单倍体相合造血干细胞移植急性移植物抗宿主病小鼠模型的构建和发病机制的探讨

发布时间：2018-04-03 21:08

本文选题：半相合造血干细胞移植　切入点：急性移植物抗宿主病　出处：《中国实验血液学杂志》2017年04期

【摘要】：目的:构建单倍体相合造血干细胞移植(HSCT)小鼠急性移植物抗宿主病(aGVHD)模型,并探讨其发病机制。方法:建立以C57BL/6(H-2~b)为供鼠,137Csγ射线11 Gy照射[C57BL/6×CBA/Ca]F1(H-2b×k)为受鼠的单倍体相合造血干细胞移植aGVHD模型。将受鼠随机分为照射对照、去除T细胞的骨髓细胞(TD-BM)、TD-BM+T细胞(1)和TD-BM+T细胞(2)共4组。移植后对受鼠生存期和体重进行评分;比较移植后14 d受鼠的靶器官损伤情况,比较TD-BM和TD-BM+T细胞(1)组的病理评分;比较移植后14和28 d TD-BM和TD-BM+T细胞(1)组的骨髓嵌合率;应用ELISA法分别检测移植后14 d TD-BM和TD-BM+T细胞(1)组受鼠血清中细胞因子IFN-γ、TNF-ɑ和IL-1的浓度。应用流式细胞术检测TD-BM和TD-BM+T细胞(1)组受鼠脾细胞中H-2b+H-2k-CD4+IFN-γ+Th1细胞的百分比和CFSE检测供者T细胞发生的增殖情况。结果:TD-BM、照射对照、TD-BM+T细胞(1)和TD-BM+T(2)组平均活存时间分别为60、13.29±5.50、33±2.35、29.14±1.77 d。移植后14 d,TD-BM+T细胞(1)和TD-BM+T(2)组小鼠出现肝脏、结肠、肺脏和皮肤等多器官损伤,而在TD-BM组多器官损伤不明显。移植后14和28 d,TD-BM和TD-BM+T细胞(1)组小鼠的嵌合率均94%。移植后14 d,TD-BM+T细胞(1)受鼠血清中细胞因子IFN-γ、TNF-ɑ和IL-1水平较TD-BM组明显增高。移植后14 d,TD-BM+T细胞(1)组中H-2b+H-2kCD4+IFN-γ+百分比明显高于TD-BM组[(0.5240±0.08447)%vs(7.912±0.6087)%](P0.05)。移植后14d,供者T细胞在受者体内发生明显增殖。结论:C57BL/6(H-2b)→[C57BL/6×CBA/Ca]F1(H-2b×k)可作为单倍体相合移植aGVHD的实验模型;Th1细胞在单倍体相合移植aGVHD中可能发挥重要作用。
[Abstract]:Aim: to establish a model of acute graft-versus-host disease (GV HDD) in haploid compatible hematopoietic stem cell transplantation (HSCT) mice and to explore its pathogenesis.Methods: the aGVHD model of haploid compatible hematopoietic stem cell transplantation (HSCT) was established using C57BL / 6H ~ (-2) B) as the donor of 137Cs 纬 -ray [C57BL/6 脳 CBA/Ca] F1(H-2b 脳 kY.The mice were randomly divided into 4 groups: bone marrow cells (TD-BMN, TD-BM T cells 1) and TD-BM T cells (2).The survival time and body weight of the recipient rats were evaluated after transplantation, the target organ injury was compared 14 days after transplantation, the pathological score of TD-BM and TD-BM T cell group was compared, and the bone marrow chimerism rate of TD-BM and TD-BM T cell group was compared 14 and 28 days after transplantation.The levels of cytokines IFN- 纬 TNF- and IL-1 in the serum of TD-BM and TD-BM T cell group were detected by ELISA method on the 14th day after transplantation.The percentage of H-2b H-2k-CD4 IFN- 纬 Th1 cells in recipient spleen cells and the proliferation of donor T cells in TD-BM and TD-BM T cells were detected by flow cytometry.Results the mean survival time of TD-BM T cells in the control group (TD-BM1) and TD-BM Tn2) group was 60 ~ 13.29 卤5.50 ~ (33 卤2.35) ~ (29.14 卤1.77) days, respectively.On the 14th day after transplantation, liver, colon, lung and skin were damaged in mice with TD-BM T cell 1) and TD-BM Tn2), but not in TD-BM group.The chimerism rates of TD-BM and TD-BM T cells on 14 and 28 days after transplantation were 94.On the 14th day after transplantation, the levels of cytokines IFN- 纬 TNF- and IL-1 in the serum of TD-BM T cells were significantly higher than those in the TD-BM group.14 days after transplantation, the percentage of H-2b H-2kCD4 IFN- 纬 in the TD-BM T cell group was significantly higher than that in the TD-BM group [0.5240 卤0.6087 卤0.6087%].On the 14th day after transplantation, the donor T cells proliferated obviously in the recipient.Conclusion [C57BL/6 脳 CBA/Ca] F1(H-2b 脳 k] [C57BL/6 脳 CBA/Ca] F1(H-2b 脳 k) can be used as an experimental model of haploid aGVHD transplantation. Th1 cells may play an important role in haploid aGVHD transplantation.
【作者单位】：武汉大学中南医院血液科;昆明医科大学第二附属医院血液科;
【基金】：国家自然科学基金青年基金(81500151);国家自然科学基金地区基金(81160074) 武汉大学自主科研青年教师项目(2042014kf0167)
【分类号】：R-332;R457.7

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