人重组趋化素样因子1在果蝇S2细胞中的表达和功能研究  

Expression and functional study of human recombinant chemokine-like factor 1 in Drosophila S2 ceils

作　　者：张颖妹[1] 李婷[1] 娄雅欣[1] 韩文玲[1] 马大龙[1] 

机构地区：[1]北京大学人类疾病基因研究中心,北京大学医学部免疫学系,100083

出　　处：《中华微生物学和免疫学杂志》2008年第6期481-485,共5页Chinese Journal of Microbiology and Immunology

基　　金：高等学校全国优秀博士论文作者专项资金（FANEDD,200257）;国家自然科学基金（30671906）

摘　　要：目的在果蝇S2细胞中表达人趋化素样因子1（chemokine-like factor1，CKLF1），并对其分泌形式进行功能研究。方法构建pMT／V5-His—CKLF1表达质粒，转染S2细胞，筛选并鉴定阳性细胞克隆，用兔抗人CKLF1多肽抗体对其培养上清进行Western blot检测，并分析其趋化活性和促C2C12细胞增殖活性。结果RT—PCR证明CKLF1在S2细胞中高效转录；通过Western blot在细胞培养上清中可检测到表达的重组CKLF1蛋白；其对人外周血中性粒细胞和淋巴细胞有较弱的趋化活性，对C2C12细胞具有增殖促进作用。结论在果蝇S2细胞中成功表达了人重组CKLF1，并证实其存在分泌形式且具有趋化活性和促C2C12细胞增殖活性。Objective To express human chemokine-like factor 1 （ CKLF1 ） in Drosophila S2 cells and study its function. Methods The pMT/VS-His-CKLF1 expression plasmid was constructed and transfected into Drosophila S2 cells. The positive clones were selected through PCR and RT-PCR. The culture medium was analyzed by Western blot with anti-CKLF1 polyclonal antibody. Chemotaxis and MTF assays on human peripheral blood and C2C12 cells, respectively, were then carried out with the medium. Results CKLF1 was transcribed efficiently in S2 cells. The expressed CKLF1 protein could be detected in the culture supernatant by Western blot, which showed weak chemotactic activity on both human peripheral blood neutrophils and lymphocytes as well as enhancing effect on the proliferation of C2C12 cells. Conclusion CKLF1 was expressed successfully in Drosophila S2 cells and secreted into the culture medium. The recombinant CKLF1 expressed in Drosophila cells can chemoattract leucocytes and promote the proliferation of C2C12 cells.

关 键 词：趋化素样因子1 趋化因子 C2C12细胞 

分 类 号：R392[医药卫生—免疫学]