小鼠脾细胞在混合骨髓移植中的作用

Effects of the splenocytes in murine mixed bone marrow transplantation

目的:观察同异基因小鼠脾细胞输注比例和输注量在混合半相合骨髓移植中对移植物抗宿主病(GVHD)和嵌合状态的影响。方法:受鼠全身照射后,经尾静脉回输同基因和半相合异基因供鼠骨髓及两者不同比例脾细胞。A组未加入异基因供鼠脾细胞,B组加入的同异基因供鼠脾细胞量相等,C组加入异基因供鼠脾细胞量是同基因供鼠脾细胞量的两倍。回输后观察GVHD临床表现,取肝、皮肤、小肠送病理学检查评定GVHD分级。测定+70天、+150天嵌合率。监测造血重建和受鼠生存率。结果:混合骨髓移植各组小鼠生存期均超过50天。其中A组,临床和病理无GVHD表现,+150天嵌合水平大幅度下降。B组,临床和病理无明显GVHD表现,+150天嵌合水平中等幅度下降。C组病理有GVHDⅡ级表现,+150天嵌合水平下降不明显。半相合骨髓移植对照组小鼠出现典型的GVHD临床和病理表现,仅20%小鼠生存期超过50天。结论:小鼠混合半相合骨髓移植中,同异基因小鼠脾细胞输注比例和输注量对移植排斥和GVHD有协调作用。

Objective：To explore the effects of splenocytes on graft-versus-host disease （GVHD） and chimerism in murine mixed bone marrow transplantation. Methods：During mixed transplantation, syngeneic bone marrow cells from BALB/c mice were mixed with H-2 haploidentical matched bone marrow cells from CB6F1 mice. At the same time, different infusion dose of splenocytes were transplanted into lethally irradiated groups of BALB/c mice. BALB/c mice of group A hadn＇t accepted splenocytes of CB6F1 mice. BALB/c mice of group B were infused with the same dose of syngeneic and CB6F1 mice splenocytes. BALB/c mice of group C accepted the double dose of splenocytes of CB6F1 mice compared with syngeneic splenocytes. After transplantation, the appearance of a GVHD, the level of the chimerism of the recipient mice were observed and detected by histopathological examination on liver, skin, small intestine and flow cytometry. Hematopoietic recovery and the mortality were evaluated by blood routine test and clinical observation. Results：Mice in mixed transplantation groups survived over 50 days. No GVHD manifestation and histopathological alterations appeared in group A, and the chimerism on the 150th day decreased significantly. Mice of group B showed little GVHD manifestation and histopathological changes, and the chimerism on the 150th day was reduced. Mice of group C showed a higher level and more stable of chimerism with grade Ⅱ degree of GVHD histopathological aherations. Typical GVHD manifestation and histopathological alterations appeared in H-2 haploidentical bone marrow transplantation group. The survival rate was only 20% 50 days after transplantation. Conclusion ：The proportion and dose of syngeneic and haploidentical MHC splenocytes were of importance to balance GVHD and host-versus-graft reaction（HVGR）.