应用iDC从G-CSF动员的外周血中诱导产生CD4^＋CD25^＋调节性T细胞  

Characteristics of induced CD4^+CD25^+ regulatory T lymphocytes in G-CSF mobilized peripheral blood by iCD

作　　者：苏虹[1] 王忱诚1 王保龙[2] 彭美玲[1] 

机构地区：[1]安徽医科大学公共卫生学院流行病与卫生统计学系,合肥230032 [2]安徽医科大学附属省立医院检验科,合肥230001

出　　处：《安徽医科大学学报》2012年第5期500-504,共5页Acta Universitatis Medicinalis Anhui

基　　金：安徽省自然科学基金面上项目(编号:11040606M172);安徽医科大学博士科研资助基金(编号:XJ200819)

摘　　要：目的探讨在粒细胞集落刺激因子(G-CSF)动员的外周血单个核细胞(PBMC)中诱导产生CD4+CD25+调节性T细胞(Treg)的可行性及其表型和功能。方法收集G-CSF动员和未动员的BALB/c♀鼠的PBMC,采用磁活性标记的细胞分选法(MACS)分选BALB/c♀鼠的CD4+CD25-T细胞和CD4+CD25+T细胞及C57BL/6♂鼠的CD11c+未成熟树突细胞(iDC),建立细胞培养体系,经iDC体外诱导G-CSF动员和未动员的CD4+CD25-T细胞转换为CD4+CD25+Treg细胞(iTreg)。分别应用流式细胞术、混合淋巴细胞反应技术检测诱导后细胞的CD25、Foxp3的表达水平以及抑制功能,比较G-CSF动员与非动员组间iTreg的表型和抑制功能的差异。结果 iDC诱导G-CSF未动员和动员的组间CD25+分子表达率分别为(76.8±4.1)%和(90.4±5.3)%,差异有统计学意义(P<0.01);两组诱导产生的CD25+T细胞的Foxp3表达水平分别为(64.1±2.7)%和(59.5±3.2)%,差异有统计学意义(P<0.05)。iDC诱导G-CSF未动员和动员的外周血产生的iCD4+CD25+Treg均表现出免疫抑制功能,其中动员后诱导生成的iTreg的免疫抑制效应明显增强。结论应用iDC从G-CSF动员的外周血中诱导产生的iCD4+CD25+Treg具有更强的体外抑制效应,为自身免疫性疾病的治疗提供新的思路和手段。Objective To explore the feasibility of inducing CD4^＋CD25^＋regulatory T lymphocytes （Tregs） from mononuclear cells of granulocyte-colony stimulating factor （G-CSF） mobilized peripheral blood, and to analyze the phenotype and function of these cells. Methods G-CSF mobilized and un-mobilized peripheral blood mononuclear cells were collected from BALB/c murine. CD4^＋CD25 - T cells and CD4^＋CD25^＋T cells of BALB/c murine, and CD11c^＋ cells （immature dendritic cell, iDC） of C57BL/6 murine were separated with magnetic activated cell sorting （MACS） respectively. Different cell culture systems were set up to induce CD4^＋CD25^＋Tregs （iTregs） by iDC from mononuclear cells of G-CSF mobilized and un-mobilized peripheral blood. Then flow cytometry （FCM） and mixed lymphocyte reaction （MLR） were used respectively to test the expression of CD25^＋, Foxp 3 and suppres-sive function of these iTregs. The differences of the rate and the suppressive effect were compared among different groups. Results After treated with iDC, there was significant difference in the expression Of CD25^＋between different groups with CD4^＋CD25^- T cells isolated from G-CSF mobilized and un-mobilized groups, which were（90.4 ± 5.3 ） % and （76. 8 ±4. 1 ） %, respectively （ P 〈 0.01 ）. The expressions of Foxp3 of the above groups were （ 64. 1 ± 2.7 ）% and （59.5 ± 3.2）% respectively, the difference was also significant （P 〈 0. 05 ）. ＇The iTregs came from G- CSF mobilized peripheral blood induced by iDC had more suppressive effect compared with the unmobilized group in vitro, the difference was significant （P 〈 0. 01 ）. Conclusion The induced CD4^＋CD25^＋Tregs in G-CSF mobi- lized peripheral blood by iDC have more suppressive effect in vitro. It indicates that G-CSF mobilized peripheral blood can serve as an important source of CD4^＋CD25^＋Tregs, which has potential to be used as an immunotherapy agent for autoimmune diseases.

关 键 词：粒细胞集落刺激因子 未成熟树突细胞 CD4+CD25+调节性T细胞 叉状头/翅膀状螺旋转录因子 

分 类 号：R446.6[医药卫生—诊断学]