异基因造血干细胞移植后慢性移植物抗宿主病小鼠模型的建立和评价

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-2kd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20mg/(kg·d)×4d+环磷酰胺150mg/(kg·d)×2 d;方案2:白消安20mg/(kg·d)×4 d+环磷酰胺100mg/(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20mg/(kg·d)×4d-+环磷酰胺150mg/(kg·d)×2d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.

Establishment and evaluation of mouse model of chronic graft-versus-host disease after allogeneic hematopoietic stem cell transplantation

[Abstract] Objective To establish a mouse model of chronic graft versus host disease after allogeneic hematopoietic stem cell transplantation by use chemotherapy preconditioning?regimens. Methods The donor mouse was Balb/H-2kd , the recipients mouse was C57BL/6 H-2kb and the pretreatment program was different doses of chemotherapy drugs (BU+CTX) , by injected different spleen cells and bone marrow cells to establish the model of chronic graft versus host disease . Results Bu (20mg/kg.d) +CTX (150mg/kg.d), 2×107 bone marrow mononuclear cells +6.0×107 spleen mononuclear cells can form a high level of donor-recept mixed(P < 0.05).The occurrence time of graft versus host disease was more concentrated in the 14~90d.The control group and the experimental group had significant difference in the quality of the mouses(P < 0.05).The clinical scores were significantly increased in the large numbers of cells and high doses of chemotherapy preconditioning?regimens transplantation group(P < 0.05).The incidence of chronic graft versus host disease was significantly increased(P < 0.05). Conclusion 2×107 bone marrow mononuclear cells +6.0 ×107 spleen mononuclear cells and 2×107 bone marrow mononuclear cells +4.0×107 spleen mononuclear cells can form a stable chronic graft versus host disease model which give a experiment basement about clinical treatment of ?chronic graft versus host disease by Pathologic score and clinical manifestation.