同种异体骨移植后小鼠脾脏T淋巴细胞亚群的变化

Spleen T cell subsets observation post mice allogeneic bone graft

目的 通过植入不同方法处理的同种异体骨 ,观察移植免疫排斥中脾脏T细胞的总数与亚群的变化 ,探讨同种异体骨移植后机体的免疫反应。方法 取 64只C5 7BL/6、1 6只BALB/C小鼠的双侧肱骨、股骨及胫骨 ,制成新鲜异体骨、冷冻干燥异体骨、冷冻干燥脱脂异体骨、脱脂脱钙异体骨四种同种异体骨 ,及新鲜自体骨 ;每只小鼠骨对等分为 2份 ,分别植入 1 60只BALB/C小鼠双侧股四头肌囊袋内。于术后 1、2、4、8周处死 ,提取脾脏T淋巴细胞。用FITC特异性荧光标记的CD3、CD4、CD8单克隆抗体作用后 ,FACS流式细胞仪检测 ,得到T淋巴细胞及其亚群百分数。结果 接受C5 7BL/6同种异体骨移植后BALB/C小鼠T细胞总数及亚群于术后 1周、2周渐升高 ,4周开始下降 ,8周基本恢复正常。其中新鲜异体骨移植组显著高于其它各组 (P <0 .0 0 5 ) ,冷冻干燥异体骨、冷冻干燥脱脂异体骨、脱脂脱钙异体骨之间的差异无统计学意义 (P >0 .0 5 )。不同时间点的差异有统计学意义 (P <0 .0 0 5 )。结论 冷冻干燥异体骨、冷冻干燥脱脂异体骨、脱脂脱钙异体骨移植后的细胞免疫水平相似。

Objective In order to look further into the mechanism of immune rejection of allogeneic bone graft,the immune reaction about T lymphocyte subsets between 4 kinds of different allogeneic bone were compared.Methods 64 C57BL/6 and 16 BALB/C mice were humanly sacrificed,then bilateral humeral,femur and tibia were taken out. These bones were dealt with different methods,and be made into fresh bone,deeply freeze-dried bone,fat-extracted bone,and defatted-demineralized bone. Then they were equally put into the thigh muscle pocket of 160 BALB/C mice. By the time of 1,2,4,8 weeks postoperative,the mice were killed,and the spleens were picked out. Spleens T lymphocytes were collected up. Then monoclonal antibodies were added into the dilution,which included CD3,CD4,and CD8 labeled with FITC. The positive rates of cells combined with monoclonal antibodies were counted in FACSan. Results The rates of CD3,CD4,CD8 T-cells ascended in 2 weeks postoperative,descended by the time of 4 weeks,and reached normal by 8 weeks. By the time of 2～4 weeks postoperative,the indexes of fresh allogeneic bone were higher significantly than that of the other groups(P<0.005); while there were no differences between the indexes of freeze-dried bone,fat-extracted bone,and defatted-demineralized bone(P>0.05). The indexes were significantly variable due to different time post operation (P<0.005). Conclusion The immune rejections of fresh alloneneic bone graft were significant,while that of freeze-dried bone,fat-extracted bone,and defatted-demineralized bone were somewhat at the same level. Trauma affected the indexes significantly.