Expression of recombinant human ICOS and in vitro characterization of its bioactivity on B lymphocytes.

Zhong-Bin Deng, Chang-Ming Lu, Wei-Da Huang, Li-Qin Shen, Wei Zhu, Hong-Bing Ma, Pan-Sheng Fan, Xue-Guang Zhang
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引用次数: 0

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.

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重组人ICOS的表达及其对B淋巴细胞生物活性的体外表征。
诱导共刺激因子(Inducible costimulator, ICOS)是一种表达于活化T细胞中的新型共刺激分子,对特定的免疫应答具有重要的调节作用。本研究通过RT-PCR从活化的扁桃体细胞中克隆出编码人ICOS的cDNA,并在大肠杆菌pET28表达载体上进行表达。从大肠杆菌中表达的重组ICOS蛋白经sds -聚丙烯酰胺凝胶电泳证实分子量为14 kD,并经Western blot进一步证实。在IL-10的作用下,纯化后的rhICOS可显著促进美洲商陆丝裂原(PWM)诱导的体外B细胞生长,增强B细胞IgG的分泌。
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