【摘要】：目的檢測CD4~+CD25~+Fox P3~+調(diào)節(jié)性T細胞(Treg)在兒童急性B淋巴細胞白血病(pre-B ALL)外周血樣本中的比例,探討Helios基因在pre-B ALL外周血Treg細胞中的功能。方法 ALL組選取10例初發(fā)pre-B ALL外周血標本,對照組選取6例行骨關(guān)節(jié)病手術(shù)并排除腫瘤、血液系統(tǒng)、免疫系統(tǒng)疾病的患兒外周血標本;采用流式細胞術(shù)檢測Treg細胞在pre-B ALL中CD4~+細胞的比例,通過定量PCR技術(shù)檢測Helios、Fox P3及相關(guān)轉(zhuǎn)錄抑制因子在ALL中CD4~+CD25~+Treg細胞中的表達;通過慢病毒轉(zhuǎn)染和siRNA技術(shù)在Treg細胞中調(diào)節(jié)Helios表達,檢測Tregs免疫抑制功能的變化。結(jié)果 ALL組中CD4~+CD25~+Fox P3~+細胞占CD4~+T細胞比例[(6.16±0.79)%]顯著高于對照組[(2.62±0.34)%](P=0.005);Helios、Fox P3、TGF-β1 mRNA在ALL患者Treg細胞中上調(diào)表達;Helios基因的異常表達與Treg細胞的免疫抑制功能呈正相關(guān)。結(jié)論兒童pre-BALL發(fā)病過程中,免疫功能異常與Treg細胞數(shù)量異常相關(guān)。其中Helios基因的異常表達可通過影響Treg細胞的免疫抑制功能,間接影響白血病患兒的免疫調(diào)控。Helios作為靶基因可能對以Treg細胞為主的免疫治療具有治療意義。
[Abstract]:Objective to detect the proportion of CD4 ~ CD25 ~ Fox P3- ~ regulatory T cell (Treg) in peripheral blood samples of children with acute B lymphocytic leukemia (pre-B ALL) and to explore the function of Helios gene in pre-B ALL peripheral blood Treg cells. Methods the peripheral blood samples of 10 patients with primary pre-B ALL in the ALL group and 6 children with osteoarthropathy were selected in the control group, and the peripheral blood samples of the children with tumor, blood system and immune system diseases were excluded. The percentage of Treg cells in pre-B ALL was detected by flow cytometry, and the expression of Helios Fox P3 and related transcription suppressor factor in ALL was detected by quantitative PCR technique. The expression of Helios in Treg cells was regulated by lentivirus transfection and siRNA technique, and the immunosuppressive function of Tregs was detected. Results the percentage of CD4 ~ CD25 ~ Fox P3~ cells in ALL group [(6.16 鹵0.79)%] was significantly higher than that in control group [(2.62 鹵0.34)%] (P0. 005). The up-regulated expression of Helios gene in Treg cells of ALL patients was positively correlated with the immunosuppressive function of Treg cells. Conclusion the abnormal immune function is correlated with the abnormal number of Treg cells in children with pre-BALL. The abnormal expression of Helios gene may affect the immunosuppressive function of Treg cells, and indirectly affect the immune regulation of leukemia children. Helios as a target gene may have therapeutic significance for immunotherapy with Treg cells.
【作者單位】： 山東大學深圳研究院;山東大學齊魯醫(yī)院兒科;
【基金】：深圳市科技研發(fā)基金知識創(chuàng)新計劃(JCYJ2014 0418115449178) 山東省科技發(fā)展計劃(2014GSF118131) 山東大學基本科研業(yè)務費(2014QLKY02&2014QY003-11)
【分類號】：R733.71
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本文編號：2193396