TGF—β1诱导的调节性T细胞体内输注延长小鼠皮肤移植物存活  

Transfer of regulatory T cells induced by TGF-β1 prolonged the skin-graft survival in mice

作　　者：袁劲[1] 吴轲[2] 何文涛[2] 汪峰[2] 陈忠华 

机构地区：[1]华中科技大学同济医学院同济医院肾病内科,武汉430030 [2]器官移植研究所器官移植教育部/卫生部重点实验室

出　　处：《中华微生物学和免疫学杂志》2008年第8期680-684,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金资助项目（30671989）

摘　　要：目的利用TGF-β1体外诱导naiveT细胞分化为调节性T细胞（Treg），通过体内输注延长小鼠皮肤移植物存活时间，并研究其相关机制。方法根据诱导条件不同分为3组：对照组（加入IL-2培养的C57BL／6小鼠T细胞）、MLR组（即混合淋巴细胞反应组，经同种抗原刺激活化的C57BL／6小鼠T细胞）和TGF-β组（经同种抗原刺激活化的C57BL／6小鼠T细胞，同时加入5．0ng／ml TGF-β1诱导）。利用FACS检测CD4^＋CD25^＋T细胞比例，并用RT—PCR检测Foxp3的表达水平。建立小鼠皮肤移植模型，并于第0、1、2和3天输注上述细胞，观察皮肤移植物存活时间。术后第9天，取部分受鼠行移植物病理检测，用FACS检测脾脏中TH1、TH2和CD4^＋CD25^＋Treg比例，并用AlamarBlue法检测淋巴细胞增殖能力。结果TGF-β组中CD4^＋CD25^＋T细胞比例高于对照组和MLR组（P〈0．05），且其高表达Foxp3。将培养的细胞输注给受鼠后发现，输注MLR组细胞的受鼠其移植物平均存活时间（mean survival time，MST）为（9．4±1．3）d，低于对照组（P〈0．05）；而输注TGF-β组细胞小鼠MST为（22．8±1．9）d，较对照组和MLR组明显延长（P〈0．05）。病理检测亦显示TGF-β组受鼠移植物结构完整，无明显淋巴细胞浸润。FACS结果显示TGF-β组小鼠体内TH1细胞（CD4^＋TIM-3^＋）比例低于对照组和MLR组（P〈0．05），而TH2细胞（CD4^＋TIM-1^＋）比例则与MLR组类似（P〉0．05），但低于对照组（P〈0．05）；而且TGF-β组中CD4^＋CD25^＋Treg比例明显高于对照组和MLR组（P〈0．05）。用AlamarBlue法检测受鼠外周淋巴细胞增殖活性显示，TGF-β组受鼠淋巴细胞增殖能力被明显抑制，低于对照组（P〈0．05）。结论TGF-β，可诱导T细胞分化为具有抑制能力的Treg，将诱导后的细胞进行过继输注可使小鼠体内CD4^＋CD25^＋T。比例升高，同时抑制TH1和TH2细胞分化扩增，并�Objective To study the probability of transferring the regulatory T cells induced by TGF-β1 to prolong the allograft survival and the mechanisms involved. Methods According to the different culture conditions, three experimental groups were established： control group （T cells from C57BL/6 mice cultured with IL-2）, MLR group（ T cells from C57BL/6 mice activated by alloantigen） and TGF-β group （T cells from C57BL/6 mice activated by alloantigen and cultured with 5.0 ng/ml TGF-β1 ）. After the culture, the ratio of CD4^＋CD25^＋ T and the Foxp3 expression were measured by FACS and RT-PCR, respectively. On 9th day, the pathologic analysis was performed and the ratios of TH1, TH2 and T reg and the proliferation of lymphocytes were measured. Results The ratio of CD4^＋ CD25^＋ T in TGF-β group was higher than that in control group and MLR group （ P 〈 0.05 ） , and Foxp3 was expressed in CD4^＋ CD25^＋ T cell from TGF-β group. After transferring of the cells, the allograft survival time in TGF-β group was prolonged and its mean survival time （MST） was （22.8 ± 1.9） d, which was longer than that in MLR group and control group （ P 〈 0.05 ）, but MST of MLR group was （9.4 ± 1.3 ） d, which was shorter than that of control group （ P 〈 0.05）. In TGF-β group, the ratio of TH1 was lower than that of control group and MLR group （P 〈0.05）, but the ratio of TH2 was similar with MLR group （ P 〉 0.05 ）, lower than that of control group （ P 〈 0.05 ）. And the ratio of CD4 ^＋ CD25^＋ T in TGF-β group was higher than that in control and MLR group obviously （ P 〈 0.05）. In addition, the proliferation of lymphocytes in TGF-β group was impaired. Conelusion TGF-β1 could induce naive T cells to differentiate into regulatory T cells. After transferring these cells in mice skin transplant model, the ratio of CD4^＋ CD25^＋ Treg was increased, and the differentiation of TH1 and TH2 and the proliferation of lymphocytes were inhibited. Finally, the allograft su

关 键 词：TGF—β1 调节性T细胞 皮肤移植 

分 类 号：R392[医药卫生—免疫学]