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重组人ICOS蛋白在大肠杆菌中的表达及其对B细胞活性的作用(英文)
Expression of Recombinant Human ICOS and in Vitro Characterization of Its Bioactivity on B Lymphocytes
【摘要】 ICOS (induciblecostimulator)是一种表达在活化T细胞上的新型共刺激分子 ,它的配体GL5 0组成性表达在B细胞及巨噬细胞上。ICOS可以促使Th2细胞的分化 ,能促进IL 4、IL 10的分泌。从扁桃体中克隆到ICOS基因 ,构建了PET ICOS重组载体 ,并在大肠杆菌中得到高效表达 ,经SDS电泳及Western印迹法鉴定 ,所获得的蛋白质为特异性目的蛋白质。继而利用ICOS联合IL 10以及PWM驱动的B细胞体外培养系统 ,分析了ICOS在B细胞生长及抗体产生中的作用。结果表明 ,ICOS在PWM体外培养体系中 ,能有效地促进B细胞生长以及协同IL 10增加IgG的分泌。
【Abstract】 Inducible costimulator (ICOS) is a novel costimulatory molecule expressed in activated T cell and has critical regulation effect on special immune response. In this study, the cDNA encoding human ICOS was cloned from activated tonsil cells via RT PCR, and was expressed in E. coli on pET28 expression vector. The recombinant ICOS protein expressed from E. coli showed a molecular weight of 14 kD on SDS polyacrylamide gel electrophoresis and was further confirmed by Western blot. In presence of IL 10, the purified rhICOS significantly increased in vitro B cell growth stimulated by pokeweed mitogen (PWM), and enhanced the secretion of IgG from B cells.
【Key words】 costimulatory molecules; ICOS protein; E. coli; antibody;
- 【文献出处】 生物化学与生物物理学报 ,Acta Biochimica Et Biophysica Sinica , 编辑部邮箱 ,2003年07期
- 【分类号】Q78
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