小鼠同种异体及异种神经移植后脾脏T淋巴细胞亚群的变化

目的通过化学法萃取的去细胞异体及异种神经移植后,观察移植免疫排斥过程中小鼠脾脏T细胞总数与亚群变化,探讨周围神经缺损修复的可行性。方法BALB/c小鼠随机分成假手术组、新鲜自体神经移植组、新鲜同种异体神经移植组、新鲜异种神经移植组、化学去细胞同种异体神经移植组和化学去细胞异种神经移植组,每组32只。分别在神经移植术后3、71、4、28 d,每组各取8只小鼠脾脏,淋巴细胞分离液获取淋巴细胞,计数。用荧光标记的CD3+、CD4+、CD8+、CD25+单抗作用后,流式细胞仪检测T淋巴细胞及其亚群百分数,Stata7.0统计软件进行统计处理。结果在7 d1、4 d、28 d各时间点上,化学去细胞神经同种异体和异种神经移植组CD3+、CD4+、CD8+、CD25+阳性T细胞百分率与假手术对照组、自体神经移植组相比无显著差异(P>0.05),而新鲜同种异体及异种神经移植组在相同时间点上的CD3+、CD4+、CD8+、CD25+阳性T细胞百分率与假手术对照组、自体神经移植组相比则有明显增高,差异显著(P<0.05)。结论新鲜同种异体周围神经移植后的排斥属于急性排斥反应。化学去细胞处理的同种异体神经的免疫原性等于或接近于自体神经,要明显低于新鲜同种异体神经移植。

Changes of Splenic T Lymphocytes Subgroups in Mice Grafted with Allogeneic and Heterogenic Peripheral Nerve

Objective To observe the changes of splenic total T lymphocytes and T lymphocyte subgroups during immunologic rejection after allogeneic and heterogenic peripheral nerve grafting so as to delve further into the possibility of the repair of nerve defection.Methods BALB/c mice were divided into sham-operation group,fresh autograft group,fresh allogeneic nerve graft group,fresh heterogenic nerve graft group,chemical de-cell allogeneic nerve graft group and chemical de-cell heterogenic nerve graft group.The different treated nerve grafts were put into the thigh muscle pocket of the mice.The mice were sacrificed respectively on the third,seventh,fourteenth,twenty-eighth postoperative day,and the spleens were sampled.The T lymphocyte counting was carried out and CD3 ,CD4 ,CD8 ,and CD25 cells were measured with the flow cytometer.Results The differences in percentages of CD3 ,CD4 ,CD8 ,and CD25 cells between chemical de-cell allogeneic nerve graft group,chemical de-cell heterogenic nerve graft group and sham-operation group,fresh autograft group on the seventh,fourteenth,twenty-eighth postoperative day were not significant statistically(P>0.05).The percentages of CD3 ,CD4 ,CD8 ,and CD25 cells in fresh allogeneic nerve graft group and fresh heterogenic nerve graft group were much higher than in sham-operation group and fresh autograft group,and the differences were significant statistically(P<0.05).Conclusion The immunogenicities of chemical de-cell allogeneic peripheral nerve and chemical de-cell heterogenic peripheral nerve are very similar to that of self-peripheral nerve,and are much lower than those of fresh allogeneic peripheral nerve and fresh heterogenic peripheral nerve.