【摘要】： 目的:探討重組腺病毒載體(AdOX40Ig)介導(dǎo)的OX40-Ig局部基因治療在阻斷OX40/OX40L共刺激途徑抑制復(fù)合組織同種異體移植急性排斥反應(yīng)中的作用。 方法:構(gòu)建了含人OX40胞外段和人IgG1Fc段的重組腺病毒表達(dá)載體AdOX40Ig。應(yīng)用一種新的體外基因轉(zhuǎn)染方法,使移植物局部能夠分泌免疫調(diào)節(jié)分子OX40-Ig融合蛋白。采用近交系大鼠下腹壁淺動(dòng)脈(superficial inferior epigastric artery,SIEA)皮瓣移植模型,檢驗(yàn)AdOX40Ig介導(dǎo)的OX40-Ig局部表達(dá)對(duì)阻斷OX40/OX40L共刺激途徑延長(zhǎng)移植物存活時(shí)間的作用。實(shí)驗(yàn)分為5組,分別比較了單純地移植術(shù)前AdOX40Ig灌注轉(zhuǎn)染治療方案、小劑量雷帕霉素治療方案及二者相結(jié)合對(duì)移植物存活時(shí)間的影響。應(yīng)用組織病理學(xué)分級(jí)對(duì)移植物存活進(jìn)行評(píng)估。 結(jié)果:移植物經(jīng)體外基因轉(zhuǎn)染后,自身能夠高表達(dá)OX40-Ig蛋白。術(shù)后第7天未處理組(A組,7.8±1.2d)和AdEGFP轉(zhuǎn)染組(B組,7.1±1.2d)的皮瓣均發(fā)生了GradeⅢ級(jí)免疫排斥反應(yīng)。單獨(dú)使用AdOX40Ig灌注轉(zhuǎn)染組(C組,8.3±0.8d)的皮瓣存活時(shí)間與A及B組比較并無(wú)顯著差異(p0.05);然而,聯(lián)合小劑量雷帕霉素治療后,皮瓣的平均存活時(shí)間可達(dá)18.6±1.3d,與單獨(dú)應(yīng)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(jié)論:研究表明,OX40-Ig局部的免疫調(diào)節(jié)作用與小劑量的雷帕霉素相協(xié)同可延長(zhǎng)移植物的存活時(shí)間。該方法有效地減小了術(shù)后早期全身性免疫抑制劑的用量,為異體復(fù)合組織移植免疫耐受的誘導(dǎo)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，

本文編號(hào)：2205564