| 白喜龙,姚含秉,黄斯勇,张 蓉,崔江霞,梁英民,李天晴.IL-17对人脐带间充质干细胞体外免疫调节特性的影响[J].现代生物医学进展英文版,2021,(1):13-20. |
| IL-17对人脐带间充质干细胞体外免疫调节特性的影响 |
| Effect of IL-17 on the Immunoregulatory Characteristics of Human Umbilical Cord Mesenchymal Stem Cells in Vitro |
| Received:May 28, 2020 Revised:June 23, 2020 |
| DOI：10.13241/j.cnki.pmb.2021.01.003 |
| 中文关键词: 人脐带间充质干细胞 干扰素-γ 白介素-17 免疫调节能力 炎症微环境 |
| 英文关键词: Human umbilical cord mesenchymal stem cells IFN-γ IL-17 Immunoregulation capability Inflammatory microenvironment |
| 基金项目:国家重点研究发展计划项目（2018YFA0108502）；陕西省自然科学基础研究计划项目（2019JQ-997）；陕西卫生健康委科研基金项目（2018E004）；西安市科技计划项目（20YXYJ0007(3)） |
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| 中文摘要: |
| 摘要 目的：研究白细胞介素17（IL-17）对人脐带间充质干细胞（hUC-MSCs）的免疫调节能力的影响。方法：采用组织块贴壁法从健康足月胎儿脐带中分离培养hUC-MSCs，并对其进行鉴定。然后在培养基中加入不同浓度IL-17，采用EdU细胞增殖法检测hUC-MSCs增殖情况，流式细胞术（FCM）检测hUC-MSCs表面标记及细胞周期，RT-qPCR检测免疫调节相关基因表达水平，ELISA检测上清中免疫调节相关因子分泌水平，免疫学反应检测其免疫调节能力。结果：①倒置显微镜下观察可见hUC-MSCs呈现较均一漩涡状、梭形生长，细胞体积较小；细胞表面表达CD44、CD73、CD90、CD105，不表达CD34、CD19、CD11b、CD45、HLA-DR；具有成脂、成骨、成软骨诱导分化能力。②FCM检测结果显示IL-17处理后，hUC-MSCs免疫表型未发生变化，而IFN-γ处理后，hUC-MSCs表面标记HLA-DR表达明显上调。③与对照组相比，IL-17处理组的hUC-MSCs活性更好，且能促进细胞增殖。④与空白对照组相比，IL-17处理组hUC-MSCs的TGF-β、IL-10、IDO、PGE2、TSG-6、PD-L1基因表达均显著上调（P<0.001）。⑤ELISA检测结果显示，与空白对照组相比，IL-17处理组上清中TGF-β、IL-10、IDO、PGE-2、TSG-6、PD-L1分泌水平明显增高（P<0.001）。⑥与空白对照组相比，IL-17处理组的CD32CD42T细胞、CD32CD82T细胞、B细胞的比例降低，Treg细胞比例增高（P<0.001）。结论：IL-17在不影响hUC-MSCs免疫表型的条件下，可促进细胞增殖，上调免疫相关基因表达，提高TGF-β、IL-10、PGE-2、IDO、TSG-6、PD-L1分泌水平，增强hUC-MSC免疫调节能力。因此，本研究将为预处理的hUC-MSCs防治免疫炎症性疾病提供理论依据。 |
| 英文摘要: |
| ABSTRACT Objective: To observe the effects of interleukin 17(IL-17) on the immunomodulatory capacity of human umbilical cord mesenchymal stem cells (hUC-MSCs). Methods: hUC-MSCs were isolated from the umbilical cord of healthy term fetus by tissue explants adherent method, and cultured with serum-free medium with IL-17. EdU cell proliferation method was used to detect the proliferation of hUC-MSCs, Flow cytometry was used to detect cell surface markers and cell cycle, gene expression levels related to immune regulation was detected by RT-qPCR, the secretion level of immunomodulatory factors was detected by ELISA, and the immunomodulatory ability of hUC-MSCs was detected by immunological experiment. Results: ①Under the inverted microscope, hUC-MSCs cultured with with serum-free medium showed more uniform vortex-like and spindle growth. hUC-MSCs were expressed CD44, CD73, CD90 and CD105, but lowly expressed CD34, CD19, CD11b, CD45, and HLA-DR. Results from the induced differentiation experiments showed that hUC-MSCs had adipogenic, osteogenic and chondrogenic abilities. ②After IL-17 pre-treatment, the immunophenotype of hUC-MSCs was not changed, however, after IFN-γ pre-treatment, the expression of HLA-DR on hUC-MSCs was significantly up-regulated. ③As compared with control group, the cell viability and cell proliferation of hUC-MSCs were pre-treatment by IL-17 was better. ④Compared with the blank control group, the gene expressions of TGF-β, IL-10, IDO, PGE2, TSG-6, and PD-L1 of hUC-MSCs were pre-treatment by IL-17 were significantly up-regulated(P<0.001). ⑤Compared with the blank control group, the secretion levels of TGF-β, IL-10, IDO, PGE-2, TSG-6 and PD-L1 were significantly increased in the supernatant of UC-MSCs were pre-treatment by IL-17 group (P<0.001). ⑥hUC-MSCs were pre-treatment by IL-17 can reduce the proportion of CD3+CD4+T cells, CD3+CD8+Tcells, and B cells in lymphocytes, but increase the proportion of Treg cells(P<0.001). Conclusion: Without affecting the immunophenotype of hUC-MSCs, IL-17 can promote cell proliferation, up-regulate the expression of immune-related genes, increase the secretion level of TGF-β, IL-10, PGE-2, IDO, TSG-6, PD-L1, and enhance immune regulation ability of hUC-MSC. Therefore, this study will provide a theoretical basis for priming hUC-MSCs to prevent and treat immune inflammatory diseases. |
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