小鼠单倍体相合外周血移植模型的建立

目的建立单倍体相合外周血移植小鼠模型,探讨移植后造血及免疫重建特性。方法经大剂量全身照射预处理BDF1小鼠后,经尾静脉输注新生第一天DBA/2小鼠的外周全血,移植后动态观察受体鼠的造血重建及急性移植物抗宿主病(GVHD)发生情况,并用流式双染色法进一步检测粒系、淋巴系造血细胞的长期植入水平。结果移植后小鼠的白细胞、血小板、血红蛋白分别在第18、21、18天明显回升,其中白细胞和血小板分别于第29天和36天恢复正常,而血红蛋白至第50天仍为部分恢复。受体鼠90%长期存活,且不发生明显的GVHD。移植后第50天流式细胞仪检测供鼠来源的造血细胞占总有核细胞的88.81%,其中含供者来源的粒细胞33.92%,T细胞16.83%。结论单份新生小鼠外周血含有足够的造血干细胞,能重建单倍体相合小鼠的造血及免疫系统而不引起明显的GVHD,并可形成稳定的以供者细胞为主的嵌合体,其中粒细胞为完全嵌合。

Establishment of murine model of haploidentical peripheral blood transplantation

Objective To establish a murine model of haploidentical peripheral blood transplantation and evaluate the characteristics of hematopoietic and immunological recovery. Methods BDF1 recipient mice conditioned with high dose total body irradiation were transplanted with neonatal peripheral blood from DBA/2 mice through tail vein.Hematopoietic recovery and graft-versus-host disease(GVHD) were observed,and the engraftment level of neutrophils and T cells were further detected by double-color flow cytometry. Results After transplantation,the levels of white blood cell(WBC),platelet(PLT) and haemoglobin(Hb) were increased significantly on day 18,21 and 18,respectively.WBC and PLT recovered within normal range on day 29 and 36,respectively,while Hb had just been partially retrieved until day 50.Long term survival was achieved in 90% of the recipient mice,and there was no evidence of GVHD.It was revealed by flow cytometry on day 50 that the proportion of donor cells in nucleated cells of recipient peripheral blood was 88.1%,including 33.92% donor neutrophils and 16.83% donor T cells. Conclusion Each newborn peripheral blood unit yields enough progenitor cells to recover the hematopoietic and immunologic system in haploidentical mice without causing GVHD.The close to full donor type chimerism was achieved,and the neutrophil was totally engrafted.