A20对急性B淋巴细胞白血病患者T淋巴细胞增殖、分化和杀伤功能的调控作用  

Regulatory effects of A20 on the proliferation,differentiation,and cytotoxicity of T lymphocytes in patients with B-cell acute lymphoblastic leukemia

作　　者：王娜 闫艳[1] 王文焕[1] WANG Na;YAN Yan;WANG Wenhuan(Department of Hematology,Bayannur Hospital,Bayannur 015000,China)

机构地区：[1]巴彦淖尔市医院血液内科,051000

出　　处：《免疫学杂志》2022年第11期964-973,共10页Immunological Journal

基　　金：巴彦淖尔市科技创新驱动联合项目(SFK2020LH01)。

摘　　要：目的观察急性B淋巴细胞白血病(B-ALL)患者A20水平,分析抑制A20表达对B-ALL患者T淋巴细胞活性的调控作用。方法收集2019年7月至2021年6月在我院就诊的B-ALL患者37例和健康对照者20例。分离外周血单个核细胞,分选CD4~+T细胞和CD8~+T细胞,流式细胞术和实时定量PCR检测A20表达。使用A20小干扰RNA(siRNA)转染B-ALL患者CD4~+T细胞和CD8~+T细胞,实时定量PCR检测CD4~+T细胞中转录因子表达,酶联斑点吸附试验检测CD8~+T细胞分泌穿孔素和颗粒酶B水平,酶联免疫吸附试验检测培养上清中细胞因子水平。组间比较使用t检验或单因素方差分析。结果B-ALL组CD4~+T细胞和CD8~+T细胞中A20~+细胞百分比、A20平均荧光强度、A20 mRNA表达均高于对照组(P<0.05);B-ALL患者CD4~+T细胞中Th1转录因子T-bet mRNA和CD8~+T细胞中穿孔素、颗粒酶BmRNA均低于对照组(P<0.05),与A20 mRNA表达呈显著正相关(P<0.05)。A20 siRNA转染不影响细胞增殖,但可增加核因子-κB p65磷酸化。A20 siRNA转染B-ALL患者CD4~+T细胞后,T-bet mRNA和分泌干扰素-γ(IFN-γ)、维甲酸相关孤独核受体γt和分泌白细胞介素-17A(IL-17A)、芳香烃受体和分泌IL-22水平较对照siRNA转染、未转染细胞均升高(P<0.05)。A20 siRNA转染B-ALL患者CD8~+T细胞后,穿孔素、颗粒酶B、IFN-γ、肿瘤坏死因子-α分泌水平较对照siRNA转染、未转染细胞均升高(P<0.05),但免疫检查点受体(PD-1、CTLA-4、TIM-3)mRNA在CD8~+T细胞中的表达水平在A20 siRNA转染、对照siRNA转染和未转染细胞中的差异无统计学意义(P>0.05)。结论B-ALL患者T细胞中A20水平升高,可能抑制T细胞活性,诱导B-ALL患者T细胞功能不全。The present study was designed to investigate A20 expression in B-cell acute lymphoblastic leukemia(B-ALL)patients,and to assess the modulatory activity of A20 to T lymphocytes.This trial recruited 37 BALL patients hospitalized in our department from July 2019 to June 2021,and 20 healthy individuals were included as controls.Peripheral blood mononuclear cells(PBMCs)were isolated,while CD4+and CD8+T cells were purified.A20 expression was measured by flow cytometry and real-time PCR.After CD4+and CD8+T cells were transfected with A20 small interfering RNA(siRNA),transcription factors in CD4+T cells were semi-quantified by real-time PCR,while perforin and granzyme B expression in CD8+T cells were assessed by enzyme linked immunospot assay.Cytokine levels in the cultured supernatants were measured by enzyme linked immunosorbent assay.Data showed that the percentage of A20+cells,A20 mean fluorescence intensity,and A20 mRNA in CD4+T cells and CD8+T cells were increased in B-ALL group,as compared with control group(P<0.05).Th1 transcription factor T-bet mRNA in CD4+T cells,perforin and granzyme B mRNA in CD8+T cells were down-regulated in B-ALL group compared with control group(P<0.05),and were positively correlated with A20 mRNA expression(P<0.05).A20 siRNA transfection did not affect cellular proliferation,but enhanced the phosphorylation of nuclear factor-κB p65.T-bet mRNA and secreting interferon-γ(IFN-γ),retinoic acid related orphan receptorγt mRNA and secreting interleukin-17A(IL-17A),as well as aryl hydrocarbon receptor mRNA and secreting IL-22 were increased in A20 siRNA transfected CD4+T cells from B-ALL patients when compared with control siRNA transfection or untransfection cells(P<0.05).The secretion of perforin,granzyme B,IFN-γ,and tumor necrosis factor-αwere elevated in A20 siRNA transfected CD8+T cells from B-ALL patients when compared with control siRNA transfection or untransfection cells(P<0.05).However,there were no significant differences in the mRNA expressions of immune checkpoint receptors(i

关 键 词：肿瘤坏死因子诱导蛋白3 急性B淋巴细胞白血病 T淋巴细胞 免疫调控 

分 类 号：R556.5[医药卫生—血液循环系统疾病]