同种肝移植大鼠转染融合基因人CTLA4-Ig抑制急性排斥反应  

Gene transfer of huCTLA4-Ig to inhibit the acute rejection of liver allograft in rats

作　　者：朱斌[1] 杨甲梅[1] 钱其军[1] 王孟龙[1] 崔贞福[1] 李玲芳[1] 吴孟超[1] 

机构地区：[1]第二军医大学附属东方肝胆外科医院肝胆外科研究所,上海200438

出　　处：《中华器官移植杂志》2005年第4期199-202,共4页Chinese Journal of Organ Transplantation

基　　金：国家自然科学基金资助项目(39900138)上海市科学技术发展基金资助项目(004119014)

摘　　要：目的观察同种肝移植大鼠转染融合基因人CTLA4-Ig(huCTLA4-Ig)对急性排斥反应的抑制作用。方法供者为BN大鼠,受者为Lew大鼠。将受者分为三组,A组(5只):为空白对照组,供肝冷保存时不感染腺病毒;B组(4只):为阴性对照组,供肝冷保存时感染携带报告基因GFP 的重组腺病毒;C组(5只):为基因治疗组,供肝冷保存时,先体外后体内感染携带融合基因huCT- LA4-Ig的重组腺病毒;观察该治疗基因在受者体内的表达及其对急性排斥反应的抑制作用。结果(1)A组与B组受者均在3周内死亡,A、B组平均生存时间分别为(16.6±2.5)d和(15.5±3.1)d,C 组受者均能获得长期生存(>150 d);C组与A、B组生存期比较,差异有统计学意义(P<0.01)。(2) 移植肝病理学检查发现:术后8 d,A组与B组移植肝均发生严重的细胞性排斥。C组术后8 d,可见移植肝轻、中度的汇管区炎症,但组织损伤程度显著减轻;术后150d,可发现单核细胞浸润,但胆管保存完好,无血管损伤证据。(3)术前大鼠血清白细胞介素2(IL-2)浓度为(362.09±45.84)ng/L;肝移植术后,A、B组血清IL-2的浓度即升到较高水平(约为术前的1.5-2.5倍),而C组却始终维持在一个较低水平(接近或低于术前水平);术后3、7 d,C组血清IL-2的浓度与A、B组比较,差异均有统计学意义(P<0.01)。Objective To observe the effect of gene transfer of huCTLA4-Ig to inhibit the acute rejection of liver allograft in rats. Methods With AdEasy vector system, the recombinant adenovirus containing huCTLA4-Ig gene was constructed. Using ex vivo gene transfer technique, exogenous gene was introduced to the liver graft during cold preservation and expressed locally in the graft. The effect of inhibition of acute rejection and inducing liver graft tolerance was observed. Results No recipients in group A (without any treatment, n = 5) or group B (treated with Ad-GFP, n - 4) died within 3 weeks after transplantation and severe acute rejection (massive periportal infiltration, endothelilitis, damage to biliary epithelium and severe tissue destruction) was confirmed pathologically in the graft. In contrast, all recipients in group C (treated with Ad-huCTLA4-Ig, n- 5) achieved long-term liver allograft survival (>150 days). Histological examination of Ad-huCTLA4-Ig transduced allografts demonstrated a mild to moderate periportal inflammation and mild injury to liver graft on day 8 postt-ransplant. A mild mononuclear infiltration was observed; however, there was complete preservation of the bile ducts and no evidence of vascular injury on day 150 posttransplant. The mean IL-2 concentration in serum was (362. 09±45. 84)ng/L on day 1 pretransplant. In control animals (groups A and B), serum IL-2 concentration was elevated to a high level within 7 days posttransplant, which was a-bout 1.5 to 2. 5 times as much as that before transplant. In contrast, in huCTLA4-Ig-treated animals (group C), IL-2 concentration in serum was maintained at a relative low level, which was near or less than that before transplant (P<0. 01). Conclusions Using ex vivo gene transfer technique, huCTLA4-Ig gene can be introduced to the liver graft during cold preservation. The modified graft can express and excrete immunoregulatory protein locally, which can suppress acute alloimmune response and is re sponsible for prolongation of graft survival without

关 键 词：急性排斥反应 同种肝移植 融合基因 CTLA4-IG 大鼠 转染 血清IL-2 重组腺病毒 血清白细胞介素 同种异体移植肝 单核细胞浸润 基因转移系统 抑制作用 病理学检查 细胞性排斥 肝移植术后 分泌性表达 免疫抑制剂 冷保存 报告基因 

分 类 号：R657.3[医药卫生—外科学]