异基因骨髓细胞在致敏模型体内归巢示踪与植入分析  

Homing and engraftment of allogeneic bone marrow cells in a sensitized mouse model

作　　者：许吕宏[1] 方建培[1] 翁文骏[1] 徐宏贵[1] 

机构地区：[1]中山大学孙逸仙纪念医院儿科,广东广州510120

出　　处：《中国病理生理杂志》2011年第8期1457-1461,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30872383);广东省医学科研基金资助项目(No.B2010086);广东省自然科学基金资助项目(No.9151008901000111);高等学校博士学科点专项科研基金资助项目(No.20090171110064)

摘　　要：目的:探讨异基因供者骨髓细胞在致敏模型体内的归巢示踪与植入分析。方法:以异基因脾细胞输注方法建立致敏的BALB/c小鼠模型,同时取正常BABL/c小鼠作为非致敏模型。致敏或非致敏模型经8 Gy[60Co]照射后分别经尾静脉移植1×107 C57BL/6小鼠骨髓细胞。用绿色荧光染料CFSE标记供者骨髓细胞,并分别在移植后不同时点(2 h、12 h及48 h),通过组织细胞悬液动态示踪供者细胞在致敏受者各组织的分布。移植后记录各组的生存情况,每周监测造血重建与骨髓恢复情况。予H-2Db进行标记移植后受者骨髓细胞,检测供者嵌合百分比。结果:CFSE能标记供者骨髓细胞并用于体内示踪实验。动物体内归巢示踪实验表明,与非致敏组相比,异基因供者骨髓细胞在致敏受者体内的外周血、脾脏及股骨的分布均明显减少。植入分析结果发现,非致敏受者于移植后能长期存活,外周血及骨髓细胞均能迅速恢复;而致敏组中,小鼠均于移植后2周左右全部死亡,生存中位数为13 d,外周血及骨髓细胞均随时间推移呈进行性减少。嵌合分析显示移植后第7 d,非致敏受者与致敏受者的供者骨髓细胞百分比分别为(48.07±4.70)%和(0.77±0.11)%,两者差异显著(P<0.01)。结论:异基因供者骨髓细胞在致敏受者体内脾脏及股骨等部位被清除,不能有效植入。AIM： To investigate homing and engraftment of allogeneic bone marrow ceils （BMCs） in a sensitized mouse model. METHODS： Sensitized BALB/c mice were established by allogeneic splenocytes transfusion. Normal BALB/c mice were used as non - sensitized controls. After 8 Gy of [ ^60Co ] irradiation, the sensitized or non - sensitized mice received 1×10^7 BMCs from C57BL/6 donor intravenously. A homing assay was also performed by labeling the donor cells with the cell tracker CFSE. Cell suspensions from different tissues were analyzed at different time points （2 h, 12 h and 48 h after transplantation）. The survivals were assessed after transplantation, and hematopoietic recovery was monitored. BMCs from recipients were labeled with H -2Db and the percentage of donor cells was calculated. RESULTS： BMCs labeled with CFSE was suitable for homing study in vivo. Compared with the non - sensitized recipients, the homing assay showed allogeneic BMCs significantly decreased in peripheral blood, spleen and femur of sensitized recipients. Long - term survival was observed in non - sensitized recipients after transplantation by engraftment study, and hematopoiesis recovered rapidly along with time. However, all the sensitized recipients died within 2 weeks after transplantation with a median of 13 days, and hematopoiesis decreased along with time. Severn days aftertransplantation, the percentages of allo- geneic BMCs in non -sensitized and sensitized recipients were （48.07±4.70） % and （0. 77 ±0. 11 ） %, respectively, with statistically significant difference （P 〈 0. 01 ）. CONCLUSION： Allogeneic BMCs are depleted in spleen and femur of sensitized recipients, and could not engraft in the sensitized recipients.

关 键 词：骨髓细胞 致敏模型 归巢 植入 

分 类 号：R457[医药卫生—治疗学]