输注脾细胞建立输血相关的移植物抗宿主模型  

Development of transfusion associated graft vs host disease model by allogeneic splenocytes infusion

作　　者：陈必成[1] 昌盛[1] 向芙莉[1] 郭晖[1] 唐莉[1] 金昊[1] 曾宁[1] 陈忠华 

机构地区：[1]华中科技大学同济医学院附属同济医院器官移植研究院

出　　处：《中国输血杂志》2006年第1期2-4,共3页Chinese Journal of Blood Transfusion

基　　金：国家自然科学基金资助课题(编号:30271243);国家重点基础研究发展规划项目(编号:2003CB515505);教育部/卫生部器官移植国家重点实验室

摘　　要：目的探讨输注同种脾细胞建立小鼠输血相关的移植物抗宿主(TA-GVHD)模型及其特点。方法分离C57BL/6(H-2b)脾细胞,分别经静脉输注给Balb/c(H-2d)裸鼠与F1代(C57BL/6×Balb/c H-2b/d),建立输血相关的移植物抗宿主模型。活体染料羧基荧光素乙酰乙酸(5,6-carboxyfluorescein diacetate succini midyl ester,CFSE)标记供者脾细胞,通过流式细胞仪分析供者细胞在受者体内的增殖;免疫组化分析供者脾细胞survivin的表达;检测Balb/c裸鼠体内抗供者抗体及IL-10浓度;分析皮肤、小肠、肝脏的病理变化。结果输注同种脾细胞成功建立急性TA-GVHD模型。受者脾脏及外周可以检测到标记后增殖的供者细胞。小鼠的病理改变主要出现在肝脏,浸润细胞早期表达survivin。小肠及皮肤未见明显的病理改变。TA-GVHD裸鼠第7天IL-10、抗-Balb/c显著升高。结论输注同种脾细胞可以建立TA-GVHD模型,并具有简单、快速、稳定的特性,为诊断、治疗GVHD提供了良好的模型。Objective To develop transfusion associated graft vs host disease（TA-GVHD） model by infusing allogeneic splenocytes. Methods C57BL/6 （H-2^b） splenocytes were harvested, labeled by 5,6-carboxyfluorescein diacetare succinimidyl ester （CFSE）, and infused into Balb/c nude mice （H-2^d） and FI mice （C57BL/6 × Balb/c, H-2^b/d） to develope TA-GVHD model. Labeled donor splenocytes were detected by flow cytometry. Survivin expression of donor splenocytes in host liver was detected by immunohistochemistry. Anti-donor antibody and IL-10 in serum of Balb/ c nude mice were measured by complement-dependent cytotoxicity and ELISA, respectively. Results TA-GVHD could be developed by infusing allogeneic splenocytes. Donor cells were detected in the spleen and peripheral blood of recipient. Liver injury was serious but no injury in the skin and the intestine was observed. The expression of survivin in lymphocytes was detected in the liver of FI and nude mice. TA-GVHD nude mice had high concentrations of IL-10 and anti-Balb/c antibodies after splenocyte infusion. Conclusion TA-GVHD model can be established in nude Balb/ c and F1 mice by infusing allogeneic splenocytes.

关 键 词：移植物抗宿主病 输血相关 输注/脾细胞TA-CVHD模型/棵鼠 淋巴细胞 

分 类 号：R457.13[医药卫生—治疗学]