供体淋巴细胞在GVHD模型小鼠组织器官中迁移和分布的动态观察  

Migration and Distribution of Allogeneic T Lymphocytes in Organs of Graft-Versus-Host Disease Mouse Model

作　　者：温宏升[1] 王健民[1] 周虹[1] 夏荣[1] 邱慧颖[1] 高磊[1] 胡晓霞[1] 

机构地区：[1]第二军医大学长海医院血液科,上海200433

出　　处：《中国实验血液学杂志》2006年第5期919-923,共5页Journal of Experimental Hematology

基　　金：国家自然科学基金资助项目(编号39870710;30172347);上海市卫生系统百名跨世纪优秀学科带头人计划(编号98BR029)

摘　　要：本研究应用小鼠GVHD模型探讨异基因T淋巴细胞在移植受体体内的迁移和分布。将C57BL/6的骨髓细胞和转基因荧光C57BL/6小鼠的脾淋巴细胞输入经8Gy全身照射的BALB/c小鼠,建立eGFP标记供体淋巴细胞的GVHD小鼠模型;应用荧光显微镜和流式细胞术观测GVHD模型中eGFP+细胞的分布;ELISA法检测GVHD靶组织中趋化因子MIP-1α水平的改变。结果表明:①输入脾细胞和骨髓细胞第8天后出现GVHD临床及病理表现;②GVHD模型中,受体鼠肝、皮肤、肠、脾、肺、舌有eGFP+细胞浸润;③GVHD小鼠肝、脾eGFP+细胞中CD4+、CD8+细胞比例均逐渐升高;④GVHD鼠脾、肝组织中MIP-1α水平升高,脾中MIP-1α水平高峰出现于移植后第3天,肝中MIP-1α水平高峰出现于移植后第7天。结论:除肝、肠、皮肤外,肺、舌可能也是GVHD靶器官。肝、脾组织中供体淋巴细胞浸润伴随MIP-1α水平的升高。This study was aimed to investigate the migration and distribution processes of allogeneic donor T lymphocytes in the organs of recipient mice, GVHD model was established by transfusion of the splenocytes of eGFP transgeneic C57BL/6 mice together with born marrow cells harvested from C57BL/6 mice into BALB/c mice underwent 8, 0 Gy total body irradiation, The migration and homing of eGFP ^＋ cells were tracked by stereo-fluorescent microscopy or inverted fluorescent microscopy and flow cytometry. The enzyme linked immunosorbent assay （ELISA） was performed on supernatants from the tissue homogenates to detect the amount of MIP-1α. The results indicated that GVHD clinical manifestation and pathological changes of organs appeared on day 8 post transplantation, eGFP-positive donor T cells in recipient organs were observed by inverted fluorescence microscope in frozen section, or by stereo-fluorescence microscopy in living organs, such as liver, spleen, skin, lungs, bowels, and tongue. The highest expression of MIP-1α was on day 7 post transplantation in the liver （491.3 ±32.1 pg/ml）, and day 3 post transplantation in the spleen （881.5 ± 45.2 pg/ml） ,respectively （P 〈 0.05 ）. It is concluded that GVHD was induced by splenocytes of eGFP transgeneic C57BL/6 mice. eGFP^＋ cells in the organs can be observed by fluorescent microscopy. In this GVHD model, donor T cells proliferate and infiltrate in liver, skin, bowels, as well as lungs and tongue. MIP-1α may be in relation with the infiltration of T lymphocytes in liver and spleen.

关 键 词：小鼠GVHD模型 T淋巴细胞 增强型绿色荧光蛋白 巨噬细胞炎症蛋白 

分 类 号：R457.7[医药卫生—治疗学]