重组腺病毒介导的OX40-Ig局部基因治疗对大鼠同种异体复合组织移植的免疫调节

发布时间：2018-08-26 16:57

【摘要】： 目的:探讨重组腺病毒载体(AdOX40Ig)介导的OX40-Ig局部基因治疗在阻断OX40/OX40L共刺激途径抑制复合组织同种异体移植急性排斥反应中的作用。 方法:构建了含人OX40胞外段和人IgG1Fc段的重组腺病毒表达载体AdOX40Ig。应用一种新的体外基因转染方法,使移植物局部能够分泌免疫调节分子OX40-Ig融合蛋白。采用近交系大鼠下腹壁浅动脉(superficial inferior epigastric artery,SIEA)皮瓣移植模型,检验AdOX40Ig介导的OX40-Ig局部表达对阻断OX40/OX40L共刺激途径延长移植物存活时间的作用。实验分为5组,分别比较了单纯地移植术前AdOX40Ig灌注转染治疗方案、小剂量雷帕霉素治疗方案及二者相结合对移植物存活时间的影响。应用组织病理学分级对移植物存活进行评估。 结果:移植物经体外基因转染后,自身能够高表达OX40-Ig蛋白。术后第7天未处理组(A组,7.8±1.2d)和AdEGFP转染组(B组,7.1±1.2d)的皮瓣均发生了GradeⅢ级免疫排斥反应。单独使用AdOX40Ig灌注转染组(C组,8.3±0.8d)的皮瓣存活时间与A及B组比较并无显著差异(p0.05);然而,联合小剂量雷帕霉素治疗后,皮瓣的平均存活时间可达18.6±1.3d,与单独应用小剂量雷帕霉素组相比有显著差异(E组,13.5±0.5d,p0.01)。 结论:研究表明,OX40-Ig局部的免疫调节作用与小剂量的雷帕霉素相协同可延长移植物的存活时间。该方法有效地减小了术后早期全身性免疫抑制剂的用量,为异体复合组织移植免疫耐受的诱导提供了新思路。
[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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