Treg细胞延长经不同CD47基因型供者脾细胞预处理小鼠皮肤和心脏移植物存活时间的作用  

Treg cells prolong skin and heart grafts survival after different CD47 genotype donor specific splenocytes pretreatment

作　　者：王远涛[1] 胡钰[2] 高宝山[1] 王钢[1] 王伟刚[1] 傅耀文[1] 周洪澜[1] 

机构地区：[1]吉林大学第一医院泌尿外二科,长春130021 [2]吉林大学中日联谊医院病理科

出　　处：《中华器官移植杂志》2017年第6期372-377,共6页Chinese Journal of Organ Transplantation

基　　金：基金项目：国家自然科学基金（81671574,81501381,81470975）;吉林省卫生厅项目（20142042,2014Q021）;吉林省科技厅项目（20130413029GH）

摘　　要：目的 探讨调节性T淋巴细胞（Treg细胞）延长经不同CD47基因型供者脾细胞预处理小鼠皮肤和心脏移植物存活时间的作用.方法 以C57BL/6小鼠为受者,MHC-Ι错配的bm1小鼠为供者,建立MHC-Ι错配的小鼠皮肤和心脏联合移植模型.CD47^-/-DST组受鼠移植前7 d接受CD47^-/-bm1小鼠的脾细胞输注;CD47^＋/＋DST组受鼠移植前7 d接受CD47^＋/＋bm1小鼠的脾细胞输注;移植对照组受鼠仅行皮肤和心脏联合移植,不做其他预处理.术后通过观察移植皮片及异位心脏移植物的腹部触诊判断移植物存活情况,绘制移植物存活曲线;免疫组化染色检测心脏移植物内Treg细胞浸润情况,流式细胞技术检测脾脏及淋巴结中Treg细胞数量及比例的变化,混合淋巴细胞反应（MLR）检测供者特异性T淋巴细胞增殖及Treg细胞对于供者特异性T淋巴细胞增殖的抑制能力.结果 与移植对照组比较,CD47^＋/＋DST组皮肤移植物平均存活时间（MST）为20 d,比移植对照组的17.5 d略延长,差异无统计学意义（P〉0.05）;而CD47^＋/＋DST组皮肤移植物的存活时间显著延长至46.5 d（P〈0.01）.CD47^-/-DST组心脏移植物的MST为17 d,与移植对照组的15.5 d相比较,差异无统计学意义（P〉0.05）;而CD47^＋/＋DST组心脏移植物的存活时间显著延长至42.5 d（P〈0.01）.移植后18 d,CD47^-/-DST组心脏移植物内可见大量Treg细胞浸润,而同时间点的CD47^＋/＋DST组心脏移植物内未见明显Treg细胞浸润.移植后18 d,与CD47^＋/＋DST组比较,CD47^-/-DST组脾脏中Treg细胞数量显著增加（P〈0.01）,但Treg细胞的比例却明显低于CD47^＋/＋DST组（P〈0.05）;CD47^＋/＋DST组和CD47^-/-DST组淋巴结中Treg细胞数量均有增加,组间比较差异无统计学意义（P〉0.05）,但CD47^＋/＋DST组淋巴结中Treg细胞的比例明显高于CD47^-/-DST组（P〈0.01）.术后7 d的MLR显示,CD47^＋/＋DST组供者T淋巴细胞增殖明显明显受到抑制,与CDObjective To explore the role of regulatory T cells （Tregs） in skin and heart grafts survival prolongation after different CD47 genotype donor specific splenocytes pretreatment.Methods Mouse skin plus hearts transplantation model was set up by using C57BL/6 as recipients and MHC class I-mismatched bm1 as donors.In CD47^-/-DST group,recipients received CD47^-/-bm1 splenocytes transfusion at 7th d before transplantation.In CD47^＋/＋DST group,recipients received CD47^＋/＋bm1 splenocytes transfusion at 7th day before transplantation.In control group,recipients only received bm1 skin and heart grafts.The populations of Tregs were analyzed by FACS and immunohistochemistry,respectively.The inhibitory effect of Tregs and anti-donor T cell responses was assessed by MLR.Results Result As compared with control group,the survival time of skin grafts in CD47^-/-DST group was slightly longer than in non-DST group （20 days vs.17.5 days,P〉0.05）,but skin grafts had long-term survival in CD47^＋/＋DST group （46.5 days,P〈0.005）.The survival time of heart grafts in CD47^-/-DST group was similar to that in the control group,and slightly longer than non-DST group （17 days vs.15.5 days,P〉0.05）,but heart grafts had long-term survival in CD47^＋/＋DST group （42.5 days vs.17 days,P〈0.01）.At 18th day after transplantation,immunohistochemistry showed that more Tregs infiltration could be observed in CD47^-/-DST group,but not in CD47^＋/＋DST group.The number of Foxp3＋Tregs from the mice receiving CD47^-/-DST increased markedly at 18th day post-transplantation compared to thot from the mice receiving CD47^＋/＋DST （P〈0.01）,but the ratio of Tregs was decreased significantly （P〈0.05）.The number of Foxp3＋Tregs in both CD47^-/-and CD47^＋/＋ DST groups was increased,but there was no significant difference between two groups （P〉0.05）.As compared with CD47^-/-DST group,the ratio of Tregs in lymph node cells in CD47^＋/＋DST group increased significantly （P〈0.01）.Compared to

关 键 词：T淋巴细胞 调节 皮肤移植 心脏移植 联合移植 小鼠 移植物存活 

分 类 号：R617[医药卫生—外科学]