抗白细胞介素6单克隆抗体治疗延长小鼠同种心脏移植物存活时间的实验研究  

Prolongation of Murine Cardiac Allograft Survival Mediated by Anti-IL-6 Monoclonal Antibody Therapy

作　　者：雷钧[1] 何凡[1] 吴敏[1] 郑翔[1] 李舒媛[1] 陈知水[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院器官移植研究所,器官移植教育部重点实验章,器官移植卫生部重点实验室,武汉430030

出　　处：《华中科技大学学报（医学版）》2010年第2期197-201,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：湖北省科技攻关重大项目资助(No.2006AA301A06)

摘　　要：目的观察应用抗白细胞介素6(IL-6)单克隆抗体(anti-IL-6 monoclonal antibody,anti-IL-6 mAb)对同种异体小鼠心脏移植排斥反应的抑制作用,并探讨其可能的作用机制。方法建立小鼠颈部异位心脏移植模型。实验动物随机分为同系移植组、移植治疗组(每日注射3 g/L anti-IL-6 mAb 0.1 mL)和移植模型组(注射等量生理盐水)。ELISA法检测受鼠移植术后第3、6天外周血IL-6的表达水平,观察各组移植心脏存活时间,病理分析检测排斥反应程度,流式细胞术(FCM)检测各组受鼠脾脏及移植心脏中CD4+CD25+调节性T细胞比例变化,RT-PCR检测移植心脏IL-6I、FN-γI、L-17、RORγt和Foxp3 mRNA的表达水平。结果①治疗组外周血IL-6表达水平较模型组明显降低(P<0.01);②治疗组移植心脏存活时间较模型组明显延长[(22.0±2.3)d vs(7.7±0.8)d,P<0.01],排斥反应病理分级明显下降(Stanford 2级);③治疗组移植心脏中IFN-γI、L-6I、L-17和RORγt mRNA表达水平较模型组显著下降(均P<0.05),脾脏及移植心脏中浸润的CD4+CD25+调节性T细胞比例及Foxp3 mRNA表达水平均明显升高,与模型组比较差异有统计学意义(均P<0.05)。结论anti-IL-6 mAb治疗可抑制Th1、Th17型细胞因子的分泌,促进CD4+CD25+Foxp3+T细胞产生,抑制单个核细胞浸润并有效延长小鼠同种移植心脏的存活时间。Objective To investigate the efficacy of anti-IL-6 monoclonal antibody（anti-IL-6 mAb） in delaying acute allograft rejection in a mouse model of heart transplantation and analyze the biological mechanisms underlying the protective effects.Methods The heterotopic cardiac transplantation models in mice were established.The mice were randomly divided into three groups：the isograft transplantation group,the transplantation model group and the transplantation treatment group.The animals in the treatment group were treated daily with anti-IL-6 mAb（0.1 mL,3 g/L）,and those in the model group received the same volume of normal saline.The serum levels of IL-6 were measured by using enzyme-linked immunosorbent assay（ELISA） at the 3rd and 6th day after transplantation.The histopathological changes and the allograft mean survival time were also observed.The mRNA expression of Foxp3 and pro-inflammatory cytokine in recipient graft was detected by using RT-PCR.Results In the animals treated with anti-IL-6 mAb cardiac allograft survival was significantly prolonged as compared with those treated with normal saline（22.0±2.3 days vs 7.7±0.8 days,P0.01）.IL-6 levels in serum were significantly reduced after anti-IL-6 mAb administration.In the hearts treated with anti-IL-6 mAb,the levels of IFN-γ,IL-6,IL-17 and RORγt were significantly reduced,whereas Foxp3 levels were increased.Heart allografts pre-treated with anti-IL-6 mAb had a significant higher proportion of Tregs than those pre-treated with normal saline（P0.05）.Conclusion Anti-IL-6 mAb overcomes the activation of effector T cells and promotes the induction of regulatory T cells for acute rejection in mouse cardiac allografts,thereby mediating prolonged graft survival.Intragraft IL-6 inhibition may be useful as an adjuvant therapy to systemic immunosuppression in heart transplantation.

关 键 词：心脏移植 急性排斥反应 白细胞介素6 单克隆抗体 

分 类 号：R654.28[医药卫生—外科学]