通过小鼠皮肤移植后心脏移植模型观察记忆性T淋巴细胞的变化和作用  

The role of memory T cells in acute rejection of heart transplantation following skin transplantation

作　　者：梁华[1] 廖崇先[1] 林智[1] 舒涛[1] 杨谦[1] 单忠贵[2] 

机构地区：[1]厦门大学附属中山医院厦门市心脏中心,福建厦门361000 [2]厦门大学附属第一医院

出　　处：《中华器官移植杂志》2012年第3期170-173,共4页Chinese Journal of Organ Transplantation

基　　金：厦门市科技计划项目（3502220094011）;2008年福建省科技计划重点项目（闽科计[2008]59）

摘　　要：目的建立小鼠皮肤移植后心脏移植模型，观察记忆性T淋巴细胞的变化和作用，及其与皮肤移植后同种反应性记忆性T淋巴细胞过继转移模型的差异和机理。方法应用显微外科技术及血管套管法，建立小鼠皮肤移植后心脏移植模型（实验组），并以心脏移植前输注或未输注记忆性T淋巴细胞的2个组作为对照（移植对照组和输注对照组），两对照组均未进行皮肤移植。心脏移植后每天触摸各组供心的搏动情况，观察各组移植心的存活情况及组织病理学改变，并对排斥反应进行分级；采用流式细胞仪检测各组受鼠体内CD3＋CD44highCD62L-记忆性T淋巴细胞表型的分布，应用体外混合淋巴细胞反应体系综合评价各组记忆性T淋巴细胞体外增殖程度。结果皮肤移植前受鼠体内CD4＋记忆性T淋巴细胞的比例为2．7％，移植后升至72．0％（P〈0．05）。移植对照组、输注对照组及实验组移植心脏的平均存活时间分别为7．33、4．83和3．5d，排斥反应评分分别为1B～2级、3A级和4级，体内CD3＋CD44highT淋巴细胞的比例分别为29％、55％和75％，CD3＋CD62L—T淋巴细胞群落中CD44荧光强度分别为215、274和380，以及MLR强度（吸光度值）分别为0．29、0．45和1．0，与两对照组相比，实验组移植心存活时间明显缩短、记忆性T淋巴细胞比例明显升高、MLR强度均明显增强及排斥反应程度更严重（P〈0．05或P〈0．01）。结论同种小鼠皮肤移植后心脏移植模型是一个更为贴近临床实际的动物模型，其同种抗原反应能力明显强于过继输注同种记忆性T淋巴细胞的小鼠心脏移植模型，有利于免疫记忆的相关研究。Objective The presence of alloreactive memory T cells in recipient is a critical handicap to achieve transplantation tolerance. To make a mouse model which mimics the present transplant patient is important for research at this subject. Thus, we developed a novel re-transplant model and compared the alloresponse in this model with that in the conventional memory T cells-transfer model （transfer control）. Methods The re-transplant model was established via microsurgery and vessel cannula techniques, and the experiment was composed of three groups： the retransplant group, memory T cell-transfer group （transfer control） and the conventional blank group （blank control）. The research indexes included survival time of donor heart, rejection score of allograft, and detection of proliferation and differentiation of the alloreactive memory/effector T cells by by flow cytometry （FCM） and in vitro mixed lymphocyte reaction （MLR）. Results The median survival time of allograft in re-transplant recipients was significantly shortened compared to that of transfer control, but there was no significant difference in rejection score of graft between them （the score in re- transplant group was the most intense of the three groups）. Moreover, proliferation and differentiation of the alloreactive effector T cells were more intensive in re- transplant recipients than in the transfer control, which was confirmed by in vitro MLR and by FCM of the splenocytes for detecting CD44highCD62L-memory/effeetor phenotype cells. Conclusion The recall alloresponse in retransplantation is more intensive than that in memory-transfer setting and this re-transplant model is more close to the clinic situation than the memory-transfer model in rodents.

关 键 词：小鼠 皮肤移植 心脏移植 记忆性T淋巴细胞 

分 类 号：R654.2[医药卫生—外科学]