五指山小型猪单个核细胞表面α Gal抗原表达差异的分析  

Variations Analysis of α1,3 Gal Level in Wuzhishan Miniature Porcine

作　　者：陈苹[1] 刘羿男[1] 王卫平[1] 周春燕[1] 马康涛[1] 

机构地区：[1]北京大学基础医学院生物化学与分子生物学系,北京100191

出　　处：《中国生物化学与分子生物学报》2010年第3期254-259,共6页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家高技术研究发展计划(No.863计划;2003AA205090);北京大学985项目资助~~

摘　　要：利用猪的器官为挽救脏器终末衰竭病人而进行的异种移植是有效救治病人的途径之一.由于猪细胞表面存在的α1,3半乳糖会引起超急性免疫排斥反应,导致移植器官最终被移植受体排斥.除人类和旧大陆猴外,所有哺乳动物细胞表面都有αGal半乳糖表达,但个体间表达量存在显著差异.为研究近交系五指山小型猪细胞表面α1,3半乳糖表达的个体差异,本实验利用FITC-isolectin进行荧光标记,通过流式细胞术以及激光共聚焦分析,对14例5月龄猪样本外周血单个核细胞表面的α1,3半乳糖水平进行检测.利用猪肾上皮PK15细胞确定FITC-isolectin标记的最适浓度为25ng/μl(细胞标记效率大于85%).结果显示,14头猪外周血单个核细胞表面α1,3半乳糖的表达差异范围在1.25～2.09倍之间,远低于普通非近交系猪的个体差异.本研究为利用近交系五指山小型猪作为异种器官移植的研究材料提供了基础数据.The expression of α 1,3 Gal in inbred strain Wuzhishan miniature porcine was determined by flow cytometry and confocal analysis.The porcine peripheral blood mononuclear cells（PBMCs） were isolated from heparinised whole blood of 5-month old porcine by density gradient centrifugation and then incubated with FITC-isolectin.The specificity of FITC-isolectin binding to α 1,3 Gal was tested in porcine kidney epithelial PK15 and human HEK293 cells.No detectable fluorescence was observed in HEK293 cells with 100 ng/μl FITC-isolectin treatment,while the fluorescence was detected in PK15 cells with only 1.560 ng/μl FITC-isolectin treatment.The optimal concentration of FITC-isolectin was determined as 25 ng/μl（labeling rate85%） for PK15 cells.The samples were analyzed on a Calibur FACScan and BX51 Laser scanning confocal microscope.The α 1,3 Gal levels were found to display a 1.25～2.09 folds variation in the samples examined,indicating the variation of α 1,3 Gal expression between individual porcine appeared to be limited.These findings may further validate the inbred Wuzhishan miniature porcines as the candidate organ donors for xenotransplantation.

关 键 词：异种抗原 α1 3半乳糖 五指山小型猪 表达差异 

分 类 号：Q533[生物学—生物化学]