γ干扰素上调人胎盘间充质干细胞PD-L1表达并增强其诱导脐血和外周血T细胞向IL-10^+T细胞的分化  

IFN-γ up-regulates PD-L1 expression in human placenta mesenchymal stem cells and enhances cell ability to induce the differentiation of IL-10^+T cells from cord blood-and peripheral blood-derived T cells

作　　者：王炜玮[1] 李恒[1] 徐逢皇 都海波 李晓华 易君祝 王国艳[4] 栾希英[1] 

机构地区：[1]滨州医学院免疫学教研室 [2]烟台市莱山区第一人民医院妇产科 [3]烟台市中心血站 [4]滨州医学院烟台附属医院检验科,山东烟台264003

出　　处：《细胞与分子免疫学杂志》2016年第2期191-195,200,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(31370905;31540015)

摘　　要：目的比较人胎盘间充质干细胞(h PMSC)对脐血和外周血IL-10^+T细胞分化的调节作用,并探讨γ干扰素(IFN-γ)在其中的作用机制。方法应用酶消化法分离、培养h PMSC;反转录PCR和流式细胞术分别检测程序性死亡蛋白配体1(PD-L1)在h PMSC的表达;Ficoll密度梯度离心法分离脐血和健康成人外周血单个核细胞,并用红细胞花环法纯化T细胞;用植物血凝素(PHA)活化T细胞并与h PMSC进行共培养,流式细胞术分别检测在阻断型PD-L1 m Ab和IFN-γ预刺激的条件下h PMSC对脐血和外周血T细胞向CD4^+IL-10^+T细胞、CD8^+IL-10^+T细胞分化的诱导作用。结果 h PMSC可诱导脐血和外周血T细胞向CD4^+IL-10^+T细胞、CD8^+IL-10^+T细胞分化,且对外周血T细胞向IL-10^+T细胞分化的诱导能力明显高于对脐血T细胞向该细胞分化的诱导能力;IFN-γ预处理h PMSC后,h PMSC促进脐血和外周血T细胞向IL-10^+T细胞分化能力明显增强;h PMSC高表达PD-L1,IFN-γ可明显上调其在h PMSC上的表达;阻断PD-L1在h PMSC上表达后,脐血、外周血中CD4^+IL-10^+T细胞、CD8^+IL-10^+T细胞比例均明显降低。结论 h PMSC诱导外周血T细胞向IL-10^+T细胞分化的能力明显高于其诱导脐血T细胞向该细胞分化的能力。IFN-γ可通过上调PD-L1在h PMSC上的表达,增强h PMSC对脐血和外周血T细胞向IL-10^+T细胞分化的诱导能力。Objective To compare the differentiation,inducing effects of human placenta mesenchymal stem cells（ h PMSCs） on IL-10^＋T cells derived from cord blood and peripheral blood,and investigate the effect of IFN-γ on the induction. Methods The h PMSCs were isolated from human placenta and cultured. The expression of programmed death ligand 1（ PD-L1） in h PMSCs was detected by reverse transcriptase PCR and flow cytometry（ FCM）, respectively.Mononuclear cel s were isolated from cord blood and peripheral blood of healthy donors by Ficol density gradient centrifugation,and T cells were purified by sheep red blood cells. Then h PMSCs,pretreated with PD-L1 m Ab or IFN-γ,were co-cultured with phytohaemagglutinin（ PHA）-activated T cells. Percentages of CD4^＋IL-10^＋and CD8^＋IL-10^＋T cells in cord blood and peripheral blood T cells were analyzed by FCM. Results h PMSCs could induce the differentiation of CD4^＋IL-10^＋and CD8^＋IL-10^＋T cells from cord blood or peripheral blood T cells,and the number of IL-10^＋T cells in the peripheral blood T cells was significantly higher than that in the cord blood T cells. Pretreatment with IFN-γ markedly enhanced the differentiation,inducing ability of h PMSCs. PD-L1 was highly expressed in h PMSCs,and the expression was also significantly promoted by IFN-γ.After the expression of PD-L1 was blocked in h PMSCs,the percentages of CD4^＋IL-10^＋and CD8^＋IL-10^＋T cells obviously decreased in cord blood and peripheral blood T cells. Conclusion The ability of h PMSCs to induce the differentiation of IL-10^＋T cells from peripheral blood T cells was apparently stronger than that in cord blood T cells. IFN-γ could up-regulate the number of IL-10^＋T cells differentiated from cord blood and peripheral blood T cells in the present of h PMSCs by enhancing the expression of PD-L1 in h PMSCs.

关 键 词：胎盘间充质干细胞 程序性死亡蛋白配体1 IL-10+T细胞 γ干扰素(IFN-γ) 

分 类 号：R392.4[医药卫生—免疫学] R322.12[医药卫生—基础医学]