人羊膜间充质干细胞抑制异体淋巴细胞增殖并减少γ干扰素的分泌  

Human amniotic mesenchymal stem cells inhibit allogeneic lymphocyte proliferation and reduce the secretion of interferon γ

作　　者：宋瑾[1] 丛姗[1] 李岩[1] 白立恒 曹贵方[1] 

机构地区：[1]内蒙古农业大学兽医学院动物组织胚胎学与发育生物学实验室,内蒙古呼和浩特010018 [2]内蒙古妇幼保健医院妇产科,内蒙古呼和浩特010020

出　　处：《细胞与分子免疫学杂志》2015年第3期333-337,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家高技术研究发展计划(863)(2008AA101005)

摘　　要：目的观察人羊膜间充质干细胞(hAMSC)对体外培养的淋巴细胞功能的影响。方法通过酶消化法分离培养hAMSC,采用荧光团标记的小鼠抗人单克隆抗体结合流式细胞术鉴定细胞表面抗原;免疫荧光染色检测培养细胞波形蛋白(vimentin)和阶段特异表达抗原4(SSEA-4)的表达;分离培养hAMSC和外周血单个核细胞(PBMC),将刀豆蛋白(ConA)刺激的淋巴细胞与1×104、5×104、1×105个hAMSC进行共培养。CCK-8法测定淋巴细胞增殖,ELISA测定细胞上清液中IFN-γ的水平。结果5μg/mLConA能够引起淋巴细胞增殖;共培养条件下,hAMSC能够抑制ConA引起的淋巴细胞增殖,且随着hAMSC的数量增加,抑制效果更明显。培养72h后,CCK-8法结果表明,单纯ConA刺激后淋巴细胞数显著高于共培养细胞。选择抑制效果最佳的组别1×106个淋巴细胞与1×105个hAMSC共培养,ELISA测定1×105个hAMSC对淋巴细胞抑制72h后,上清液中IFN-γ分泌,共培养细胞上清液中IFN-γ水平显著低于单纯ConA刺激细胞。结论hAMSC能在体外抑制ConA引起的淋巴细胞增殖并减少IFN-γ分泌。Objective To investigate the effects of human amniotic mesenchymal stem cells （hAMSCs） on the function of lymphocytes in vitro. Methods Enzymatic digestion method was used to isolate and culture hAMSCs. Fluorophore-labeled mouse anti-human monoclonal antibodies were used to identify cell surface antigens with flow cytometry. The expressions of vimentin and stage specific embryonic antigen-4 （SSEA-4） were detected by immunofluorescence staining. Isolated lymphocytes were stimulated by concanavalin （ConA）, and then 1 × 10^4, 5 × 10^4, 1 × 10^5 hAMSCs were co-cultured with the ConA-treated lymphocytes. Lymphocyte proliferation was measured by CCK-8 assay and the supernatant level of IFN-γ was determined by ELISA. Results ConA （5 μg/mL） could cause lymphocyte proliferation, hAMSCs inhibited lymphocyte proliferation induced by ConA in co-culture conditions, and with the increasing number of hAMSCs, the suppressing effect was more obvious. When the cells were cultured for 72 hours, CCK-8 assay showed that the number of lymphocytes treated with ConA alone was significantly higher than that of ConA-treated lymphocytes co-cultured with hAMSCs. The best inhibitory group 1 × 10^6 lymphocytes co-cultured with 1 × 10^5 hAMSCs was selected to measure supematant IFN-γ secretion by ELISA after 72 hours. The level of IFN-γ was significantly lower than that in the simple ConA-stimulated group. Conclusion HAMSCs could inhibit lymphocyte proliferation and reduce IFN-γ secretion induced by ConA in vitro.

关 键 词：人羊膜间充质干细胞 淋巴细胞 干扰素-Γ 免疫抑制 

分 类 号：R392.11[医药卫生—免疫学] R446.61[医药卫生—基础医学]