IL-12与IL-18联合刺激对树突状细胞分泌的exosome活性的影响  

Effect of stimulation with combined IL-12 and IL-18 on exosomes secreted by dendritic cells

作　　者：张在云[1] 李希德 魏玮[1] 叶兰[1] 丛萍[1] 潘祥林[3] 

机构地区：[1]山东大学第二医院肿瘤中心,山东济南250033 [2]高密市中医院肿瘤科,山东高密261500 [3]山东大学第二医院血液内科,山东济南250033

出　　处：《中国肿瘤生物治疗杂志》2012年第1期17-21,共5页Chinese Journal of Cancer Biotherapy

基　　金：山东省自然科学基金资助项目(No.Y2007C111)~~

摘　　要：目的:观察IL-12与IL-18联合刺激对树突状细胞(dendritic cell,DC)分泌的exosome(DC derived exosome,Dex)活性的影响,为探索高效的exosome肿瘤疫苗奠定基础。方法:取正常健康人外周血单个核细胞诱导培养DCs,分别以IL-12、IL-18或IL-12+IL-18联合刺激DC,并设空白对照组、T细胞对照组,分别提取各组的Dex,Western blotting检测Dex中HLA-DR、CD83的表达,流式细胞仪检测CD54、CD80及CD86的表达;MTT法检测Dex刺激T细胞增殖的作用,ELISA法测定T细胞IFN-γ的分泌量。结果:IL-12、IL-18、联合组、空白组的Dex中均有HLA-DR、CD83蛋白表达;联合组Dex的CD54(323.67±44.06 vs 246.17±31.91、236.33±33.87、167.67±28.73,P<0.05)、CD80(406.37±39.18 vs 331.67±36.15、335.67±41.38、260.00±35.58,P<0.05)及CD86(390.50±38.06 vs 314.33±36.64、319.00±33.10、246.83±30.55,P<0.05)表达均高于IL-12组、IL-18组及空白组;联合组刺激T细胞增殖高于IL-12组、IL-18组及空白组、T细胞组(1.98±0.31vs 1.55±0.23、1.57±0.21、1.10±0.18、0.53±0.09,P<0.05);联合组刺激T细胞分泌IFN-γ水平高于IL-12组、IL-18组及空白组、T细胞组(436.67±61.80 vs 295.04±40.25、358.18±55.77、225.00±36.44、139.50±17.63,P<0.05)。结论:IL-12与IL-18联合刺激能增加Dex表达CD54、CD80及CD86,促进Dex刺激的T细胞的增殖及其IFN-γ的分泌。Objective： To observe the effect of stimulation with combined IL-12 and IL-18 on exosomes secreted by dendritic cells（DCs）,and so as to facilitate exploring high-efficiency DC derived exosome（Dex） cancer vaccines.Methods： DCs induced from human peripheral blood mononuclear cells（PBMCs） were stimulated with IL-12＋IL-18,IL-12 or IL-18 for the combined group,IL-12 group and IL-18 group respectively;and no stimulant for the control group.Dex was extracted from DCs.The expressions of HLA-DR and CD83 in Dex were detected by Western blotting.CD54,CD80 and CD86 expressions were determined by flow cytometry.The T cell proliferation stimulated by Dex was detected with MTT method.IFN-γ in the T cell supernatant was detected by ELISA.Results： HLA-DR and CD83 proteins were expressed in Dex of all the four groups.The molecule expressions for combined group were CD54（323.67±44.06 vs 246.17±31.91,236.33±33.87,167.67±28.73,P0.05）,CD80（406.37±39.18 vs 331.67±36.15,335.67±41.38,260.00±35.58,P0.05）,and CD86（390.50±38.06 vs 314.33±36.64,319.00±33.10,246.83±30.55,P0.05）,which were higher than those in the IL-12 group,IL-18 group and control group.The T cell proliferation in the combined group（1.98±0.31 vs 1.55±0.23,1.57±0.21,1.10±0.18,P0.05） was higher than that in the IL-12 group,IL-18 group and control group.IFN-γ secreted by T cells in the combined group（436.67±61.8 vs 295.04±40.25,358.18±55.77,225.00±36.44,P0.05） was higher than that in the IL-12 group,IL-18 group and control group.Conclusion： Stimulation of DCs with combined IL-12 and IL-18 can enhance Dex express CD54,CD80 and CD86,promote Dex-induced T cell proliferation and increase IFN-γ secretion by T cells.

关 键 词：树突状细胞 EXOSOME IL-12 IL-18 T细胞 IFN-Γ 

分 类 号：R392.1[医药卫生—免疫学] R730.51[医药卫生—基础医学]