同基因、异基因骨髓移植小鼠模型中T淋巴细胞组织浸润的观察  

Infiltration of T lymphocytes in allogeneic and autogenous bone marrow transplantation murine model

作　　者：温宏升[1] 王健民[1] 齐海燕[1] 周红[1] 李津英[1] 夏荣[1] 邱慧颖[1] 

机构地区：[1]第二军医大学长海医院血液科,上海200433

出　　处：《第二军医大学学报》2005年第7期779-782,共4页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金(39870710;30172347);上海市卫生系统百名跨世纪优秀学科带头人培养基金(98BR029).

摘　　要：目的:观察异基因骨髓移植中,供体T淋巴细胞在受体各器官(包括肝、肠、皮肤、肺、肾、脑和心肌等)中的分布情况,以及与同基因移植的异同。方法:以尼龙毛法从转增强型绿色荧光蛋白(eGFP)基因C57BL/6小鼠和C57BL/6小鼠脾细胞中分离T淋巴细胞,以红色荧光细胞膜染料PKH26(10μmol/ml)染色C57BL/6小鼠T淋巴细胞,将eGFP+T淋巴细胞或PKH26+T淋巴细胞与C57BL/6小鼠骨髓细胞一起经尾静脉分别输入经8.0Gy全身照射的同基因受体小鼠(C57BL/6)和异基因受体小鼠(BALB/c)体内,建立小鼠同基因、异基因骨髓移植模型。在移植后第1天、第4天、第8天处死小鼠,观察肝、肠、皮肤、肺、肾、心、脑等组织冰冻切片中荧光细胞的分布。结果:异基因骨髓移植模型中,移植后第1天,受体小鼠各器官中未见绿色或红色荧光细胞;移植后第4天、第8天可见绿色或红色荧光细胞,主要分布于肝、肠、皮肤、肺,而肾、心、脑等组织中未见;移植后第8天肝、肠、皮肤、肺中eGFP+T淋巴细胞明显多于第4天;而移植后第8天肝、肠、皮肤、肺中PKH26+T淋巴细胞明显少于第4天(细胞膜上PKH26因细胞分裂而逐渐减少)。同基因骨髓移植模型中,移植后第1天、第4天和第8天,肝、肠、皮肤、肺、肾、心、脑组织中均未见红色或绿色荧光细胞。结论:异基因骨髓移植中,可能因供体T淋巴细胞被激活、增殖,故可大量进入受体肝、肠、皮肤、肺等器官;肝、肠、皮肤是公认的移植物抗宿主病(GVHD)靶器官,肺可能也是潜在的GVHD靶器官。同基因骨髓移植中,可能因供体T淋巴细胞不能被激活、增殖,因而不能大量进入受体肝、肠、皮肤、肺等器官。Objective: To investigate the distributive difference of donors’ T lymphocytes in the recipients’ organs, including the liver, bowel, skin, lungs, brain, kidneys, and heart between allogeneic and autogenous bone marrow transplantation (BMT) animal models. Methods: T lymphocytes were isolated from splenocytes of C57BL/6 mice and C57BL/6 mice transfected with enhanced green fluorescent protein (eGFP) by the nylon adhering method. The T lymphocytes of C57BL/6 mice were dyed with PKH26 (a kind of cellular membrane dye with red fluorescence).The autogenous BMT model was infused through vena caudalis with eGFP+T lymphocytes ( 4×106) plus C57BL/6 born marrow cells (8×106) after fatal dose total body irradiation (TBI) 8.0 Gy, and the allogeneic BMT model with PKH26-dyed T lymphocytes (4×106) plus C57BL/6 born marrow cells (8×106). The distribution of T lymphocytes from the donor was observed in the frozen sections of the liver, bowel, skin, lung, kidney, brain and heart on days 1, 4 and 8 post BMT with an inverted fluorescent microscope (Olympus Ⅸ70). Results: In the Allo-BMT model, eGFP+cells and PHK26+cells were seen in the liver, bowel, skin and lung on days 4 and 8 post-transplantation, but not in the kidney, brain and heart. More eGFP+cells were seen on day 8 than on day 4, and fewer PHK26+cells were seen on day 8 than on day 4 (PKH26 on cellular membrane tapered with cell division). No eGFP+cells or PHK26+cells were seen on day 1 in any organ in the Allo-BMT model. In the Auto-BMT model, no eGFP+or PKH26+cells were seen in any organ on days 1, 4 or 8. Conclusion: In the Allo-BMT model, donor T lymphocytes can be activated, proliferate and migrate into the liver, intestine, skin and the lungs. The liver, skin and intestine are the classic GVHD target organs and lungs may be a potential GVHD target organ. T lymphocytes can not migrate into the organs of the recipient in the Auto-BMT model because the donor's T lymphocytes can not be activated.

关 键 词：骨髓移植 T淋巴细胞 增强型绿色荧光蛋白 PKH26 

分 类 号：R457.7[医药卫生—治疗学]