摘要
目的:研究人胰腺癌微环境对树突状细胞(DCs)T细胞免疫球蛋白及黏蛋白-3(TIM-3)表达及其功能的影响,初步探讨肿瘤微环境调节DCs上TIM-3表达的可能机制。方法:流式细胞术检测肿瘤浸润树突状细胞(TIDC)以及癌旁组织、胰腺癌患者和健康人外周血诱导DCs上TIM-3的表达;观察人胰腺癌细胞培养液上清对健康人外周血单个核细胞经rh GM-CSF和rh IL-4诱导扩增制备DCs上TIM-3表达的影响;酶联免疫吸附法(ELISA)检测TIM-3^+DCs组和对照组的DCs分别与凋亡胰腺癌细胞Capan-2共培养上清中细胞因子IFN-β和IL-12水平。结果:胰腺癌组织中TIDC上TIM-3的表达明显高于癌旁组织及患者和健康人外周血的DCs(P<0.01)。人胰腺癌细胞株Canpan-2、SW1990和Panc-1的上清液较人皮肤成纤维细胞Hs27显著上调DCs上TIM-3表达(P<0.05),联合阻断VEGF、IL-10和PGE_2可明显降低Canpan-2细胞上清对DCs上TIM-3的上调作用(P<0.05)。TIM-3高表达DCs组较低表达组分泌的IL-12和IFN-β水平低,而阻断TIM-3后,IFN-β和IL-12水平均升高(P<0.01)。而这种升高趋势可在加入DNase和RNase后消失。结论:人胰腺癌TIDC上TIM-3表达升高导致其固有免疫功能受损;肿瘤细胞分泌的VEGF、IL-10和PGE2可能参与TIM-3的表达调控。
AIM： To investigate the influence and mechanisms of human pancreatic cancer tumor microenvironments on T-cell immunoglobulin mucin-3（ TIM-3） expression and function of dendritic cells（ DCs）. METHODS：Tumor-infiltrating dendritic cells（ TIDC） and para-carcinoma tissue DCs were isolated by Ficoll-Hypaque density centrifugation from trypsinized pancreatic carcinoma tissues,and the peripheral blood mononuclear cells were isolated from pancreatic cancer patients or healthy people. The expression of TIM-3 on DCs was detected by flow cytometry. DCs isolated from healthy people peripheral blood mononuclear cells were induced by rh GM-CSF and IL-4. The expression of TIM-3 in the DCs treated with the culture supernatants of Capan-2,SW1990 and Panc-1 pancreatic cancer cells or human skin fibroblast（ Hs27） cells for 48 h,and in the DCs treated with supernatants of Capan-2 cells,anti-VEGF-R2,anti-IL-10 and EP2 receptor blocking peptide were evaluated by flow cytometry. The releases of IFN-β and IL-12 in the culture supernatants of DCs pretreated with monoclonal antibody（ m Ab） to TIM-3 or DNase ＋ RNase,followed by stimulation with apoptotic Capan-2 cells,were detected by ELISA. RESULTS： DCs in tumor microenvironments had higher expression of TIM-3 than the DCs from para-carcinoma tissues and pancreatic cancer patient or healthy people peripheral blood（ P 0. 01）. TIM-3expression in the DCs treated with the culture supernatants of Capan-2,SW1990 and Panc-1 pancreatic cancer cells for 48 h was much higher than that in Hs27 cells（ P 0. 05）. Treatment with a combination of anti-VEGF-R2,anti-IL-10 and EP2 receptor blocking peptide largely diminished the upregulation of TIM-3 on the DCs mediated by Capan-2 cell supernatants（ P 0. 05）. The concentrations of IFN-β and IL-12 in the DCs with high expression level of TIM-3 were lower than those in the DCs with low TIM-3 expression level. Treatment with m Ab to TIM-3 resulted in much more IFN-β and IL-12releases（ P 0. 01）,but DNase ＋ RNase made this effect disappear. CONCLUSION： TIM-3 serves as a negative regulator of DCs innate immune responses in the pancreatic cancer microenvironments. The secretion of soluble factors to tumor microenvironment by pancreatic cancer cells,including IL-10,VEGF and PGE_2,may contribute to the regulation of TIM-3 expression.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2016年第4期628-636,共9页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81000917
No.81370059)
