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目的检测CD4~+CD25~+Fox P3~+调节性T细胞(Treg)在儿童急性B淋巴细胞白血病(pre-B ALL)外周血样本中的比例,探讨Helios基因在pre-B ALL外周血Treg细胞中的功能。方法 ALL组选取10例初发pre-B ALL外周血标本,对照组选取6例行骨关节病手术并排除肿瘤、血液系统、免疫系统疾病的患儿外周血标本;采用流式细胞术检测Treg细胞在pre-B ALL中CD4~+细胞的比例,通过定量PCR技术检测Helios、Fox P3及相关转录抑制因子在ALL中CD4~+CD25~+Treg细胞中的表达;通过慢病毒转染和siRNA技术在Treg细胞中调节Helios表达,检测Tregs免疫抑制功能的变化。结果 ALL组中CD4~+CD25~+Fox P3~+细胞占CD4~+T细胞比例[(6.16±0.79)%]显著高于对照组[(2.62±0.34)%](P=0.005);Helios、Fox P3、TGF-β1 mRNA在ALL患者Treg细胞中上调表达;Helios基因的异常表达与Treg细胞的免疫抑制功能呈正相关。结论儿童pre-BALL发病过程中,免疫功能异常与Treg细胞数量异常相关。其中Helios基因的异常表达可通过影响Treg细胞的免疫抑制功能,间接影响白血病患儿的免疫调控。Helios作为靶基因可能对以Treg细胞为主的免疫治疗具有治疗意义。
Objective To detect the proportion of CD4 ~ + CD25 ~ + Fox P3 ~ + regulatory T cells (Treg) in peripheral blood of children with acute B-cell lymphocytic leukemia (pre-B ALL) and to explore the effect of Helios gene on pre-B ALL peripheral blood Treg Function in cells. Methods Peripheral blood samples of 10 pre-B ALL patients were selected from the ALL group, 6 from the control group were selected for osteoarthrosis surgery and were excluded from the tumor, hematological system and immune system diseases. Flow cytometry was used to detect Treg The percentage of CD4 ~ + cells in pre-B ALL cells was detected by quantitative PCR. The expression of Helios, Fox P3 and related transcriptional repressors in CD4 ~ + CD25 ~ + Treg cells were detected by quantitative PCR. Technology regulates Helios expression in Treg cells and detects changes in immunosuppressive function of Tregs. Results The percentage of CD4 ~ + CD25 ~ + Fox P3 ~ + CD4 ~ + T cells in ALL group was significantly higher than that in control group [(2.62 ± 0.34)%] , Fox P3 and TGF-β1 mRNA were upregulated in Treg cells of ALL patients. The abnormal expression of Helios gene was positively correlated with the immunosuppressive function of Treg cells. Conclusion In the process of pre-BALL in children, abnormal immune function is related to abnormal Treg cell number. The abnormal expression of Helios gene can indirectly affect the immune regulation of children with leukemia by affecting the immunosuppressive function of Treg cells. Helios as a target gene may have therapeutic implications for Treg cell-based immunotherapy.