恒河猴肝移植模型急性排斥反应时T细胞亚群的变化（英文）

Variation of T cell subset during acute rejection after liver transplantation in rhesus monkeys

背景:寻找肝移植急性排斥反应的早期诊断指标,可快速而有效的评估移植后排斥反应的风险,而T淋巴细胞在急性排斥反应中发挥主要作用。目的:观察恒河猴肝移植后血液T淋巴细胞亚群的变化与急性排斥反应的关系。方法:采用改良血管袖套+胆道支撑管+动脉吻合法建立16例稳定的恒河猴肝移植模型,并将模型随机分为2组:实验组围手术期不给予免疫抑制治疗,对照组围手术期给予免疫抑制治疗,分别在移植后6,12,24,72 h共4个时间点收集血标本和肝组织,全自动生化分析仪测定丙氨酸氨基转移酶、天冬氨酸氨基转移酶、总胆红素,用流式细胞仪检测血CD4+/CD8+水平,然后取移植后肝脏组织进行苏木精-伊红染色以观察肝脏组织形态结构的改变,并根据Banff评分系统判断排斥反应程度。结果与结论:实验组恒河猴肝移植后12,24,72 h均有急性排斥反应发生,其中实验组丙氨酸氨基转移酶、天冬氨酸氨基转移酶和总胆红素在移植后24,72 h表达水平明显高于对照组(P<0.05),实验组和对照组CD4+/CD8+的表达在移植后6 h即开始增高,实验组增高最明显,实验组移植后24,72 h CD4+/CD8+的表达明显高于对照组(P<0.05),实验组移植后72 h肝脏组织形态病理改变程度重于对照组,Banff评分高于对照组(P<0.05)。说明在肝移植后急性排斥反应早期,CD4+/CD8+的表达变化早于肝组织病理学及肝功能的改变。肝移植后血CD4+/CD8+水平的升高提示人体细胞免疫功能增强,对肝移植后急性排斥反应的早期诊断具重要意义。

Abstract BACKGROUND： Looking for the early diagnosis of acute rejection indicators after liver transplantation can assess the risk after liver transplantation quickly and effectively, and T lymphocytes play the significant role in acute rejection. OBJECTIVE：To observe the relationship between acute rejection and variation of expression of T cel subset in blood after liver transplantation in rhesus monkey. METHODS： The sixteen liver transplant models in rhesus monkey which were constructed successfuly by the method of ＂double-cuff and one support tube＂ were divided into two groups randomly： experiment group （no treated by immunosuppressant in perioperative period） and control group （treated by immunosuppressant in perioperative period）. Then the blood specimen and liver tissue respectively were colected at 6, 12, 24 and 72 hours after operation. The levels of alanine transferase, aspartate aminotransferase, and total bilirubin were detected with the fuly automatic biochemical analyser. The levels of CD4＋/CD8＋were tested by flow cytometry. The liver tissue in rhesus monkey after liver transplantation was detected by hematoxylin-eosin staining. The degree of acute rejection was evaluated by Banff Score System. RESULTS AND CONCLUSION： Acute rejection appeared in the experiment group at 12, 24, and 72 hours after liver transplantation. Levels of alanine transferase, aspartate aminotransferase, and total bilirubin were significantly higher in the experimental group than in the control group at 24 and 72 hours after transplantation （P 〈 0.05）. The expression of CD4＋/CD8＋of the experiment group and control group began to rise at 6 hours after surgery, but the experiment group increased the most obvious. CD4＋/CD8＋ expression was significantly greater in the experimental group than in the control group at 24 and 72 hours after transplantation （P 〈0.05）. Morphological pathology was severer, and Banff score was higher in the experiment group than in the control group at 72 hours （P 〈0.05）. These data suggested that the variation of expression of CD4＋/CD8＋was earlier than the change of liver tissue pathology and the change of liver function in the early acute rejection after liver transplantation. The rise of level of CD4＋/CD8＋ after liver transplantation indicated the increase of celular immunity in body, which had an important role in the early diagnosis of acute rejection after liver transplantation.