Galeetin-9（Tim-3L）显著减弱小鼠移植排斥  

Galectin-9（Tim-3L） significantly attenuates aUogeneic immune response in mice

作　　者：何文涛[1] 袁劲[1] 方泽民[1] 汪峰[1] 徐逸[1] 周鸿敏[1] 高英[1] 张维娜[1] 王璐[1] 陈忠华 

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,教育部/卫生部重点实验室,武汉430030

出　　处：《中华微生物学和免疫学杂志》2009年第1期5-10,共6页Chinese Journal of Microbiology and Immunology

基　　金：基金项目：国家自然科学基金资助项目（30671989）;国家重点基础研究发展规划（973计划）（2009CB522407）

摘　　要：目的探究Galectin-9（即Tcell immunoglobulin domain and mucin domain protein-3 lig-and，Tim-3L）的真核定位与功能，及其对同种异基因排斥反应的影响。方法PCR扩增Galectin-9片段，插入到pEGFP—N1质粒，转染中国仓鼠卵巢癌细胞系（CHO）行G418筛选，分选增强型绿色荧光蛋白（EGFP）阳性细胞。免疫组化检测Galectin-9的表达，Alamar Blue法检测稳定转染Galectin-9的CHO细胞及其新鲜培养上清对脾细胞增殖的影响，ELISA检测IL-2水平。给予BALB／c→C57BL／6心脏移植受鼠Galectin-9蛋白100μg×7d。结果Galectin-9表达于细胞浆，稳定转染Galectin-9-EGFP的CHO细胞上清抑制淋巴细胞增殖和IL-2的产生。上述效应可被Tim-3-Fc融合蛋白逆转。Galectin-9干预的心脏移植物中位生存时间明显延长[（22．7±1．2）d，（7．2±0．4）d）]。结论真核细胞中Galectin．9可通过分泌形式负调节同种异基因免疫；Galectin-9有望成为新型的抗排斥药物。Objective To explore the subcellular localization of Galectin-9 and its effect on allogeneic immune response. Methods The plasmid pEGFP-N1 was inserted with Galectin-9 fragment which was amplified from pBKCMV-Galectin-9 by PCR. The recombinant plasmid was then transfected into CHO cells using JetPEl in vitro. The cells were cultured in G418 selecting medium to obtain the stably-transfected cells. The transcription and expression of Galectin-9 gene were verified by immunohistochemical staining and RT-PCR. The solid-phase transgenic CHO cells or freshly-cultured supernatant was added into the mixed lymphocyte response system to detect the inhibitory effect of Galectin-9. Galectin-9 protein was administered intraperitoneally for 7 d consecutively. Results The expression of Galectin-9 was localized in the cytosol of CHO. The allogeneic mix lymphocyte proliferation was dose-dependently inhibited by the freshly-cultured su- pernatant from stably-transfected CHO cells. Furthermore, the supernatant from stably-transfected CHO cells dose-dependently inhibited the level of IL-2. The inhibitory effect could be reversed by Tim-3-Fc blocking. Administration of Galectin-9 significantly prolonged the survival of allogeneic cardiac transplants [ （22.7 ± 1.2） d vs （7.2 ± 0.4） d） ]. Conclusion Galectin-9 may be secreted in physical situation to exert its im- munomodulatory function on allogeneic immune response. Furthermore, Galectin-9 may be a novel therapeutic drug in transplant medicine.

关 键 词：TIM-3 GALECTIN-9 免疫抑制 小鼠 

分 类 号：R392.4[医药卫生—免疫学] R392.11[医药卫生—基础医学]