利用SUMO表达系统高效可溶表达人白细胞介素15及其活性研究  

Use of the SUMO expression system to efficiently express the soluble human interleukin-15 and the activity study

作　　者：白福良[1] 田辉[1] 牛泽杉[1] 于引航[1] 王文飞[1] 周兵[1] 李德山[1] 

机构地区：[1]东北农业大学生命科学学院生物制药教研室,哈尔滨150030

出　　处：《中国免疫学杂志》2013年第7期689-693,共5页Chinese Journal of Immunology

摘　　要：目的:探讨重组人白细胞介素15(rhIL-15)的表达条件、纯化工艺并进行生物活性鉴定。方法:采用RT-PCR方法从正常成人外周血淋巴细胞中扩增出人白细胞介素15(hIL-15)cDNA,克隆到pSUMO Vector中,构建rpSUMOhIL-15重组质粒。将转化重组质粒的工程菌诱导表达,超声破碎后取菌液上清,经过亲和层析、酶切、亲和层析得到成熟蛋白。高效液相色谱进行纯度分析;利用rhIL-15促进T细胞增殖、rhIL-15诱生的NK细胞杀伤肿瘤细胞的能力鉴定其生物活性;荷瘤鼠实验研究rhIL-15抗肿瘤作用。结果:经纯化后得到rhIL-15,其分子量大小为14.4 kD,蛋白纯度在95%以上,具有显著刺激T细胞增殖和NK细胞杀伤肿瘤细胞效果。动物实验结果显示其具有明显的抗肿瘤作用。结论:经过纯化获得高纯度和高生物学活性的rhIL-15,且具有明显增强NK细胞杀伤、刺激T细胞增殖和抗肿瘤的活性,为细胞因子联合其他方法治疗肿瘤等疾病提供了实验基础。Objective：To investigate the expression,purification and bioactivity detection of the recombinant human IL-15 protein（rhIL-15）.Methods： The entire human Interleukin-15（hIL-15） coding region was amplified from human peripheral blood lymphocytes by RT-PCR and subcloned into the pSUMO vector to form rpSUMOhIL-15.The rhIL-15 was purified from rhIL-15-producing bacteria followed by affinity chromatography,protease enzyme digestion,and affinity chromatography.The purified rhIL-15 was analyzed by HPLC（High Performance Liquid Chromatography） technology.The bioactivities of the purified rhIL-15 were detected by T cell proliferation and NK cytotoxicity tests.The anti-tumor effect of rhIL-15 was studied in tumor-bearing mice.Results： The purified rhIL-15 with a purity of 95% was obtained and molecular weight was 14.4 kD evaluated by SDS-PAGE.The purified rhIL-15 significantly enhanced NK cytotoxicity and stimulated proliferation of T cells.Animal experiments showed that rhIL-15 significantly inhibited tumor growth.Conclusion： The bioactive rhIL-15 has been obtained.The rhIL-15 protein enhances killing activities of NK cells for HepG2 and B16-F10 cells,stimulates the proliferation of human T cells and inhibits tumor growth.This study provides experimental evidence for treatment of cancers or other diseases with rhIL-15.

关 键 词：rpSUMOhIL-15 融合蛋白 T细胞增殖 NK细胞杀伤活性 抑制肿瘤作用 

分 类 号：R915[医药卫生—微生物与生化药学]