人4-1BBL胞外区基因的克隆与表达研究  

Cloning and Expression of the Extracellular Domain of 4-1BBL

作　　者：姜文国[1] 熊冬生[1] 邵晓枫[1] 王金宏[1] 许元富[1] 刘芳[1] 郭红星[1] 朱祯平[1] 杨纯正[1] 

机构地区：[1]中国医学科学院中国协和医科大学血液学研究所实验血液学国家重点实验室,天津300020

出　　处：《生物工程学报》2005年第5期703-707,共5页Chinese Journal of Biotechnology

基　　金：国家高技术研究发展计划项目(863)资助(No.2003AA215080);天津市重大科技攻关经费资助(No.003119511)。~~

摘　　要：用RTPCR方法从人单核THP1细胞系克隆人41BBL胞外区基因,将其重组到pAYZ表达载体中,构建成人41BBL胞外区基因表达载体。将该载体转化大肠杆菌16C9,获得稳定表达,表达产物主要以可溶性状态存在;SDSPAGE和Westernblot分析显示,其分子量约为22kD,与预期结果一致。这是首次在大肠杆菌中获得41BBL胞外区可溶性表达。生物学活性检测显示41BBL对于维持T淋巴细胞系因子释放非常有益,同时PI单染表明它能抑制Jurkat细胞的凋亡。这将在抗肿瘤免疫治疗中具有潜在应用前景。RT-PCR was used to clone DNA fragment of the extracellular domain of 4-1BBL from human THP-1 cells （human monocyte）, and the expression vector pAYZ4-1BBL was constructed by cloning the extracellular domain of 4-1BBL into the expression vector pAYZ. The extracellular domain of 4-1BBL was expressed in E. coli 16C9 and purified by affinity chromatography, SDS-PAGE and Western blot analysis showed that the relativae molecular weight of soluble 4-1BBL is 22kD which was consistent with the theoretically predicted value. So far as we know, it is the first time that the soluble expression of 4-1BBL in E. coli.was achieved 4-1BBL induced a significant release of IL-2 in stimulated Jurkat cells after 48h incubation, especially in the presence of tumor cell. At the same time the apoptosis level of Jurkat cell reduce more than 50%. In conclusion,4-1BBL may be useful in cancer immunotherapy.

关 键 词：4-1BBL 协同刺激分子 原核表达 

分 类 号：R392.11[医药卫生—免疫学]