-8 （CCK-8） and apoptosis assays. Re-suits Cell density in IL-21 experimental group was （ 14.58 ± 1.81 ） x 10^5/L, greater than that in control group （7. 83 ± 1.18） x 10^5/L （P 〈 0. 05 ）. In IL-21 experimental group, proportion of CD3 cells was （98.48±0.28）%, higher than that in control group [（95.02 ±1.18）%] （P〈0.05）, and that of CD3 ＋ CIM ＋ was （51.53 ± 1.21 ） %, higher than in control group [ （33.36 ± 1.91 ） % ] （P 〈 0. 05）. No obvious difference in the proportion of CD3 ± CD8 ＋ or CD3 ± CD56 ± was found between two groups. The concentration of TNF-ot in CIK culture medium in IL-21 experimental group was 25.0 μg/L, higher than 12. 5 μg/L in control group （P 〈0. 05）, but there was no significnat difference in IFN-a concentration. Apoptosis rate was （ 15.33 ±0. 91 ） % in IL-21 experimental group, higher than （ 13.25 ±1.17） % in control group （P 〈 0. 05 ）. Conclusion IL-21 can promote the proliferation of CIK. It can increase the pro portion of CD3 ± , and CD3＋CD4 ＋ in CIK. IL-21 can increase the TNF-a production in CIK, promoting the apoptosis of tumor cells
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2014年第7期1524-1526,共3页
Chinese Journal of Experimental Surgery
