新鲜和保存羊膜移植后外周血T细胞变化及免疫病理学研究

目的： 对比研究新鲜羊膜和保存羊膜移植后的免疫反应，客观评价羊膜移植的免疫安全性。方法: 建立BALB/c小鼠皮下埋植实验模型。按不同的埋植物分为单层新鲜羊膜组、双层新鲜羊膜组、甘油保存羊膜组、绒毛膜组（阳性对照）和单纯手术组（阴性对照）；各组又随机分为5个亚组，分别对应术后1周、2周、4周、8周、12周共5个时点。大体观察小鼠的一般情况，流式细胞仪检测外周血CD3+CD25+和CD3+CD71+的表达，免疫组化法定量组织切片中CD3+、CD4+、CD8+的表达。结果: 术后早期，单层新鲜羊膜组和双层新鲜羊膜组的小鼠外周血CD3+CD25+及CD3+CD71+表达，略高于甘油保存羊膜组（P<0.05），但短期内皆可自行回落。各羊膜组移植区组织CD3+、CD4+、CD8+的表达，在术后1周-12周均无显著差异（P>0.05）。各羊膜组的免疫细胞表达均以非特异性为主，显著弱于绒毛膜组（P<0.01）。结论: 新鲜羊膜与保存羊膜在体内表现出几乎无差别的低免疫原性，不会引起由T细胞介导的特异性排斥反应，可视为免疫赦免组织应用。

Changes in peripheral blood T lymphocyte subsets and the immunopathology after fresh or preserved human amniotic membrane transplantation

AIM: To investigate the difference of immunoreaction between fresh and preserved human amniotic membrane(HAM) grafting,and to evaluate the immunologic safety for HAM transplantation.METHODS: Subcutaneous implant models were established in 175 BALB/c mice,which were randomized into five groups with 35 mice each,based on different implants-FAM(fresh amnion),DFAM(double fresh amnion),GPAM(glycerin preserved amnion),chorion(positive control) and merely operation(negative control).Each group was divided into five sub-groups with 7 mice each,according to the observation time: 1 week,2 weeks,4 weeks,8 weeks and 12 weeks after surgery.The expressions of CD3 CD25 and CD3 CD71 in peripheral blood were identified by FACS.The typing of immune cells in the tissues of grafted area was determined by immunohistochemistry.RESULTS: The expressions of CD3 CD25 and CD3 CD71 in fresh AM grafts were more apparent than those in preserved ones(P<0.05),but calmed down initiatively in short time.In all AM groups,only nonspecific immunoreaction as xenografts during early grafting stage was observed,and compared to each other there was no significant difference for the expression of CD3 ,CD4 ,and CD8 (P>0.05).The rejection in chorion groups was much more severe than that in other group(P<0.01) and lasted for a long time.CONCLUSION: Fresh and preserved HAM both seem to be immune-privileged tissue and may be ideal immunocompatibility,which are potentially useful in promoting tissue repair.