Analysis of rejection mechanism, aiming for the establishment of novel immunosuppression in small bowel transplantation

背景：临床活体小肠移植的经验促使我们对以下问题进行研究，这些问题一直是制约移植物和/或患者存活的障碍。早期和明确的排斥诊断更复杂的排斥控制方法猪SBT模型：由于人和猪在解剖学、生理学和免疫学上的相似性，本实验采用猪作为实验动物。采用供受者混合淋巴细胞反应建立一期同种异体供、受者SBT模型。标本取材：(1)通过中线给药和采血；(2)纤维支气管镜(使用临床儿童纤维)和活检；(2)建立猪原位SBT模型，安全、规范地实施这些技术，为临床移植在同一水平监测排斥反应过程。结果：根据宏观和显微镜观察结果，REJ-…的严重程度本文分析了临床SBT的关键药物他克莫司免疫抑制作用下移植肾细胞凋亡小体计数的变化及其与细胞凋亡的关系。发生排斥反应的移植物标本的凋亡小体计数(ABC)(6.59/10个)明显高于非排斥移植物(0.773个/10个)，差异有统计学意义(P<0.05)。排斥反应的严重程度与ABC无关。免疫组织化学研究分析了细胞凋亡的诱导途径。在肉眼和/或组织学排斥反应的同时或之前，观察到隐窝周围的CD8+淋巴细胞和FasL+淋巴细胞的浸润以及上皮细胞上肿瘤坏死因子受体的表达增加。未来的展望和计划：现在可以利用上述指标更详细地分析猪SBT的排斥反应。利用这些指标，我们正在进行进一步的研究项目，如局部应用免疫抑制剂或诱导耐受。这些指标将应用于我们即将到来的诊断排斥反应的临床SBT病例。较少

Background : The experience with clinical living donor SBT(small bowel transplantation) motivate us to research on the following problems, which have been the obstacles to the improvement of graft and/or patient survival.Earlier and definite diagnosis of rejectionMore sophisticated way to control rejectionPig SBT model : Pigs are adopted for our experiment because of the anatomical, physiological and immunological similarity between humans and pigs. We established one-staged orthotopic SBT model using donors and recipients which showed alloreactivity with mixed lymphocyte reaction.Sampling of specimens :a) drug administration and blood sampling through central lineb) fiberscopy (using clinical pediatric fiber) and biopsyWe established techniques above with pig SBT model and are performing these safely and regularly, which enable us to monitor the rejection process at the same level for clinical transplantation.Results : Based on the macroscopic and microscopic findings, severity of rej … More ection and its relationship to ABC (apoptotic body counts) were analyzed under the immunosuppression with Tacrolimus, which has been the key drug for clinical SBT.Samples from grafts with rejection presented higher ABC (6.59/l0crypts) than those from non-rejected grafts (ABC ; 0.773/l0crypts), which showed statistical difference. There was no correlation between the severity of rejection and ABC.Immunohistochemical studies were performed to analyze the induction pathway to apoptosis. Infiltration of CD8+ lymphocytes and FasL+ lymphocytes around crypts and increased expression of TNF receptor on epithelial cells were observed at the same time with, or prior to, macroscopic and/or histological rejection.Future prospects and plans : The rejection in pig SBT now can be analyzed more in detail with the indicators above. Using the indicators, we are moving on to the further research projects such as local administration of immunosuppressants or tolerance induction. These indicators will be applied for our coming clinical SBT cases for the diagnosis of rejection. Less