| 摘要: |
| 目的 采用化疗药物预处理方案建立异基因造血干细胞移植的慢性移植物抗宿主病小鼠模型，并进行评价。方法 以BALB/cH-2kd小鼠作为供鼠，C57BL/6H-2kb小鼠为受鼠。对受鼠使用不同的化疗药物进行预处理[方案1：白消安20 mg/（kg·d）×4 d+环磷酰胺150 mg/（kg·d）×2 d；方案2：白消安20 mg/（kg·d）×4 d +环磷酰胺100 mg/（kg·d）×2 d]，然后经尾静脉注射不同剂量的供鼠脾细胞（6×107或4×107个）和（或）相同剂量的骨髓细胞（2×107个），建立慢性移植物抗宿主病小鼠模型；采用嵌合体分析、临床评分、组织病理学等进行评价。结果 采用白消安20 mg/（kg·d）×4 d +环磷酰胺150 mg/（kg·d）×2 d的方案进行预处理、2×107个骨髓单个核细胞+6×107个脾单个核细胞进行移植可形成较高水平的供受者混合嵌合体。移植物抗宿主病发生时间多集中在供鼠脾细胞输注后 30~90 d，化疗药物剂量大的预处理方案及移植细胞数高的移植组小鼠的临床评分和慢性移植物抗宿主病的发生率高于化疗药物剂量小的预处理方案及移植细胞数少的移植组（P<0.05）。模型小鼠肠、肝脏、皮肤、脾脏等器官出现细胞和结构异常、炎症细胞浸润等病理改变。结论 采用白消安和环磷酰胺预处理、给予2×107个骨髓单个核细胞+6×107或4×107个脾单个核细胞可形成较稳定的慢性移植物抗宿主病小鼠模型，为进一步指导临床治疗慢性移植物抗宿主病奠定了实验基础。 |
| 关键词: 动物模型 移植物抗宿主病 同种移植 造血干细胞移植 |
| DOI：10.16781/j.0258-879x.2016.12.1501 |
| 投稿时间：2016-07-17修订日期：2016-11-25 |
| 基金项目:珠海市医学科研基金（2015J030）. |
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| Establishment and evaluation of mouse model of chronic graft-versus-host disease after allogeneic hematopoietic stem cell transplantation |
| YI Wen-fang1*,GUO Kun-yuan2,HE Xin2,LIN Guang1,ZHANG Cheng1,WANG Jing2 |
(1. Department of Pediatrics, Zhuhai People's Hospital, Zhuhai 519000, Guangdong, China;
2. Department of Hematology, Zhujiang Hospital, Southern Medical University, Guangzhou 510280, Guangdong, China
* Corresponding author) |
| Abstract: |
| Objective To establish a mouse model of chronic graft-versus-host disease (GVHD) after allogeneic hematopoietic stem cell transplantation using chemotherapy preconditioning regimens. Methods The donor mouse was BALB/cH-2kd and the recipients mouse was C57BL/6H-2kb. The pretreatment of recipient mouse was performed using different doses of chemotherapy drugs, busulfan (BU) 20 mg/(kg·d)×4 d+cyclophosphamide (CTX) 150 mg/(kg·d)×2 d, or BU 20 mg/(kg·d)×4 d+CTX 100 mg/(kg·d)×2 d, and then the recipient mouse was injected with different doses of spleen cells (6×107 or 4×107 cells) and/or same dose of bone marrow cells (2×107 cells) to establish the model of chronic GVHD. Hematopoietic chimerism analysis, clinical score and histopathology were used to evaluate the model. Results The high level of recipient-donor mixture chimera was established by pretreated by BU 20 mg/(kg·d)×4 d+CTX 150 mg/(kg·d)×2 d and 2×107 bone marrow mononuclear cells +6×107 spleen mononuclear cells. The onset of GVHD was concentrated at the 30-90 d after injecting spleen cells of the donor mouse. The clinical scores and incidence rate of chronic GVHD were significantly increased in the large numbers of cells and high doses of chemotherapy transplantation group compared with those in smaller numbers of cells and lower doses of chemotherapy groups (P<0.05). Pathological changes such as cellular and structural abnormalities and inflammatory cell infiltration were noted in the intestines, liver, skin and spleen of the model mice. Conclusion Bone marrow mononuclear cells (2×107 cells)+spleen mononuclear cells (6×107 or 4×107 cells) can be used to establish a stable chronic GVHD mouse model via preconditioning with BU and CTX, which paves a way for clinical treatment of chronic GVHD. |
| Key words: animal models graft vs host disease homologous transplantation hematopoietic stem cell transplantation |
