人髓系细胞触发受体-1模拟多肽的筛选和鉴定  

Screening and identification of human triggering receptor expressed on myeloid cells-1 mimetic peptides

作　　者：杨其霖[1] 黄红川[1] 张彦峰[1] 何为群[1] 刘晓青[1] 黎毅敏[1] 

机构地区：[1]广州医学院第一附属医院呼吸疾病研究所ICU,广州市510120

出　　处：《中华老年多器官疾病杂志》2008年第6期506-510,共5页Chinese Journal of Multiple Organ Diseases in the Elderly

基　　金：广东省自然科学基金资助课题(资助号:06022087)

摘　　要：目的寻找能特异性与人髓系细胞触发受体-1(TREM-1)蛋白结合并抑制其传导通路的多肽。方法克隆、表达和纯化TREM-1功能区蛋白,并以之为诱饵蛋白,筛选随机噬菌体展示肽库。经过4轮生物淘洗,通过ELISA方法和单核细胞ELISA方法分析噬菌体克隆与TREM-1蛋白的亲和力。化学合成模拟多肽,检测其对盲肠结扎穿刺(CLP)小鼠的治疗效果。结果成功找到5种噬菌体克隆,ELISA法和细胞ELISA均阳性。应用化学方法合成的模拟多肽HYGMTHPNTMSH能降低CLP小鼠的死亡率。结论模拟多肽HYGMTHPNTMSH对脓毒症小鼠有保护作用。Objective To find mimetic peptides which bind specifically with triggering receptor expressed on myeloid cells-1 （TREM-1） and inhibit the TREM-1 pathway. Methods The functioning region of TREM-1 protein was cloned, expressed and purified. With the recombinant protein of TREM-1 as the target molecule, the mimetic peptides were screened from a random phage display peptide library. Four rounds of biopanning were carried out, and enzyme linked immunosorbent assay （ELISA） and monocyte cell ELISA were used to evaluate the binding between the phage clone and TREM-h Then, chemical synthesis of the mimetic peptide was carried out, and its therapeutic effect on cecal ligation and puncture （CLP） in mice was investigated. Results Five phage clones from the fourth round of biopanning were selected at random, all of them were positive in sandwich ELISA and monocyte cell ELISA. Administration of the chemically synthesized mimetic peptide HYGMTHPNTMSH reduced the mortality of CLP mice. Conclusion Mimetic peptide HYGMTHPNTMSH can protect the mice wtih sepsis from death.

关 键 词：髓系细胞触发受体-1 模拟多肽 筛选随机噬菌体展示肽库 

分 类 号：R446.113[医药卫生—诊断学] R392.9[医药卫生—临床医学]