CAJ | 학술논문

目的探讨表达吲哚胺2,3-二氧化酶(IDO)的Kupffer细胞(KC)在体外对小鼠同种异体T淋巴细胞(TC)增殖的抑制作用.方法用实时荧光定量PCR检测经IFN-γ处理或未处理的KC表达IDOmRNA和FasLmRNA的情况.用高压液相色谱仪分析IDO对色氨酸的分解作用以了解其活性.用3H嵌入增殖试验检测KC对同种异体TC增殖抑制情况.用流式细胞仪分析KC对同种异体TC细胞周期的影响和凋亡作用.结果实时荧光定量PCR显示:经IFN-γ处理后小鼠KC表达IDO和FasL并且其表达在6 h后均达到高峰.IDO能降低培养基中色氨酸的浓度,提高其代谢产物犬尿氨酸浓度.表达IDO的KC能明显抑制同种异体TC的增殖,但1-甲基色氨酸和抗FasL抗体能部分阻断其增殖抑制作用,且存在剂量依赖关系.表达IDO和FasL的KC能阻滞同种异体TC于G1中期,诱导其凋亡.结论除了FasL/Fas途径,IDO可能是KC抑制同种异体TC的增殖并诱导其凋亡的另一机制.
목적 탐토표체신타알2,3-이양화매(IDO)적Kupffer세포(KC)재체외대소서동충이체T림파세포(TC)증식적억제작용.방법 용실시형광정량PCR검측경IFN-γ처리혹미처리적KC표체IDOmRNA화FasLmRNA적정황.용고압액상색보의분석IDO대색안산적분해작용이료해기활성.용3H감입증식시험검측KC대동충이체TC증식억제정황.용류식세포의분석KC대동충이체TC세포주기적영향화조망작용.결과 실시형광정량PCR현시:경IFN-γ처리후소서KC표체IDO화FasL병차기표체재6 h후균체도고봉.IDO능강저배양기중색안산적농도,제고기대사산물견뇨안산농도.표체IDO적KC능명현억제동충이체TC적증식,단1-갑기색안산화항FasL항체능부분조단기증식억제작용,차존재제량의뢰관계.표체IDO화FasL적KC능조체동충이체TC우G1중기,유도기조망.결론 제료FasL/Fas도경,IDO가능시KC억제동충이체TC적증식병유도기조망적령일궤제.
Objective To investigate kupffer cells (KCs) expressing indoleamine 2,3-dioxygenase(IDO)in the inhibition of allogeneic T-cell proliferation in vitro. Methods Real-time PCR was used to investigate the expression of IDO mRNA and FasL mRNA in KCs pretreated with or without IFNγ. High performance liquid chromatography was used to analyze the catabolism of tryptophan by IDO from KCs. Allogeneic T-cell response was used to confirm the inhibition of KCs in vitro. The proliferation of lymphocytes was detected using [3 H] thymidine incorporation. Cell cycle and lymphocyte apoptosis were evaluated by flow cytometric assay. Results Real-time PCR revealed IDO mRNA and FasL mRNA expression in KCs pretreated with IFN-γ. IDO catabolic effect was confirmed by a decrease in tryptophan and increase in kynurenine concentration. KCs expressing IDO and FasL from BABL/c mice acquire the ability to suppress the proliferation of T-cells from C57BL/6, which could be blocked by the addition of 1-methyl-tryptophan and anti-FasL antibody. The co-cultured T-cells with KCs expressing IDO and FasL could induce allogeneic T-cell apoptosis and exhibited cell-cycle arrest in G1. Conclusion In addition to the Fas/FasL pathway, IDO may also play an important role in KCs to inhibit allogeneic T-cell proliferation in vitro.