胸腺肽α1调节AML细胞株IDO表达对T细胞免疫抑制作用影响的研究

发布时间：2018-04-25 19:17

本文选题：胸腺肽α + 急性髓系白血病　；参考：《中华肿瘤防治杂志》2017年02期

【摘要】：目的吲哚胺2,3-双加氧酶(indoleamine 2,3-dioxygenase,IDO)是色氨酸代谢过程中的限速酶,近年来研究发现,IDO高表达是急性髓系白血病(acute myeloid leukemia,AML)细胞介导免疫耐受的重要机制,因此研究如何调控IDO表达,将有可能为AML治疗提供新的辅助方法。本实验拟从分子生物及蛋白质水平,探讨胸腺肽α1(thymosin-α1,Tα1)调节AML细胞株IDO表达及改善T细胞的免疫抑制作用,为AML免疫治疗提供理论依据。方法采用逆转录-聚合酶联反应(RT-PCR)检测γ-干扰素(interferon-γ,IFN-γ)、Tα1对HL-60/K562细胞株IDO mRNA表达的影响;采用蛋白质印迹法检测IFN-γ和Tα1对HL-60/K562细胞株IDO蛋白表达的影响;采用MTT检测Tα1和1-甲基色氨酸(1-methyl-tryptophan,1-MT)作用后经IFN-γ诱导的HL-60/K562细胞对植物血凝素(phytohaemagglutinin,PHA)刺激T细胞增殖抑制作用。结果HL-60/K562细胞株均表达IDO mRNA;IFN-γ作用后,IDO mRNA表达明显增加(HL-60细胞株t=130.19,K562细胞株t=67.38,均P0.05);IFN-γ+Tα1共同作用后,明显下调IDO mRNA表达(HL-60细胞株F=132.55,K562细胞株F=77.36,均P0.05),并呈浓度依赖性(HL-60细胞株r=-0.71,K562细胞株r=-0.80);HL-60/K562细胞株低水平表达IDO蛋白;经IFN-γ作用后,HL-60细胞株IDO蛋白表达水平为2.505 6±0.019 6,明显高于空白对照组0.189 7±0.083 4,t=326.79,P0.05;K562细胞株IDO蛋白表达水平为0.988 8±0.023 9,明显高于空白对照组0.253 6±0.016 3,t=76.12,P0.05;IFN-γ+Tα1共同作用后,经IFN-γ+Tα1共同作用后,IDO蛋白在HL-60细胞株各组的表达水平分别为2.455 0±0.066 1、1.526 0±0.017 3、1.134 1±0.060 1、0.806 0±0.029 7和0.070 0±0.003 6,并呈浓度依赖性,F=4 187.76,r=-0.77,P0.05;经IFN-γ+Tα1共同作用后,IDO蛋白在K562细胞株各组的表达水平分别为0.998 6±0.016 4、0.784 3±0.014 9、0.714 5±0.009 5、0.656 6±0.007 6和0.492 2±0.010 9,并呈浓度依赖性,F=59.50,r=-0.76,P0.05。IFN-γ诱导的HL-60/K562细胞可抑制PHA刺激的T细胞增殖(P0.05),并呈浓度依赖性(HL-60细胞株r=0.99,K562细胞株r=0.98);1-MT组和Tα1组中HL-60/K562细胞株对T细胞增殖的抑制率明显低于对照组,差异有统计学意义,P0.05。结论 IDO可能在AML诱导机体产生免疫耐受过程中起重要作用,Tα1可下调AML细胞IDO表达,改善AML细胞对T细胞的免疫抑制作用。
[Abstract]:Objective Indoleamine 2ndoleamine 23-dioxygenase (IDO) is a rate-limiting enzyme in tryptophan metabolism. In recent years, it has been found that the high expression of indoleamine-3-dioxygenase (IDO) is an important mechanism of immune tolerance mediated by acute myeloid leukemiaMLs in acute myeloid leukemia (AML) cells, so we studied how to regulate the expression of IDO. It will be possible to provide new adjuvant methods for AML therapy. The aim of this study was to investigate the effects of thymosin- 伪 1T 伪 1) on regulating the expression of IDO in AML cell line and improving the immunosuppressive effect of T cell line from the molecular biological and protein levels, and to provide a theoretical basis for the immunotherapy of AML. Methods reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the effect of interferon- 纬 -interferon- 纬 on the expression of IDO mRNA in HL-60/K562 cell line, and the effect of IFN- 纬 and T 伪 1 on the expression of IDO protein in HL-60/K562 cell line was detected by Western blotting. MTT was used to detect the inhibitory effect of HL-60/K562 cells induced by IFN- 纬 on the proliferation of T cells stimulated by phytohaemagglutinin (PHAs). Results all HL-60/K562 cell lines expressed IDO mRNA-IFN- 纬. After treatment, the expression of tndo mRNA was significantly increased in HL-60 cell line (tn130.19) K562 cell line tnr 67.38, and both P0.05 and IFN- 纬 T 伪 1. The expression of IDO mRNA in HL-60 cell line was significantly down-regulated, and the expression of IDO protein in HL-60 cell line was significantly lower than that in HL-60 cell line (P 0.05), and the expression of IDO protein in HL-60 / K562 cell line was significantly lower than that in HL-60 / K562 cell line (P < 0.05). The expression of IDO protein in HL-60 / K562 cell line was significantly lower than that in HL-60 / K562 cell line in a dose-dependent manner. The expression level of IDO protein in HL-60 cell line was 2.505 6 卤0.019 6 after treatment with IFN- 纬, which was significantly higher than that in the control group (0.189 7 卤0.083 4 TX 326.79) P0.05 K562 cell line (0.988 8 卤0.023 9), which was significantly higher than that in the blank control group (0.253 6 卤0.016 3t76.12P0.05) IFN- 纬 T 伪 1, and the expression of IDO protein in HL-60 cell line was significantly higher than that in control group (0.988 8 卤0.023 9). After co-treatment with IFN- 纬 T 伪 1, the expression levels of ido protein in each HL-60 cell line were 2.455 卤0.066 1a 1.526 卤0.017 31.134 1 卤0.060 1U 0.8060 卤0.029 7 and 0.070 0 卤0.003 6, respectively, in a concentration-dependent manner, and the expression of Ido protein in K562 cell line was observed in a dose-dependent manner after co-treatment of IFN- 纬 T 伪 1 and IFN- 纬 T 伪 1 in K562 cell line. In a dose-dependent manner, HL-60/K562 cells induced by FN 59.50r-0.76N P0.05.IFN- 纬 could inhibit the proliferation of PHA stimulated T cells (P0.055.05), and in a concentration-dependent manner, the HL-60/K562 cells in the HL-60/K562 group and T 伪 1 group could inhibit the proliferation of T cells induced by PHA, and in a concentration-dependent manner, the proliferation of HL-60/K562 cells in the HL-60 cell line r0.981-MT and T 伪 1 group was inhibited by P0. 055. 0. 0. 7. 843 卤0.014 9, 0. 784 3 卤0.014 9, 0. 714 5 卤0.009 5. 6 卤0.007 6 卤0.007 6 6 and 0.492 2 卤0.010 9, respectively. The inhibitory rate of T cell proliferation was significantly lower than that of control group. The difference was statistically significant (P 0.05). Conclusion IDO may play an important role in the induction of immune tolerance by AML. T 伪 1 can down-regulate the expression of IDO in AML cells and improve the immunosuppressive effect of AML cells on T cells.
【作者单位】：四川省医学科学院·四川省人民医院儿科;四川省肿瘤医院肿瘤研究所;
【基金】：四川省卫生厅科研基金(090424)
【分类号】：R733.71

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