移植物局部转染CTLA4Ig基因对大鼠小肠移植排斥反应的预防作用  

Treatment of CTLA4Ig gene for the rejection of small bowel allografts in rats

作　　者：王一芳[1] 赖福生[1] 许爱刚[1] 吴文溪[1] 

机构地区：[1]南京医科大学第一附属医院胃肠外科,210029

出　　处：《中华器官移植杂志》2005年第1期33-35,共3页Chinese Journal of Organ Transplantation

基　　金：江苏省教育厅重点学科基金资助项目(H200212)

摘　　要：目的探讨CTLA4Ig基因在移植小肠局部转染表达及表达产物对排斥反应的预防作用。方法大鼠小肠移植前,经肠系膜上动脉给供肠灌注脂质体包裹的CTLA4IgcDNA重组质粒,术后应用组织免疫学及逆转录聚合酶链反应(RTPCR)检测移植小肠中CTLA4Ig转基因产物的表达,并行组织病理学检查,同时设立无CTLA4Ig基因转染的对照组。结果对照组(n=8)的移植小肠在术后7～14d全部坏死;实验组(n=18)7只受鼠的移植小肠于术后56～90d坏死,11只受鼠的移植小肠于术后90d时仍存活良好。对照组移植物组织切片见单核细胞广泛浸润,肠壁肌层及黏膜出血坏死,绒毛及隐窝结构消失;实验组存活超过90d者组织切片仅见少量单核细胞浸润,黏膜轻度增厚,但无明显的肠绒毛缩短及隐窝细胞减少等改变。组织免疫学及RTPCR结果显示,在移植后28d内移植小肠中有CTLA4Ig转基因产物的表达,但第56d及90d则检测不到其表达。结论经肠系膜上动脉体外灌注脂质体包裹的CTLA4IgcDNA重组质粒可有效转染移植小肠,CTLA4Ig基因在移植小肠局部转染对术后的排斥反应具有预防作用。Objective To evaluate the local expression of CTLA4Ig gene in small intestines and its action on preventing rejection of the small bowel allografts. Methods The donor small bowels were perfused ex vivo with CTLA4Ig cDNA reconstructed plasmid packaged with lipofectin vector via intra-superior mesenteric artery before transplantation. After transplantation, the CTLA4Ig transgene expression in the small bowel allografts was assessed by immunohistology and reverse transcriptase polymerase chain reaction (RT-PCR), and the histological examination of the allografts was performed. The donor small bowels not receiving CTLA4Ig cDNA transfection served as the control group. Results Necrosis of implanted small bowel was found 7-14 days after transplantation in all animals of the control group (n=8). In gene transfer group (n=18), necrosis was found in 7 allografts 56-90 days after transplantation, and 11 cases of implanted small bowels survived more than 90 days in the recipients. For histological examination, the intestinal allografts in control group showed infiltration of extensive mononuclear cells, and the villi and crypts were completely destructed, with hemorrhage and necrosis in muscularis and mucosa. However, the small bowel allografts with survival time being more than 90 days in gene transfer group showed only a few mononuclear cells infiltration, slight mucosa ~incrassation , no significant villi shortening and crypt cells reduction. The CTLA4Ig expression in the ~allografts in gene transfer group was detected by immunohistology and RT-PCR within 28 days after transplantation, but no expression of CTLA4Ig transgene was detectable at 56th day and 90th day.Conclusion A single ex vivo intra-superior mesenteric artery infusion of CTLA4Ig cDNA ~recons -tructed plasmid packaged with lipofectin can induce efficient transduction of the small intestine. ~Trans -fection of CTLA4Ig gene in small bowel allografts can prevent rejection after transplantation.

关 键 词：移植物 转染 CTLA4IG基因 大鼠 小肠 移植排斥反应 预防作用 

分 类 号：R656.7[医药卫生—外科学]