同种异基因小鼠骨髓移植后脾细胞的动力学研究

本研究建立同种异基因小鼠骨髓移植模型，监测移植后脾细胞的动力学变化，了解移植后外周免疫器官的免疫重建。供鼠为C57BL／6小鼠，受鼠为BALB／c小鼠。将受鼠分为单纯照射组（R组）、照射＋输注骨髓单个核细胞组（B组）、照射＋输骨髓单个核细胞＋输脾单个核细胞组（S组）。移植后观察小鼠体重、体位、活动性、毛发、腹泻及生存期，每周检测外周血白细胞及血红蛋白，于移植后第2、7、14、27、60天计数S组小鼠脾脏单个核细胞数，检测S组小鼠脾脏单个核细胞中CD3^＋、CD4^＋、CD8^＋、B220^＋及CD11c^＋细胞的相对计数，濒死前行肝、小肠、皮肤病理检查。结果显示：S组小鼠中89％于移植后第6—78天死于aGVHD。移植后脾脏单个核细胞迅速下降，于移植后第2天降至最低点，持续至第7天，之后逐渐回升，第14天升至移植前水平；CD8^＋细胞和B220^＋细胞于移植后第2天基本降至最低点，CD8^＋细胞恢复最快，在第7天接近移植前水平，且大多为供者型，而B220^＋细胞恢复最慢，从第2—14天持续低水平，之后逐渐回升，但直至第60天仍未恢复至移植前水平；CD3^＋细胞和CD4^＋细胞降低缓慢，于第7天开始出现少量供者型细胞，总的比例逐渐下降，至14天降至最低点，之后逐渐回升，第60天时均恢复至移植前水平；CD11c^＋细胞移植后除第14天升高外，其它时间点比例变化不明显，也是由受者型逐渐向供者型转化。结论：C57BL／6为供体，BALB／c为受体并给予7．5Gy^60 Coγ清髓的预处理条件下，输注骨髓单个核细胞1×10^7，脾单个核细胞1×10^7建立同种异基因移植模型，移植后脾细胞呈明显的动力学变化，CD8^＋细胞重建最快．茸次为CD3^＋、CD4^＋细胞．B220^＋细胞重建最慢。

Kinetic Study of Splenocytes after Allogeneic Murine Bone Marrow Transplantation

The study was purposed to understand immunological reconstitution of peripheral immune organs after transplantation, through establishing allogeneic murine bone marrow transplantation model and detecting the kinetic change of splenocytes after transplantation. C57BL/6 mice were donors, BALB/c mice were recipients. Recipient mice were divided into irradiation group（ R）, irradiation plus inoculating bone marrow mononuclear cells（MNC） group（ B ）, and irradiation plus inoculating bone marrow mononuclear cells and spleno-MNC group （ S ）. After transplantation, the mice were examined daily for the symptoms such as weight, hunched posture, activity, ruffled fur, diarrhea, and survival. Blood routine test was done once a week, splenocyte was counted and CD3, CD4, CD8, B220, CD11 c postive cell relative count was detected by FACS on day 2, 7, 14, 27, 60 after transplantation, Liver, skin and intestine were biopsied for histopathological examination before dying. The results indicated that 89% mice in S group died of acute graft-versus-host disease （aGVHD） during day 6 to 78. The spleno-mononuclear cell count quickly decreased and reached to lowest level on day 2, then gradually recovered to level of pretransplantation on day 14; CD8 and B220 positive cells decreased to lowest level on day 12, in which CD8 ~ cells quickly recovered and reached to level of pretransplantation, but the B220 ~ recovered most slowly and sustained to be with low level, then gradually recovered to level of pretransplantation on day 60; CD3 and CD4 positive cells decreased relatively slowly, and reached to lowest level on day 14, then both gradually recovered to level of pretransplatation on day 60; CDllc positive cell count changed unstrinkingly except day 14. It is concluded that when C57BL/6 mice are donors, and BALB/c mice are recipients treated with irradiation of 7.5 Gy and inoculated with 1 x 10^7 bone marrow MNC and 1 x 10^7 spleno-MNC, allogeneic murine bone marrow transplantation model can be thus set up. The spleno-mononuclear cells show obviously kinetic changes after transplantation, that CD8 positive cells recover ahead of all, then followed by CD3 and CD4 positive ceils, and the reconstitution of B220 positive cells is the slowest.