表达不同致病型S基因重组鸡传染性支气管炎病毒的细胞嗜性研究

发布时间：2018-04-08 11:05

本文选题：传染性支气管炎病毒(IBV)　切入点：刺突蛋白(S)　出处：《病毒学报》2017年05期

【摘要】：鸡传染性支气管炎病毒(IBV)在体外进行细胞培养比较困难,并且不同来源毒株在易感性上也存在差异。为揭示IBV适应CK细胞培养的分子基础,优化其细胞培养技术,本研究首先对H120疫苗株和IBYZ分离株的分子克隆株rH120和rIBYZ,在鸡胚、鸡气管环以及原代鸡肾细胞(CK)上培养时组织细胞的嗜性差异进行了比较,结果发现rIBYZ与rH120在CK上的亲嗜性存在显著差异。在此基础之上,课题组借助前期建立的IBV反向遗传操作系统,将H120疫苗株和IBYZ株的S基因进行交叉替换,成功拯救获得2株重组病毒rH120-S/YZ和rIBYZ-S/H120。通过比较不同毒株感染CK细胞后的病变、特异性免疫荧光的强度以及病毒的增殖曲线,表明刺突蛋白(S)在IBV适应CK细胞过程中发挥决定性作用。本文揭示了S蛋白对细胞嗜性的影响,为进一步探索IBV适应细胞的分子机制奠定了基础。
[Abstract]:Chicken infectious bronchitis virus (IBV) was difficult to culture in vitro, and the susceptibility of different strains was different.In order to reveal the molecular basis of IBV adapting to CK cell culture and to optimize its cell culture technique, the molecular clones rH120 and rIBYZ of H120 vaccine and IBYZ isolate were first studied in chicken embryo.The difference of tissue cell tropism between rIBYZ and rH120 on CK was found to be significantly different from that of rH120 in the culture of chicken trachea ring and primary chicken kidney cell (CK). The results showed that there was a significant difference in the cytotropism between rIBYZ and rH120 on CK.On the basis of the above, the S gene of H120 vaccine strain and IBYZ strain were cross-replaced by the IBV reverse genetic operating system, and two recombinant viruses, rH120-S/YZ and rIBYZ-Sr H120, were successfully saved.By comparing the pathological changes, the intensity of specific immunofluorescence and the curve of virus proliferation of CK cells infected with different virulent strains, it is concluded that Sus) plays a decisive role in the adaptation of IBV to CK cells.In this paper, the effect of S protein on cell eosinophilia was revealed, which laid a foundation for further exploring the molecular mechanism of IBV adaptation to cells.
【作者单位】：中国农业科学院家禽研究所;
【基金】：国家自然科学基金青年基金(项目号:31602091),题目:S蛋白介导的鸡传染性支气管炎病毒适应CK细胞的分子基础研究;国家自然科学基金面上项目(项目号:31572524),题目:传染性支气管炎病毒胚化致弱株毒力减弱的分子机制研究江苏省科技支撑计划(项目号:BE2012369),题目:鸡传染性支气管炎基因重组减毒活疫苗的研制扬州市自然科学基金(项目号:YZ2014029),题目:嵌合流行株S基因重组鸡传染性支气管炎病毒疫苗候选株的构建及其免疫原性研究~~
【分类号】：S852.65

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