GM-CSF高效诱导人外周血单核细胞向CDl4^＋HLA-DR^-iNOS＋髓系抑制性细胞转化  

Granulocyte-macrophage colony-stimulating factor effectively induces CD14＋ HLA-DR- iNOS＋ myeloid derived suppressor cells from peripheral blood monocytes

作　　者：林哲闽[1] 吉嘉伟 蒋一航 封素娟[1] 谢大炜 张小东[1] 

机构地区：[1]首都医科大学附属北京朝阳医院泌尿外科,北京100020 [2]首都医科大学研究生院

出　　处：《中华器官移植杂志》2017年第7期422-429,共8页Chinese Journal of Organ Transplantation

摘　　要：目的探讨粒-巨噬细胞集落刺激因子（GM-CSF）对于人外周血单核细胞向髓系抑制性细胞（MDscs）转化的影响，进而建立高效的体外诱导体系。方法收集2017年1月至2017年3月肾移植受者的资料，本次实验纳入11例肾移植受者进行研究。术前留取新鲜外周血磁珠分选单核细胞。应用5μg／L和10μg／L等不同浓度GM-CSF处理培养7d，利用流式细胞学分析细胞表型变化，并比较不同组问诱导所得MDsCs（iMDSCs）免疫抑制功能。通过检测iMDSCs诱导型一氧化氮合酶（iN0s）转录水平，明确iNOS途径对于其免疫抑制功能的影响。结果GM-CSF处理后单核细胞HLA-DR水平下降，iMDSCs表型符合单核样MDsCs（M_MDSCs）特征。当iMDSCs／外周血单核细胞（PBMc）为1／2时共培养，可显著抑制T细胞增殖和分泌IL-2和IFN-7能力。检测细胞内iNOS转录水平可见iMDSCs显著高于诱导前单核细胞，加入N0s抑制剂L-单甲基精氨酸（L_NMMA，1mM）后，iMDSCs抑制T细胞增殖作用明显减弱。iMDSCs表型和功能变化与GM-CSF浓度相关，10μg／坳理效果优于5μg／L组。结论应用粒一巨噬细胞集落刺激因子（GM-CSF）处理外周血单核细胞，可高效转化为M-MDSCs，通过iNOS途径显著抑制T淋巴细胞增殖和细胞因子分泌，这一结果有助于建立高效的体外诱导体系，将为MDSCs的临床应用奠定基础与条件。Objective To investigate the effects of granulocyte-macrophage colony-stimulating factor （GM-CSF） on the generation of human myeloid derived suppressor cells （MDSCs） relied on peripheral blood monoeytes, and to establish efficient induction system in vitro of MDSCs. Methods Kidney transplantation recipients between January and March 2017 were included in this study. Purified CD14^＋ cells isolated from peripheral blood were cultured in the presence of GM-CSF with different concentrations for 7 days. Phenotypes and immunosuppressive abilities of induced MDSCs （iMDSCs） were investigated with FACS analyses. Inducible nitric oxide synthase （iNOS） mRNA expression was detected by qRT-PCR to determine the influence of iNOS-pathway on the immunosuppressive abilities of iMDSCs. Results A total of 11 recipients were included in this study. HLA-DR expression decreased sharply after the culture with GM-CSF. iMDSCs showed the similar phenotype characteristics with monoeytic-MDSCs （M-MDSCs） as well as significant ability to suppress T cells proliferation and cytokines productiorL iMDSCs expressed higher levels of iNOS than monocytes, and the inhibitor effects of iMDSCs were significantly reduced after treatment with L-NMMA （I retool/L）. The variations of phenotype and suppressive ability were concentration- dependent, and more significant changes could be revealed in the group of 10 μg/L GM-CSF. Conclusion GM-CSF-treated peripheral blood monocytes can be efficiently transformed to M-MDSCs, and suppress T cells proliferation and cytokines secretion via iNOS-dependent pathway. These results may contribute to establish MDSCs induction system, which would provide a basis for the clinical application of MDSCs.

关 键 词：髓系抑制性细胞 单核细胞 诱导体系 体外 

分 类 号：R699.2[医药卫生—泌尿科学]