MSCs通过促分泌TGF-β1、IL-6和IL-10抑制干燥综合征患者CD4+T细胞的活化增殖

Mesenchymal stem cells inhibit activation and proliferation of CD4+ cells in Sjgren syndrome by promoting TGF-β1, IL-6 and IL-10 secretion

关键词:  干燥综合征  脐带间充质干细胞  TGF-β1  IL-6  IL-10  免疫抑制效应

Key words:  Sj gren syndrome  umbilical cord mesenchymal stem cells  TGF β1  IL 6  IL 10  immunosuppressive effects

摘要:

目的 体外探讨人脐带间充质干细胞(mesenchymal stem cells, MSCs)对原发性干燥综合征(primary Sjgren syndrome, pSS)患者外周血活化CD4⁺ T细胞的免疫抑制效应,分析MSCs促分泌的转化生长因子-β1(transforming growth factor-β1, TGF-β1)、IL-6及IL-10在免疫抑制效应中的作用。方法 原代分离脐带MSCs,并经流式细胞术鉴定。分选成年健康者和pSS患者外周血CD4⁺T细胞,分为对照组活化CD4⁺T细胞(CD3、CD28抗体共刺激72h)、MSCs共培养组(活化CD4⁺T细胞与MSCs共培养72h)和干扰素-γ(IFN-γ)预刺激后MSCs共培养组(活化CD4⁺T细胞与IFN-γ预刺激后的MSCs共培养72h),活化CD4⁺T细胞组为对照。ELISA法检测培养上清液中TGF-β1、IL-6、IL-10因子水平。结果 相对于活化CD4⁺T细胞组,MSCs共培养组培养上清中TGF-β1、IL-6、IL-10浓度均显著升高(P≤0.01),并且活化的CD4⁺T细胞增殖受到抑制。结论 MSCs通过促分泌TGF-β1、IL-6、IL-10抑制活化的CD4⁺T细胞增殖,起到免疫抑制作用。

Abstract:

Objective To investigate the immune suppression effect of human umbilical cord mesenchymal stem cells (MSCs) on the activation of peripheral blood CD4⁺ T cells in primary Sjgren syndrome (pSS) in vitro, and analyze the immunosuppressive effects of transforming growth factor-β1 (TGF-β1 ), IL-6 and IL-10 promoted by MSCs. Methods Primary cultured umbilical cord MSCs were identified by flow cytometry (FCM),and peripheral blood CD4⁺T cells were sorted both in healthy adults and in pSS patient. CD4⁺T cells were cultured with CD3, CD28 antibody for 72h to be activated(control group)； the activated CD4⁺T cells were co-cultured with MSCs for 72h(MSCs group)or MSC were pre-stimulated with interferon-γ (IFN-γ), then the activated CD4⁺T cells were co-cultured with pre-stimulated MSCs for 72h(pre-stimulated group). TGF-β1, IL-6 and IL-10 in the supernatants were detected by ELISA.Results Compared with control group, levels of TGF-β1, IL-6 and IL-10 were significantly increased in MSCs group and pre-stimulated MSCs group(P≤0.01), and CD4⁺T cell activation was inhibited. Conclusion MSCs may have an immunosuppressive effect,which can inhibit the proliferation of activated CD4⁺T cells by promoting the secretion of TGF-β1, IL-6 and IL-10.