供肝局部转染人细胞毒性T淋巴细胞相关抗原4免疫球蛋白的重组腺相关病毒诱导大鼠同种肝移植免疫耐受  

Induced liver allograft immune tolerance in rats by locally transferred rAAV-hCTLA4Ig

作　　者：苏志雷[1] 韩德恩[1] 姜明山[1] 邰升[1] 

机构地区：[1]哈尔滨医科大学附属第二医院肝脏外科,150086

出　　处：《中华实验外科杂志》2007年第9期1065-1066,共2页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(304004160);哈尔滨市科技攻关计划项目(2004AA9CS196-2)

摘　　要：目的利用人细胞毒性T淋巴细胞相关抗原4免疫球蛋白的重组腺相关病毒(rAAV-hCTLA4Ig)局部基因转染供肝诱导大鼠同种肝移植免疫耐受。方法供体为DA大鼠,受体为LEW大鼠,分为以下4组:(A)同基因对照组(LEW-LEW);(B)生理盐水对照组;(c)rAAV- EGFP(增强型绿色荧光蛋白)对照组;(D)rAAV-hCTLA4Ig灌注组。大鼠原位肝移植前6周,经门静脉灌注采用血管夹闭法对供肝进行基因转染。结果D组大鼠平均生存期显著高于C组(10.8±1.0)d及B组(11.6±1.1)d,B组与D组间,C组与D组间,生存期差异有统计学意义(P<0.01)。D组血浆及移植肝中可以持续检测到hCTLA4Ig的表达。术后1周移植肝病理检查显示B组、C组均有严重免疫排斥反应,免疫组织化学显示大量CD4^+、CD8^+T淋巴细胞浸润;而D组仅有轻、中度炎症反应,少量CD4^+、CD8^+T淋巴细胞浸润。结论移植术前6周对供肝体内经门静脉灌注采用血管夹闭技术基因转染rAAV-hCTLA4Ig可以诱导大鼠同种肝移植免疫耐受。Objective To induce liver allograft immune tolerance in rats by locally transferred rAAV-hCTLA4Ig （ recombined Adeno associated virus- human cytotoxic T lymphocyte associated antigen 4 immunoglobulin）. Methods Dark Agouti and Lewis rats were used as donors and recipients, respectively, in four experimental groups ： （A） syngeneic control, （B） no treatment, （C） rAAV-EGFP control, （D） rAAV-hCTLA4Ig. The transfection of all genes was accomplished 6 weeks before the transplantation by portal vein infusion using a vascular clamping technique. Results The mean survival time （ 〉 100 days） in group D was significantly longer than in group C （ 10.8 ± 1.0） days and group B （ 11.6 ± 1.1 ） days, and there was significant difference between group B and D, and between group C and D （P 〈 0.01 ）. In group D,the expression of hCTLA4Ig in serum and liver grafts was detected continuously. One week after operation, there was severe rejection responses and a large number of CD4^＋ and CD8^＋ T lymphocytes infiltrated in groups B and C, while in groups D, there was only mild or moderate inflammatory reaction and few T lymphocytes infiltration. Conclusion An immune tolerance liver graft could be achieved by gene transfer of rAAV-hCTLA4Ig through portal vein infusion using a clamping technique.

关 键 词：肝移植 腺相关病毒 免疫耐受 

分 类 号：R657.3[医药卫生—外科学] R392.4[医药卫生—临床医学]