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目的 探讨CTLA 4Ig基因重组腺病毒 (AdvCTLA 4Ig)有效转染大鼠肾脏的方法并初步观察该基因转染对移植肾存活的影响。方法 AdvCTLA 4Ig对大鼠肾脏分别转染 3h ,包括 :4℃下转染 (A组 )、经肾动脉注射在体转染 (B组 )、阻断肝脏血流以及肾血管以下躯体血流后经肾动脉注射在体转染 (C组 )以及对照 ;以免疫荧光法检测基因转染效率 ;以转染效率最高组肾脏行同种移植 ,未转染组为对照 ,观察移植肾存活时间。结果 A组、B组以及对照组肾脏内未见荧光显示 ,C组肾小管显示阳性荧光 ,肾小球内未见荧光显示 ;各组肾脏光镜下结构正常 ;转染组移植肾存活时间较对照组明显延长 ( 2 6 6± 6 9)dvs ( 8 1± 1 60 )d ,P <0 0 1。结论 高滴度CTLA 4Ig基因重组腺病毒可使在体肾脏中肾小管获得有效转染 ;该基因转染的肾脏移植后存活时间明显延长
Objective To investigate the method of transfection of rat CTL 4Ig recombinant adenovirus (AdvCTLA 4Ig) into rat kidney and to investigate the effect of transfection on the survival of renal allografts. Methods AdvCTLA 4Ig was transfected into rat kidneys for 3h, including transfection at 4 ℃ (group A), transfection by renal artery in group B (group B), blocking hepatic blood flow and blood flow below the renal vessels Transplantation was performed in renal artery (group C) and control group. The efficiency of gene transfection was detected by immunofluorescence. Allograft and non-transfected group were used as control group to observe the survival time of transplanted kidney. Results No fluorescence was observed in the kidneys of group A, group B and control group. The renal tubules of group C showed positive fluorescence and no glomerular fluorescence was observed. The structure of kidney in each group was normal under light microscope. The control group was significantly prolonged (266 ± 69) dvs (81 ± 160) d, P <0.01. Conclusion High titer CTLA 4Ig recombinant adenovirus can effectively translocate renal tubules in vivo and the survival time of transplanted kidney after transplanting is significantly longer