Jagged-1抑制Th17细胞分化的机制初探.docVIP

  Jagged-1 抑制 Th17 细胞分化的机制初探# 王媛，邢飞跃** （暨南大学组织移植与免疫研究中心，广州 510632） 5 10 15 20 25 30 35 40 摘要：为了探讨 Jagged-1 抑制 CD4+T 细胞向 Th17 细胞分化的可能机制，用可溶性 Jagged-1/Fc 嵌合蛋白（ Jagged-1）处理由免疫磁珠分选小鼠淋巴结富集的 CD4+T 细胞，以激活 Jagged-1-Notch 信号通路。通过流式细胞术分析 CD4+ T 细胞中 IL-17A 的表达；Trizol 法提 取细胞总 RNA，利用 RT-PCR 在 mRNA 水平证实基因 Il17a、Il17f 的变化；Western blot 检 测 Jagged-1 处理的 CD4+T 细胞中 RORγt 的表达；ELISA 检测 CD4+T 细胞培养上清中 IL-17A 和 IL-17F 的表达水平。结果显示：Jagged-1 能降低 CD4+ IL-17+ T 细胞的百分比；Jagged-1 组中 Il17a、IL17f mRNA 的表达明显下调。Jagged-1 明显降低 CD4+ T 细胞 RORγt 蛋白水平 的表达；ELISA 结果显示，经 Jagged-1 处理后 CD4+ T 细胞培养上清中 IL-17A，IL-17F 的水 平均降低。结果提示 Jagged-1 能抑制 CD4+ T 细胞向 Th17 细胞分化，其作用与 Jagged-1 信 号下调 IL-17A、IL-17F、RORγt 有关。 关键词：Jagged-1；Th17；分化 中图分类号：Q7,R392 Preliminary mechanism of the differentiation of Th17 cells treated by Jagged-1 WANG Yuan, XING Feiyue (Institute of Tissue Transplantation and Immunology, Jinan University, GuangZhou 510632) Abstract: A soluble Jagged-1/Fc chimera protein (Jagged-1) was directly used to activate Jagged-1-Notch signaling to investigate effect of Jagged-1 on the differentiation CD4+ T cells into Th17 cells. CD4+ T cells were isolated from mouse lymphocytes using magnetic-activated cell sorting (MACS) and treated with Jagged-1. Flow cytometry was performed to analyse the expression of IL-17A on CD4+ T cells. RT-PCR were employed to calculate the mRNA of Il17a and Il17f. Western blot was used to detect RORγt protein level in CD4+ T cells. ELISA was used to determine the level of IL-17A, IL-17F from the culture supernatant of the cultured CD4+ T cells. The results indicated that the soluble Jagged-1/Fc suppressed CD4+ T cells to differentiate into CD4+IL-17+ cells. The mRNA of Il17a and Il17f were significantly inhibited by Jagged-1. Jagged-1 down-regulated the expression of RORγt protein in CD4+ T cells. Jagged-1 suppressed the secretion of IL-17A and IL-17F in the treated cells. The results suggest Jagged-1 inhibits the skewing of CD4+ T cells toward Th17 cells, associated with suppression the