目的:建立小鼠非清髓异基因骨髓干细胞移植模型,探讨不同的供者淋巴细胞输注(donor lymphocytes infusion DLI)对移植效果的影响及其机制。方法:受者鼠为Balb/c小鼠,供者鼠为C57b1/6小鼠,依据不同DLI将受者鼠随机分为A、B、C、D和E5组,每组5只。5组均给予预处理,A组预处理后于移植d1给予供者小鼠全脾细胞1.0×10^7个;B组分别于移植后d1、7、14给予供者小鼠全脾细胞0.3×10^7个;C组于移植后d1、7给予去除CD8^8细胞供者鼠脾细胞05×10^7个;D组于移植后d1、7给予供者鼠全脾细胞0.5×10^7个,E组为对照组,只作预处理,不作干细胞移植及DLI处理。观察各组嵌合状态、造血恢复情况、移植物抗宿主反应(graft versus host disease GVHD)。结果:未移植组嵌合率为22.33%,为非特异性粘着,移植组小鼠嵌合率均在80%以上,与未移植组相比差别显著;除C组一只小鼠死亡外其余均存活,并于移植后20天自行恢复造血功能,但C组白细胞计数小于2.0×10^9/L持续到d16,恢复明显比其余组慢,GVHD评分A、B组间无明显统计学差异(P=0.173),C、D组间相比有统计学差异(P=0.033)。结论:建立非清髓小鼠异基因骨髓干细胞移植模型条件适宜,且去除CD8^+细胞的DLI移植后GVHD比未去除组要严重,移植效果相对较差。
Objective:Establish a mouse model of allo-BMT with nonlethal preconditioning, observe the effect and research mechanism of allo -BMT with different DLI . Methods: Recipient mouse was Balb/c and donor was C57bl/6. Twenty five recipient mice were divided into A,B,C,D and E group according to different DLI randomly. All the mice were preconditioned, the mice in group A were treated with 1.0×10^7 donor mice's whole spleen cells on day+1 after transplantation ; The mice in group B were treated with 0.3×10^7 donor mice's whole spleen cells on day+1, +7, +14 after transplantation; The mice in Group C were treated with 0.5×10^7 donor mice's CD8-spleen cells on day+1, +7 after transplantation; The mice in group D were treated with 0.5×10^7 donor mice's whole spleen cells on day+1, +7 ; Group E , the control group, was just preconditioned. Then the chimerism, hematopoiesis recovery and GVHD were observed. Results: The chimefism of group A,B,C and D were above 80% ,superior to that of control group. All the mice survived except one died of GVHD in group C , while the hematopoiesis recovery of group C was slower than the others. There was no statistic difference between group A and B in GVHD scores(P=0.173 ), however difference existed between group C and D (P=0.033 ). Conclusion :The results indicated that the nonlethal precondition of the mouse model is suitabl; The mice that treated with CD8-DLI showed more severe GVHD than that with whole spleen cell DLI.