摘要
目的研究不负载供者抗原的受者未成熟树突状细胞(immature dendritic cell,imDC)对大鼠肝移植的保护作用,探讨imDC抑制排斥反应的机理。方法以SD大鼠作为供体,雄性Wistar大鼠作为受体,随机分为4组,每组10只,建立肝移植急性排斥反应模型,移植前对受体进行不同的处理。对照组:受体未接受任何处理;环孢素A(CsA)组:术后第2 d开始给予CsA〔10 mg/(kg.d)〕灌胃;成熟树突状细胞(mDC)组:受体术前1 d经阴茎背静脉注射受体来源的mDC 1×106个/只;imDC组:受体术前1 d经阴茎背静脉注射受体来源的imDC1×106个/只。模型建立后10 d各组随机处死5只,取移植肝脏组织行HE染色和免疫组化(FasL/Fas)染色,观察肝脏组织形态结构和排斥反应,根据Banff评分系统判断排斥反应程度;Western blot分析检测各组Foxp3编码的Scurfin蛋白的表达。另留5只大鼠观察术后存活时间;收集血标本,全自动生化分析仪测定ALT、TBIL;双抗体夹心ELISA法检测血清IL-2、IFNγ-、IL-4和IL-10水平。结果CsA组和imDC组术后大鼠中位生存时间明显长于对照组和mDC组,差异有统计学意义(P<0.05)。移植术后各组大鼠血清生化检查:对照组和mDC组ALT及TBIL明显高于CsA组和imDC组(P<0.05);且IL-2和IFNγ-也明显高于CsA组和imDC组(P<0.01),但IL-4和IL-10明显低于CsA组和imDC组(P<0.01)。移植肝脏形态学检测:对照组和mDC组与CsA组和imDC组比较,排斥反应更严重(P<0.05)。Western blot检测:imDC组Foxp3编码的Scurfin蛋白的表达明显高于其他3组(P<0.05)。结论imDC通过阻断间接识别能明显延长移植物存活时间。imDC可能通过以下机理诱导免疫低反应:诱导T细胞凋亡;选择性激活Th2细胞亚群,诱导Th1/Th2偏移;诱导调节性T细胞产生。
Objective To study the mechanism of immune hyporesponsiveness of allograft rejection induced by transfusion nonpufsed allopeptide syngeneic immature dendritic cell （imDC） generated from recipient bone marrow progenitors and to explore a possible strategy for liver allograft protection in clinic. Methods Forty experimental rats were randomly divided into 4 group： control group, cyclosporine A （CsA） group, mature DC （mDC） group and imDC group. In control group, Wistar rats only received liver transplantation. In CsA group, Wistar rats underwent liver transplantation plus CsA treatment [10 mg/（kg · d）]. In mDC group, recipient-derived mDC 1 ×10^6 were infused intravenously through the penile vein to Wistar rats. In imDC group, ImDC with the dose of 1 ×10^6 were injected into Wistar rats via the dorsum vein of penile. In each group, five recipients were killed on the 10th day after transplantation, the other five recipients were left to observe survival time. The levels of ALT, AST, TBIL, IL-2, IFN-γ, IL-4 and IL-10 were detected. The acute rejection and the expression of FasL/Fas in the grafts were detected by HE and immunohistochemieal staining. Western blot was used to detect Scurfin protein expression of CD4＋ CD25＋ T cells. Results The median survival time of the liver allografts in CsA group and imDC group were significantly longer than that in control group and mDC group （P〈0.05）. The levels of ALT and TBIL in control group and mDC group were significantly higher than those in CsA group and imDC group （P〈0. 05）. Compared with CsA group and imDC group, the levels of IL-2 and IFN-γ were higher but the levels of IL-4 and IL-10 were lower in control group and mDC group （P〈0.01）. Slightly or no rejection reaction was found in CsA group and imDC group （P〈0. 05）. The Scurfin protein expressions of CD4＋ CD25＋ T cells of imDC group were significantly higher than those of other three groups. Conclusion Application of nonpufsed allopeptide syngeneic recipient derived
出处
《中国普外基础与临床杂志》
CAS
2009年第1期32-38,共7页
Chinese Journal of Bases and Clinics In General Surgery
基金
云南省科技计划项目(编号:2007CA007)~~
关键词
大鼠
肝移植
排斥反应
未成熟树突状细胞
免疫低反应
Rat
Liver transplantation
Rejection
Immature dendritic cell
Immune hyporesponsiveness
