Renca条件培养基转化的调节性T细胞对小鼠同种免疫的影响  

Effect of Converted Regulatory T Cells with Renca Conditioned Media on Suppressive Profile of Allo-immunity in Mice

作　　者：滕立臣[1] 刘龙山[1] 苏亚娟[2] 袁小鹏[1] 李军[1] 付倩[1] 陈思阳[3] 王长希[1] 

机构地区：[1]中山大学附属第一医院器官移植科,广东广州510080 [2]南方医科大学附属南方医院检验系,广东广州510015 [3]中山大学附属第一医院麻醉科,广东广州510080

出　　处：《中山大学学报（医学科学版）》2010年第2期242-248,共7页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：中山大学海外岭南基金(80000-9390001)

摘　　要：【目的】探讨小鼠肾癌细胞株Renca来源的条件培养基能否诱导CD4+CD25-T细胞转化为具有免疫抑制效能的CD4+CD25+Foxp3+T细胞。【方法】培养Renca提取上清获得条件培养基,将条件培养基与普通培养液按不同比例混合培养小鼠CD4+CD25-T细胞7d,以流式细胞仪检测CD4+CD25+T细胞和Foxp3+细胞的比例;再次分选CD4+CD25+T细胞检测其抑制细胞增殖的能力,体内过继性输入观察对移植物存活时间、局部排斥反应和迟发性超敏反应的影响。【结果】与对照组相比,混合培养基显著增加培养体系中CD4+CD25+Foxp3+T细胞的比例(P<0.05),在一定范围内(≤75%)呈比例依赖关系;随转化性CD4+CD25+T细胞比例增加效应T细胞增殖能力减弱;体内输注显著延长移植物的存活时间(P<0.05),达(29.6±1.4)d,组织学观察显示能明显抑制局部排斥反应。【结论】Renca条件培养基能诱导CD4+CD25-细胞转化为CD4+CD25+Foxp3+T细胞,被转化细胞在体内和体外均能发挥免疫抑制作用。【Objective】 This study was designed to investigate capability of the conditioned media that originated from Renca cells to convert CD4^＋ CD25^- T cells into CD4^＋ CD25^＋ T cells, which can exert immunosuppressive effect on effector T cells in vitro and in vivo. 【Methods】 The common media were mixed with the conditioned media at different ratios, and fresh enriched CD4^＋ CD25^- T cells with MACS were cultured in mixed media for 7 days. At end-point of culture, the cells were collected and detected phenotypes in flow cytometer. Moreover, we detected immunosuppressive effect of converted CD4^＋ CD25^＋ T cells on effector T cells proliferation in one-way mixed lymphocytes reaction by using CCK-8, and we observed survival time and histology of grafts. The delayed type hypersensitivity was determined 14 days after transplantation. 【Results】 The mixed media could increase ratio of CD4^＋ CD25^＋ Foxp3^＋ T cells in conditioned media ratio-dependent （P 0.05）, compared with control groups, when the mixed media contained no more than 75% of conditioned media. The converted CD4^＋ CD25^＋ T cells significantly suppress proliferation of effector T cells in vitro, and prolong survival time of grafts, which were （29.6 ± 1.4） d in converted CD4^＋ CD25^＋ T cells treated groups （P 0.05）, compared with that in untreated groups （9.8 ± 0.6 d） or PBS treated groups （10.9 ± 0.6 d）. Moreover, delayed-type hypersensitivity reaction were conducted at day 14 after transplantation in the recipients, and the results showed that less pad swelling in the group treated with converted CD4^＋ CD25^＋ T cells than other control groups was found, according to measurement of pad swelling. In addition, progressed to complete necrosis of grafts were exhibited in the mice treated with PBS and untreated mice, whereas better healing of grafts and less lymphocytes infiltration were displayed in the mice treated with converted CD4^＋ CD25^＋ T cells, which were similar to the mice treated with nat

关 键 词：条件培养基 转化 调节性T细胞 免疫抑制 

分 类 号：R617[医药卫生—外科学]