IL-1β促进间充质干细胞诱导RA患者CD8^+IL-10^+T细胞形成的作用研究

IL-1β enhances mesenchymal stem cells to induce CD8^+IL-10^+T cell differentiation in patients with rheumatoid arthritis

目的探讨IL-1β在人胎盘间充质干细胞(human placenta-derived mesenchymal stem cells,hPMSCs)调节类风湿关节炎(rheumatoid arthritis,RA)患者活化外周血单个核细胞(peripheral blood mononuclear cells,PBMCs)向CD8^+IL-10^+T细胞亚群分化中的作用。方法应用CBA试剂盒检测RA患者血清IL-1β水平;酶消化法分离健康人hPMSCs;Ficoll密度梯度离心法分离RA患者和健康人PBMCs,分别应用PHA和CD3/CD28单克隆抗体(monoclonal antibody,McAb)活化PBMCs,并与hPMSCs进行共培养。流式细胞术(flow cytometry,FCM)分别分析在程序性死亡配体1(programmed death ligand,PD-L1)和/或PD-L2 McAb阻断及IL-1β刺激条件下hPMSCs对活化PBMCs向CD8^+IL-10^+T细胞亚群分化的诱导能力;应用RT-PCR和FCM分析IL-1β刺激条件下,hPMSCs中PD-L1和PD-L2的表达水平。结果RA患者血清IL-1β水平明显升高。在PHA和CD3/CD28 McAb活化条件下,RA患者PBMCs中CD8^+IL-10^+T细胞亚群比例明显低于健康对照组,且hPMSCs共培养组CD8^+IL-10^+T细胞亚群比例明显升高;IL-1β处理后,hPMSCs对活化PBMCs向CD8^+IL-10^+T细胞亚群分化的诱导能力比未处理组明显增强。IL-1β能够上调hPMSCs对PD-L1和PD-L2的表达;阻断PD-L1和/或PD-L2的表达后,hPMSCs对CD8^+IL-10^+T细胞亚群的诱导分化能力明显减弱。结论IL-1β通过上调hPMSCs对PD-L1和PD-L2的表达,增强hPMSCs对RA患者CD8^+IL-10^+T细胞亚群分化的诱导能力。

Objective To investigate the role of IL-1βin regulating human placenta-derived mesenchymal stem cells(hPMSCs)to induce the differentiation of activated peripheral blood mononuclear cells(PBMCs)into CD8^+IL-10^+T cell subset in patients with rheumatoid arthritis(RA).Methods Serum IL-1βlevels in RA patients were detected by CBA kit.hPMSCs were isolated from healthy subjects by enzyme digestion.PBMCs that were isolated from RA patients and healthy subjects by Ficoll density gradient centrifugation were activated with PHA and CD3/CD28 monoclonal antibody(McAb)and then co-cultured with hPMSCs.Flow cytometry(FCM)was used to analyze the ability of hPMSCs to induce the differentiation of activated PBMCs into CD8^+IL-10^+T cell subset with the presence of programmed death ligand 1(PD-L1)and/or PD-L2 McAb and IL-1β.Expression of PD-L1 and PD-L2 on hPMSCs under IL-1βstimulation was analyzed by RT-PCR and FCM.Results Serum IL-1βlevels were significantly higher in RA patients than in healthy subjects.After the activation by PHA and CD3/CD28 McAb,the proportion of CD8^+IL-10^+T cells in PBMCs of RA patients was significantly lower than that of healthy subjects.Moreover,the proportion of CD8^+IL-10^+T cells significantly increased after co-cultured with hPMSCs.The ability of IL-1β-treated hPMSCs to induce activated PBMCs to differentiate into CD8^+IL-10^+T cells was significantly enhanced compared to untreated hPMSCs.IL-1βcould up-regulate the expression of PD-L1 and PD-L2 on hPMSCs.Blocking the expression of PD-L1 and/or PD-L2 on hPMSCs significantly reduced the ability of hPMSCs to induce activated PBMCs to differentiate into CD8^+IL-10^+T cells.Conclusions IL-1βenhanced the capacity of hPMSCs to induce activated PBMCs to differentiate into CD8^+IL-10^+T cells in RA patients by up-regulating the expression of PD-L1 and PD-L2.