转染细胞毒性T淋巴细胞相关抗原4免疫球蛋白基因的同供体大鼠树突状细胞对胰岛移植物存活的影响  

Effect cytotoxic T lymphocyte associated antigen-4 immunoglobulin gene transfer to the same donor dendritic cells on rat pancreatic islet allografts

作　　者：汤志刚[1] 王春友[2] 胡何节[1] 

机构地区：[1]安徽省立医院普通外科,合肥230001 [2]华中科技大学同济医学院协和医院胰腺外科治疗中心

出　　处：《中华实验外科杂志》2005年第4期407-409,共3页Chinese Journal of Experimental Surgery

摘　　要：目的　探讨转染细胞毒性T淋巴细胞相关抗原4(转染CTLA4Ig基因)的同供体大鼠DC细胞对同种大鼠胰岛移植的影响。方法　链尿菌素(STZ) 60mg/kg体重腹腔内注射制作SD大鼠糖尿病模型,胶原酶法分离、Ficoll 40 0密度梯度离心法纯化胰岛,GM CSF +IL 4诱生培育的方法,获得高纯度的DC ,含目的基因CTLA4Ig重组腺病毒AdvCTLA4Ig ,转染同供体DC细胞,与胰岛细胞同时移植于糖尿病受体大鼠肾包膜下,免疫组织化学、Dot ELISA、逆转录 聚合酶链反应(RT PCR)检测CTLA4Ig基因在实验组和对照组DC细胞中的表达,并检测受体大鼠的血糖浓度变化,同时观察受体存活情况。结果　实验组DC细胞有CTLA4Ig表达,而对照组DC细胞没有CTLA4Ig表达,实验组正常血糖维持时间(17.3±2 .4)d较对照组(10 .1±1.5 )d、空白对照组(8.3±1.2 )d显著延长,实验组、对照组和空白对照组受体大鼠存活天数分别为(3 4.5±3 .4)d、(14 .7±2 .3 )d和(11.2±1.4)d ,实验组高于对照组(P <0 .0 1)。结论　表达CTLA4Ig基因的DC细胞可能诱异胰岛移植免疫耐受。Objective To investigate the effect of cytotoxic T lymphocyte associated antigen-4 immunoglobulin (CTLA4Ig) gene transfer to the same donor dendritic cells (DC) on rat pancreatic islet allografts.Methods The SD rat diabetic model was developed by intraperitoneal injection of streptozotocin 60 mg/kg.Islets were isolated using digestion by ductal injection of collagenase,and purified by centrifugation of Ficoll density gradient.The cells with high purity,which had DC characteristic of morphology and function,could be generated by cytokines-induced.The DC were transfected with the recombinant adenovirus Adv-CTLA4Ig and transplanted into diabetic recipients' renal subcapsule with pancreatic islets at the same time.The expression of CTLA4Ig gene in experimental and control groups was detected by reverse transcriptase polymerase chain reaction and Dot-ELISA and immunohistochemistry methods.The level of blood glucose was determined before and after transplantation and the survival time of recipients was recorded.Results The expression of CTLA4Ig gene was detected in experimental group,but not detectable in control group.The mean normglymia time in the experimental group was (17.3± 2.4) days,significantly longer than in the control group (10.1± 1.5) days and vacant control group (8.3± 1.2) days.The survival time in experimental group was (34.5± 3.4) days,significantly longer than in the control group (14.7± 2.3) days and vacant control group (11.2± 1.4) days.Conclusion DC expressing CTLA4Ig gene could induce immunologic tolerance in pancreatic islet transplantation,and prolong significantly survival time of pancreatic islet allografts.

关 键 词：细胞毒性T淋巴细胞相关抗原4 免疫球蛋白基因 同供体 移植物存活 树突状细胞 CTLA4IG基因 转染 逆转录一聚合酶链反应 密度梯度离心法 DC细胞 免疫组织化学 移植免疫耐受 对照组 糖尿病模型 腹腔内注射 重组腺病毒 ELISA 实验组 胰岛移植物 SD大鼠 链尿菌素 胶 

分 类 号：R657.5[医药卫生—外科学]