单倍体供者淋巴细胞输注诱导慢性移植物抗宿主病小鼠模型的建立
Construction of a murine model of chronic graft-versus-host disease by haplo-identical spleen cell infusion
摘要目的 建立单倍体相合供者淋巴细胞输注(DLI)后慢性移植物抗宿主病(cGVHD)小鼠模型并对其进行鉴定.方法 以近交系雄性Balb/cH-2d小鼠为供鼠;以雌性CB6F1小鼠(Balb/c×C57BL/6 F1H-2d/b)为受鼠,随机分为移植3×107、6×107、9×107个脾细胞组及空白对照组,脾细胞移植后流式细胞学检测H-2b和H-2d进行嵌合体分析.观察受鼠的体重、体形、体位、毛发变化及生存情况并评分,移植100 d处死小鼠,观察皮肤、肝脏、回盲端小肠靶器官的病理变化并评分,并鉴定模型的特征.结果 输注6×107、9×107个Balb/c小鼠脾细胞组可较为稳定的诱导小鼠cGVHD模型,两组间cGVHD的发生率的差异无统计学意义(P>0.05),统计学分析提示是以皮肤硬皮病样改变的小鼠慢性GVHD模型.结论 输注6×107、9×107个Balb/c供鼠脾细胞的CB6F1受鼠可建立以皮肤硬皮病改变为主的稳定的cGVHD小鼠模型.
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abstractsObjective For providing experimental platform of chronic graft-versus-host disease (cGVHD),to establish a mouse model by haplo-identical spleen cell infusion.Methods The donor male mice (Balb/cH-2d) and the recipient (Balb/c C57BL/6) F1 H2-d/b (CB6F1) female mice were randomly divided into four groups:3 experimental groups injected with 3 107,6 107 and 9 107 spleen cells,respectively,while the control group received RPMI 1640 solution.H-2d and H-2b were checked to analyze the chimerism in bone marrow cells.Body mass,figure,cutaneous manifestation and survival of recipient mice were observed and scored every 3 days.Pathologic changes of target organs were observed and scored.Results Injection of 6 107 and 6 107 splenocytes in the recipient mice resulted in a chronic disease with a low level of parental cell engraftment steadily.As compared with 3 107 group,the incidence of cGVHD in 6 107 and 9 107 groups were significantly increased (P <0.01).But there was no significant difference between 6 107 and 9 107 groups (P>0.05).Conclusion A murine model of cGVHD after haplo-identical spleen cell infusion of donor is successfully established by injection of 6 107 and 9 107 spleen cells.
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