| 重组IL-18BP拮抗重组鸡IL-18刺激外周血单核细胞产生IFN-γ的作用 |
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| 引用本文： | 刘文强,刘桂芹,秦绪岭,王桂英.重组IL-18BP拮抗重组鸡IL-18刺激外周血单核细胞产生IFN-γ的作用[J].微生物学报,2010,50(4):506-511. |
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| 作者姓名： | 刘文强 刘桂芹 秦绪岭 王桂英 |
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| 作者单位： | 聊城大学农学院,聊城大学畜禽疫病防制技术研究所,聊城,252059 |
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| 基金项目： | 基金项目：国家自然科学基金（30800130）；山东省中青年科学家奖励基金（2006BS02012）；聊城大学博士基金（31805） |
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| 摘 要： | 【目的】白细胞介素-18通过激活Th1细胞和NK细胞产生IFN-γ而发挥关键的免疫调节作用。人和小鼠分泌的白细胞介素-18结合蛋白(IL-18BP)可以拮抗其活性。推测在鸡痘病毒基因组中也含有IL-18BP基因的同源物,对其表达的蛋白质进行了活性鉴定,为拮抗IL-18主导的疾病提供理论依据。【方法】根据鸡痘疫苗病毒的基因组序列设计特异性引物,使用PCR方法从中分离cIL-18BP基因,将该基因克隆到酵母表达载体pPICZαA中,甲醇诱导后在酵母GS115中进行表达。对表达的重组蛋白进行了活性鉴定。【结果】从鸡痘病毒中克隆到cIL-18BP基因,SDS-PAGE鉴定了该基因在酵母系统中的高效表达。ELISA检测表明纯化后的cIL-18BP与重组鸡(c)IL-18发生特异性结合;通过测定IFN-γ的浓度,表明该蛋白具有拮抗IL-18刺激外周血单核细胞(PBMCs)和MSB1细胞分泌IFN-γ的活性。【结论】实验表明,cIL-18BP通过抑制cIL-18刺激相关免疫细胞分泌IFN-γ而发挥对IL-18的拮抗作用,敲除该基因可能有助于研制更安全和高效的鸡痘疫苗。
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| 关 键 词： | 关键词：鸡痘病毒 IL-18结合蛋白 活性 |
| 收稿时间： | 2009/8/16 0:00:00 |
| 修稿时间： | 2010/1/20 0:00:00 |
Recombinant cIL-18-binding protein as an antagonist to cIL-18 enhanced PBMCs secreting IFN-γ |
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| Wenqiang Liu,Guiqin Liu,Xuling Qin and Guiying Wang.Recombinant cIL-18-binding protein as an antagonist to cIL-18 enhanced PBMCs secreting IFN-γ[J].Acta Microbiologica Sinica,2010,50(4):506-511. |
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| Authors: | Wenqiang Liu Guiqin Liu Xuling Qin and Guiying Wang |
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| Institution: | Agricultural College of Liaocheng University, Prevention and Control Technology Institute of Animal Disease in Liaocheng University, Liaocheng 252059, China;Agricultural College of Liaocheng University, Prevention and Control Technology Institute of Animal Disease in Liaocheng University, Liaocheng 252059, China;Agricultural College of Liaocheng University, Prevention and Control Technology Institute of Animal Disease in Liaocheng University, Liaocheng 252059, China;Agricultural College of Liaocheng University, Prevention and Control Technology Institute of Animal Disease in Liaocheng University, Liaocheng 252059, China |
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| Abstract: | Objective]Interleukin-18(IL-18) is a proinflammatory cytokines that plays a key role of immune regulation through activating the Th 1 cell and NK cell secreting interferon-γ(IFN-γ) .IL-18-binding proteins(IL-18BP) secreted in human and mouse can antagonise the activity of IL-18.The homologue gene of IL-18BP from fowlpox virus genome was predicted and identified of its expressed protein, to use as an antagonist to IL-18 guiding disease.Methods]The cIL-18BP gene was isolated from the genome of fowlpox vaccine virus by PCR through a pair of special primers and cloned into vectors pPICZαA, then expressed in yeast GS115.The biologic activity of recombinant proteins binding with cIL-18 was measured, inhibiting the activity of cIL-18 enhancing relative cells secreting IFN-γ was confirmed through ELISA.Results]The cIL-18BP gene was cloned from fowlpox virus and acquired high performance expression in yeast systems induced with methanol identified by SDS-PAGE.Recombinant cIL-18BP purified can bind specifically with recombinant cIL-18 determined by ELISA test.cIL-18BP can antagonise the activity of cIL-18 with determining the IFN-γ concentration secreting in PBMCs and MSB1 cells stimulated by cIL-18 respectively.Conclusion]The experiment indicates that recombinant cIL-18BP can inhibit the activity of cIL-18 stimulating related immune cells secreting IFN-γ.Deletion of the cIL-18BP from virus may improve the safety and immunogenicity of fowlpox virus vaccine. |
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| Keywords: | Keywords: fowlpox virus IL-18BP activity |
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