摘要
目的 观察吲哚胺2,3双加氧酶(IDO)在丁酸钠诱导不成熟树突状细胞(DC)抑制T细胞免疫反应中的作用。方法通过粒细胞-巨噬细胞集落刺激因子(GM—CSF)和白细胞介素(IL)-4体外诱导人外周血来源的DC,第6天加入丁酸钠和不同促成熟因子继续诱导DC分化,逆转录-聚合酶链反应(RT—PCR)、Real-timePCR检测各组IDOmRNA的表达。混合淋巴细胞反应检测各组刺激T细胞增殖能力,进一步在丁酸钠组和对照组分别加入IDO的阻断剂1-甲基色氨酸(1-MT)后了解对T细胞增殖能力的改变。结果RT—PCR结果显示丁酸钠组DCIDOmRNA表达量(0.84±0.01)高于对照组(0.55±0.01)及脂多糖(LPS,0.53±0.01)等诱导成熟组。Real—timePCR进一步显示与对照组比较,丁酸钠组IDOmRNA的表达升高了约(32.03±4.01)倍。MLR显示丁酸钠组DC刺激淋巴细胞增殖指数(1.53±1.0)低于LPS(9.87±3.8)等成熟组DC,并且采用IDO的抑制剂1-MT可以降低抑制T细胞增殖的能力。结论丁酸钠诱导下DC可能通过IDO途径降解细胞微环境中的色氨酸来发挥对T细胞增殖的抑制作用。
Objective To investigate the role of indoleamine 2,3-dioxygense (IDO) in T cell an- ergy of the immature dendritic cells (DCs) induced by sodium butyrate. Methods Human monocyte-derived immature DCs were induced with granuloeyte macrophage-eolony stimulating factor (GM-CSF) and interleukin-4 (IL-4), and divided into 5 groups on the 6th day. Trizol was added to extract total RNA of each group after 24-h induction. IDO mRNA expression was detected by RT-PCR and real-time PCR. 1- methyl-tryptophan (1-MT) was also used to test mixed lymphocyte reaction in sodium butyrate-induced DCs. Cells were harvested after 96-h incubation and fluorescence intensity was detected by FCM. Results IDO mRNA expression in sodium butyrate group was significantly higher (0.84 ±0.01 ) than control group (0.55± 0.01 ) and other mature groups (0.53 ± 0.01 ) by RT-PCR. The result was further confirmed by real-time PCR. In comparison with the control group, the IDO mRNA expression of DCs was increased by (32.03±4.01 ) fold. The proliferation of lymphoeytes was increased in sodium butyrate group with 1-MT by FCM ,which suggested 1-MT may antagonize the inhibition of sodium butyrate-induced lymphocyte proliferation. Conclusion Sodium butyrate may inhibit the lymphocyte proliferation by IDO that can degradate amino-indole propionic acid in microenvironment.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2009年第4期447-449,共3页
Chinese Journal of Experimental Surgery
基金
高等学校博士学科点专项科研基金资助项目(200805581153)
中山大学新教师青年基金资助项目(2007013)