【摘要】： 目的:探討重組腺病毒載體(AdOX40Ig)介導的OX40-Ig局部基因治療在阻斷OX40/OX40L共刺激途徑抑制復合組織同種異體移植急性排斥反應中的作用。 方法:構建了含人OX40胞外段和人IgG1Fc段的重組腺病毒表達載體AdOX40Ig。應用一種新的體外基因轉染方法,使移植物局部能夠分泌免疫調節(jié)分子OX40-Ig融合蛋白。采用近交系大鼠下腹壁淺動脈(superficial inferior epigastric artery,SIEA)皮瓣移植模型,檢驗AdOX40Ig介導的OX40-Ig局部表達對阻斷OX40/OX40L共刺激途徑延長移植物存活時間的作用。實驗分為5組,分別比較了單純地移植術前AdOX40Ig灌注轉染治療方案、小劑量雷帕霉素治療方案及二者相結合對移植物存活時間的影響。應用組織病理學分級對移植物存活進行評估。 結果:移植物經體外基因轉染后,自身能夠高表達OX40-Ig蛋白。術后第7天未處理組(A組,7.8±1.2d)和AdEGFP轉染組(B組,7.1±1.2d)的皮瓣均發(fā)生了GradeⅢ級免疫排斥反應。單獨使用AdOX40Ig灌注轉染組(C組,8.3±0.8d)的皮瓣存活時間與A及B組比較并無顯著差異(p0.05);然而,聯合小劑量雷帕霉素治療后,皮瓣的平均存活時間可達18.6±1.3d,與單獨應用小劑量雷帕霉素組相比有顯著差異(E組,13.5±0.5d,p0.01)。 結論:研究表明,OX40-Ig局部的免疫調節(jié)作用與小劑量的雷帕霉素相協同可延長移植物的存活時間。該方法有效地減小了術后早期全身性免疫抑制劑的用量,為異體復合組織移植免疫耐受的誘導提供了新思路。
[Abstract]:Aim: to investigate the role of recombinant adenovirus vector (AdOX40Ig)-mediated OX40-Ig gene therapy in the suppression of acute allograft rejection by blocking the OX40/OX40L costimulatory pathway. Methods: recombinant adenovirus expression vector AdOX40Ig. containing extracellular and IgG1Fc segments of human OX40 was constructed. A novel in vitro gene transfection method was developed to secrete OX40-Ig fusion protein. The graft model of (superficial inferior epigastric artery,SIEA flap of inferior epigastric artery in inbred rat was used to examine the effect of AdOX40Ig mediated local expression of OX40-Ig on prolonging the survival time of graft by blocking OX40/OX40L costimulatory pathway. The experiment was divided into 5 groups. The effects of AdOX40Ig perfusion transfection regimen, low dose rapamycin regimen and their combination on graft survival time were compared before transplantation alone. Histopathological grading was used to evaluate graft survival. Results: after gene transfection in vitro, the graft could overexpress OX40-Ig protein. On the 7th day after operation, Grade grade 鈪，

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