肥大细胞体外诱生CD4^＋CD25^＋Foxp3^＋调节性T细胞的作用  

Mast cells induce CD4^+CD25^+Foxp3^+ regulatory T cells in vitro

作　　者：张维娜[1] 吴轲[1] 周鸿敏[1] 何文涛[1] 高英[1] 汪理[1] 林星光[1] 方泽民[1] 蔡兰军[1] 周巧丹[1] 雒真龙[1] 陈忠华 

机构地区：[1]华中科技大学同济医学院附属同济医院器官移植研究所 教育部/卫生部重点实验室,武汉430030

出　　处：《现代免疫学》2009年第6期453-457,共5页Current Immunology

基　　金：国家自然科学基金资助项目(30772038、36071989);国家重点基础研究发展规划(973计划)(2009CB522407)

摘　　要：探索小鼠骨髓源性肥大细胞能否在体外诱生CD4+CD25+Foxp3+调节性T细胞。从小鼠股骨获取骨髓细胞,加入完全RPMI 1640培养基(含IL-3和SCF各10 ng/ml)诱导4周。用甲苯胺蓝染色法观察肥大细胞异染颗粒;流式检测CD117和FcεRIα双阳性细胞比例。将肥大细胞与同基因来源的T细胞按不同比例(1∶2、1∶1、2∶1)置于48孔培养板中培养,作为三个实验组;未加肥大细胞的单独T细胞组作对照组。四个组均加入CD3抗体和CD28抗体各2μg/ml,IL-2 1 000U/ml,5 d后流式检测Foxp3表达情况。RT-PCR,免疫组化法检测肥大细胞中TGF-β1的表达。与对照组相比,实验组Foxp3表达均升高(对照组:3.37%±0.40%;实验组为:8.23%±0.80%、10.87%±1.25%、13.63%±0.55%)。RT-PCR和免疫组化均检出TGF-β1表达。肥大细胞能在体外诱导T细胞转化为CD4+CD25+Foxp3+调节性T细胞,可能与肥大细胞表达TGF-β1有关。To detect whether mouse mast cells can induce CD4^＋ CD25^＋ Foxp3^＋ regulatory T ceils （Treg） in vitro. Bone marrow cells obtained from C57BL/6 （H-2b） mice were cultured with IL 3 （10 ng/ml） and SCF （10 ng/ml） for 4 weeks. The purity of bone marrow mast cells （BMMCs） was tested by flow cytometry. The expression of TGF-β1 in BMMCs was tested by RT-PCR and immunohistochemistry. Then the BMMCs were co-cultured with T cells of C57BL/6 mice at 1 ： 2, 1 ： 1 and 2 ： 1 ratios in the presence of anti-CD3 antibody（2μg/ml）, anti-CD28 antibody（2 μg/ml） and IL 2 （1 000 U/ml）. The percentages of CD4^＋ CD25^＋ Foxp3^＋ T cells in the co-cultured system were compared among different groups by flow cytometry on day 5. It was found that the percentages of CD4^＋ CD25 ^＋ Foxp3^＋ cells were significantly higher in the ratio of BMMC/T 2 ： 1 group（13.63% ±0.55%）, the 1 ： 1 group（10.87% ± 1.25%）and 1 ： 2 group（8.23% ± 0.80%） than in the control group（3.37%± 0.40 %）. The expression of TGF-β1 was determined in the mouse BMMCs by RT-PCR and immunohistochemistry. Our findings indicate that CD4^＋ T cells can be induced to become CD4^＋ CD25^＋ Foxp3^＋ Treg by BMMCs in vitro and the secretion of TGF-β1 by BMMCs might be involved in this process.

关 键 词：小鼠 肥大细胞 CD4^+CD25^+FOXP3^+调节性T细胞 TGF-Β1 

分 类 号：R392.4[医药卫生—免疫学]