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目的:研究急性髓系白血病(AML)中辅助性T细胞9(T helper cell,Th9)的分布及其与疾病临床特征的关系,同时研究Th9中不同转录因子的激活情况。方法:收集102名AML患者及83名健康人外周血样本,应用CD3磁珠法分离健康人及AML患者外周血T细胞,应用实时定量PCR检测IL-9的mRNA表达,应用流式细胞术检测CD4~+IL-9~+(Th9)细胞在疾病各阶段的比率,以及与IL-9共表达的转录因子激活情况。结果:AML-M2和M3型患者外周血T细胞中IL-9的mRNA表达显著高于健康对照者(P<0.01),同时CD4~+IL-9~+细胞比率也明显高于健康对照者(P<0.01)。对M2和M3中Th9细胞分布动态监测结果表明,初诊M2和M3及复发M2组Th9细胞比率及IL-9 mRNA表达显著高于缓解状态下M2和M3患者(P<0.01)。对调控IL-9表达的转录因子共表达分析表明,初诊及复发M2及M3患者的外周血中p-SMAD3~+的Th9细胞显著多于缓解时M2及M3患者(P<0.01);而与之相反,缓解期M2及M3患者外周血中IRF-1~+的CD4~+IL-9~+细胞显著多于初诊M2及M3患者(P<0.01)。结论:辅助性T细胞Th9细胞在AML-M2和M3中分布显著增高,并且与疾病状态相关;对于Th9细胞的功能预测需要与其转录因子结合进行,后者对AML患者体内肿瘤微环境具有重要意义。
Objective: To investigate the distribution of T helper cell (Th9) in acute myeloid leukemia (AML) and its relationship with the clinical features of the disease, and to study the activation of different transcription factors in Th9. Methods: Peripheral blood samples of 102 AML patients and 83 healthy volunteers were collected. CD3 magnetic beads were used to separate peripheral blood T cells from healthy people and AML patients. Real-time quantitative PCR was used to detect the mRNA expression of IL-9. Flow cytometry The ratio of CD4 ~ + IL-9 ~ + (Th9) cells at various stages of the disease and the activation of transcription factors co-expressed with IL-9. Results: The mRNA expression of IL-9 in peripheral blood T cells of AML-M2 and M3 patients was significantly higher than that of healthy controls (P <0.01), and the ratio of CD4 ~ + IL-9 ~ + cells was also significantly higher than that of healthy controls (P <0.01). The dynamic monitoring results of Th9 cell distribution in M2 and M3 showed that the ratio of Th9 cells and the expression of IL-9 mRNA in newly diagnosed M2 and M3 and relapsed M2 patients were significantly higher than those in M2 and M3 patients in remission (P <0.01). Co-expression analysis of transcription factors that regulate IL-9 expression showed that peripheral blood p-SMAD3 + + Th9 cells in newly diagnosed and recurrent M2 and M3 patients were significantly more than those in M2 and M3 patients at remission (P <0.01) On the contrary, the number of CD4 ~ + IL-9 ~ + cells in peripheral blood of patients with M2 and M3 remission was significantly higher than that of newly diagnosed patients with M2 and M3 (P <0.01). CONCLUSIONS: Th9 cells of helper T cells are significantly increased in AML-M2 and M3, and are associated with disease states; functional predictions of Th9 cells need to be performed in conjunction with their transcription factors, which are important for the in vivo tumor microenvironment of AML patients .