抗人C5单克隆抗体对补体介导的超急性排斥反应的抑制作用  

Inhibitory effects of anti-human C5 monoclonal antibody on complement-mediated hyperacute rejection in vitro

作　　者：董秀华[1] 付小锁[4] 卿恩明[1] 赵莉敏[2] 刘晓明[1] 任发成[1] 卢家凯[1] 王学勇[1] 孟旭[3] 

机构地区：[1]首都医科大学附属北京安贞医院麻醉科 ,100029 [2]首都医科大学附属北京安贞医院实验研究中心 ,100029 [3]首都医科大学附属北京安贞医院 心脏外科,100029 [4]首都医科大学基础医学院生理学系

出　　处：《中华器官移植杂志》2009年第4期215-217,共3页Chinese Journal of Organ Transplantation

基　　金：首都医科大学基础-临床合作基金（2006JL43）

摘　　要：目的研究抗人C5单克隆抗体对补体介导的超急性排斥反应的抑制作用。方法获取8例临床心脏移植供者的主动脉血20ml和主动脉血管，分别用于提取淋巴细胞和主动脉内皮细胞。同时，采集相应8例心脏移植受者移植前的中心静脉血20ml和外周静脉血5ml，分别用于提取淋巴细胞和制备补体。供者主动脉内皮细胞经培养后，依次加入供、受者淋巴细胞混合培养上清50μl和受者静脉血浆（含补体）20μl，建立主动脉内皮细胞超急性排斥反应模型。模型建立后，将供者主动脉内皮细胞平均分为2组，C5组：加入20mg／L的抗人C5单克隆抗体20μl；对照组：加入M200培养液20μl。经台盼蓝染色，倒置显微镜下观察主动脉内皮细胞的染色情况及活性，并测定主动脉内皮细胞刺激指数（SI）。结果经观察，对照组大部分主动脉内皮细胞蓝染、肿胀死亡，而C5组大部分主动脉内皮细胞未见蓝染，存活良好；对照组和C5组吸光度值分别为1．528±0．046和1．644±0．058，两组比较，差异有统计学意义（P〈0．001），SI为1．076±0．038。结论在离体主动脉内皮细胞超急性排斥反应模型中，抗人C5单克隆抗体可以抑制补体介导的主动脉内皮细胞溶解。Objective To observe the inhibitory effect of anti-human C5 monoclonal antibody on the complement-mediated hyperacute rejection. Methods The blood and the aortic vessels were obtained from 8 heart donors respectively to separate lymphoeytes and culture the aortic endothelial cells of donors respectively. The blood from 8 heart-recipients was also drawn to separate the lymphocytes and plasma which contained the complement elements. The hyperacute rejection model of aortic endothelial cells was established when the cultured donor aortic endothelial cells were added with the supernatant of mixed one-way lymphocyte culture and recipient＇ s plasma which contained complement elements. The models were divided into 2 groups： C5 group receiving 20 mg/L antihuman C5 monoclonal antibody （20μl）, and placebo group treated with equal volume of M200 culture medium. The aortic endothelial cell viability was observed by trypan blue staining and the stimulation index （SI） was determined by MTT assay. Results The trypan blue staining showed that most aortic endothelial cells in placebo group were blue dyed, swollen and died, while the donor aortic endothelial cells in C5 group were alive without dying blue. The absorbance （A） values had significant difference between placebo group （A value was 1. 528 ± 0. 046） and C5 group （A value was 1. 644 ± 0. 058） （P〈0. 001）. The SI was 1. 076 ± 0. 038. Conclusion Anti-human C5 monoclonal antibody can inhibit the complement-mediated lysis of donor aortic endothelial cells in the hyperacute rejection model of aortic endothelial cells in vitro.

关 键 词：心脏移植 移植物排斥 补体C5 

分 类 号：R686[医药卫生—骨科学]