体外分化Th17细胞IL-17f,ctBP和Gfi1的表达

The Expression of IL-17f,ctBP,Gfi1 in Th17 Cells Differentiated in Vitro

目的检测小鼠Th17细胞诱导分化模型中IL-17f,ctBP和Gfi1的表达情况并探讨其临床意义。方法提取小鼠脾淋巴细胞,采用免疫磁珠纯化CD4+CD62L+T细胞后,空白对照组用RPMI 1640培养,Th17分化组用anti-CD3ε,anti-CD28,anti-IFN-γ,anti-IL-4抗体及IL-6,TGF-β1和IL-23等细胞因子培育,诱导原始T细胞向Th17细胞分化。两组细胞分别培养72 h后,采用Realtime-PCR检测IL-17f,ctBP,Gfi1 mRNA的表达情况。结果 Th17分化组细胞经相关抗体及细胞因子刺激诱导分化后,其IL-17f mRNA表达明显增高(分化前为1,分化后达194.011721),但ctBPmRNA,Gfi1 mRNA表达均明显低于空白对照组,以上差异均有统计学意义(P均<0.01),对照组ctBPmRNA分化前为1,Th17分化后为0.45375958,Gfi1 mRNA表达在对照组分化前为1,Th17分化后为0.00430432。结论 ctBP和Gfi1在Th17分化组中表达显著下调,但上述两种转录辅抑制因子是否参与了Th17细胞的分化调控值得进一步研究。

Objective To determine the expression of IL-17 f, ctBP and Gill in differentiated mouse Thl7 cells and their clinical significance. Methods Mouse spleen mononuclear lymphocytes were purified by mouse CD4 ^＋ CD62L ^＋T Cell isolation Kit II. The CD4 ^＋ CD62L^＋T Cells were divided into control group and Thl7 group. Control group cells were cultured with RPMI 1640 while the differentiated Thl7 Cell group were cultured with anti - mouse CD28, IL-6, TGF-131, anti-mouse IL-4, anti - mouse IFN-γ, and mouse IL-23 at 37℃, 5% CO2 for 72 hours. The expression levels of IL-17f, ctBP, Gfil mRNA were evaluated with Real-time quantitative PCR. Results After stimulated by antibodies and cytokines,the expression levels of IL-17f mRNA in differen- tiated Thl7 cells were significantly increased obviously （1 for pre-differentiation vs. 194.011721 for post-differentiation, P 〈0.01）. In contrast,the expression levels of ctBP and Gill mRNA were downregulated（ 1 for both ctBP and Gill in controls vs. 0.45375958 for ctBP and 0.00430432 Gill in differentiated Thl7 cells, P 〈 0. 01 for both）. Conclusion The down-regulation of CtBP and Gill mRNA expression in differentiated Thl7 cells suggests that these two inhibitory transcription factors are involved in Thl7 cell differentiation.