胸腺上皮细胞可植入异基因骨髓移植小鼠胸腺并促进T淋巴细胞数量重建  

Establishment of a animal model of allogeneic bone marrow transplantation plus thymic epithelial cellstransplantation

作　　者：梁惠[1] 吴翠玲[1] 陈镜如 张玉明[1] 

机构地区：[1]南方医科大学南方医院小儿科,广州510515

出　　处：《中华器官移植杂志》2017年第11期687-691,共5页Chinese Journal of Organ Transplantation

基　　金：国家自然科学基金（81270632）

摘　　要：目的 通过建立异基因骨髓移植（BMT）联合胸腺上皮细胞移植（TCT）小鼠模型,探讨TCT应用于BMT的可行性及其效应.方法 以BALB/C小鼠为受鼠,C57BL/6小鼠为供鼠,移植前1d对受鼠进行全身照射预处理,BMT后取C57BL/6孕鼠胚胎期14～16 d胎胸腺上皮细胞进行TCT.为了分析合适的全身照射剂量,将接受不同全身照射剂量的受鼠分为3组：7 Gy组、6.5 Gy组和6 Gy组.为了分析胸腺上皮细胞植入情况,以接受BMT联合供鼠来源TCT的受鼠作为实验组,以单纯接受BMT的受鼠作为对照组,每组8只.采用活体成像检测分析胸腺上皮细胞植入情况,测定胸腺指数,取各组受鼠胸腺进行组织学检查及观察胸腺上皮细胞表面标记分子K8、K5免疫荧光抗体染色情况,应用流式细胞仪检测各组受鼠移植4周后外周血T淋巴细胞亚群.结果 接受6.0 Gy剂量的受鼠能够长期存活但植入失败,照射剂量6.5 Gy时受鼠的存活率降低但BMT和TCT皆植入成功,表明6.5 Gy是最小致死剂量,作为建立模型的全身放疗剂量.活体成像检测显示,移植后2周实验组受鼠前胸部皮下可检测到明显的荧光信号,对照组受鼠未检测到任何荧光.移植4周,实验组受鼠的胸腺单叶明显比对照组单叶体积大,且胸腺指数也较高.组织学观察显示,嵌合了异基因TEC的受鼠胸腺具有典型的胸腺皮质和髓质结构,且明显表达K8＋和K5＋胸腺髓质上皮细胞.移植4周后,实验组CD4＋和CD8＋T淋巴细胞比例均明显高于对照组（P＜0.05）.结论 胸腺上皮细胞可植入异基因BMT受鼠胸腺内,并促进受鼠外周免疫T淋巴细胞数量的重建.Objective To explore the establishment of animal model of allogeneic bone marrow transplantation plus thymic epithelial cells transplantation,and then examine the feasibility and effects of thymic epithelial cells transplantation applied in allogeneic bone marrow transplantation.Methods One day before transplantation the recipient BALB/C mice were given total-body irradiation,then transplanted with bone marrow cells from donor C57BL/6 mice and thymic epithelial cells from E14-16 embryonic thymus of donor C57BL/6 mice.In order to explore the appropriate irradiation dose,we set up three different dose groups：7 Gy;6.5 Gy;6 Gy.The recipient mice transplanted with BMT plus TCT served as experimental group,and those transplanted with BMT only served as control group （n =8 each）.Then in vivo imaging in mammals was done to observe the thymic epithelial cells transplantation.Thymus index was measured.The thymus in each group was collected for histological examination and immunohistochemical staining of K5 and K8.Flow cytometry was used to examine the T cells subsets in peripheral blood of recipient mice 4 weeks after thymus transplantation.Results The recipient mice with 6.0 Gy TBI had long-term survival but implantation was done unsuccessfully,and those with 6.5 Gy had lower survival rate but implantation was done successfully.6.5 Gy was the minimum lethal dose and could be used as the appropriate irradiation does in this study.In vivo imaging in mammals detecting system showed the experimental group obvious fluorescent signals could be detected in the experimental group,but no fluorescence was found in the control group.Four weeks after transplantation,the thymus was bigger and thymus index was higher in the experimental group than in the control group.And the chiemra thymus of the experimental group also had normal cortex and medulla histological structure.Four weeks after transplantion,the percentages of CD4＋ and CD8＋ T cells of the peripheral blood in experimental group were significantly higher than in con

关 键 词：小鼠 造血干细胞移植 胸腺上皮细胞 动物模型 免疫重建 

分 类 号：R457.7[医药卫生—治疗学]