禽多杀性巴氏杆菌C48-3株外膜蛋白H的致病作用

doi:10.13523/j.cb.20140605

中国生物工程杂志

2014, Vol. 34

Issue (06): 31-39 DOI: 10.13523/j.cb.20140605

研究报告

禽多杀性巴氏杆菌C48-3株外膜蛋白H的致病作用

韦东¹, 吾鲁木汗·那孜尔别克¹, 段世雄¹, 严芳², 恩特马克·布拉提白¹

1. 吉首大学生物资源与环境科学学院吉首 416000;
2. 吉首市畜牧水产局动物疫病预防控制中心吉首 416000

Role of Outer Membrane Protein H in the Pathogenesis of Avian Pasteurella multicida Strain C48-3

WEI Dong¹, WULUMUHAN Nazierbieke¹, DUAN Shi-xiong¹, YAN Fang², ENTOMACK Borrathybay¹

1. College of Biology and Environmental Sciences, Jishou University, Jishou 416000, China;
2. Animal Disease Prevention and Control Center, Bureau of Animal Husbandry and Fishery Industries of Jishou City, Jishou 416000, China

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摘要：

目的：探讨禽多杀性巴氏杆菌外膜蛋白H（OmpH）的致病作用。方法：用大肠杆菌表达系统表达强毒株C48-3的重组蛋白OmpH，亲和层析法纯化N端带有6个组氨酸标签的重组OmpH，通过皮下注射兔制备抗OmpH抗血清，用生物学功能实验比较野生株C48-3、突变株ΔompH和互补株C48-3C的粘附能力、血清抵抗性和抗吞噬作用。结果：与野生株和互补株相比，突变株对CEF细胞的粘附能力显著降低，而抗OmpH抗体显著抑制野生株和互补株对CEF细胞的粘附，但该抗血清不影响突变株的粘附能力。野生株和互补株在鸡血清中的存活率显著高于突变株，但灭活鸡血清处理不影响它们的存活率。小鼠腹腔巨噬细胞对突变株的吞噬能力显著高于野生株和互补株，而抗OmpH抗体增强巨噬细胞对野生株和互补株的吞噬能力，但该抗体不影响巨噬细胞对突变株的吞噬能力。结论：OmpH是禽巴氏杆菌的致病因子，它在该菌对宿主的感染与致病过程中发挥重要的作用。

关键词： 禽多杀性巴氏杆菌; OmpH; 粘附; 血清抵抗性; 抗吞噬作用

Abstract:

The previous study has indicated that the targeted inactivation of ompH gene decreased the lethality of serotype A Pasteurella multocida C48-3 in mice after intraperitoneal injection. The ompH gene was cloned from P. multocida C48-3 and expressed in Escherichia coli M15, and immune serum was generated by immunizing rabbit with the purified recombinant protein OmpH. Differences of adhesion ability, serum resistance and antiphagocytic activity between the wild-type strain C48-3, mutant strain ΔompH and its complemented strain C48-3C were compared by biological experiments. The adhesion of wild-type strain and complemented strain to CEF cells was inhibited significantly by treatment with immune sera as compared to the treatment with normal serum. The mutant strain was sensitive to the bactericidal action of chicken serum, whereas the wild-type strain and complemented strain were both resistant. The mutant strain was readily taken up by mouse peritoneal macrophages in the normal rabbit serum, whereas the wild-type strain and complemented strain were both resistant. In addition, the rabbit antiserum against OmpH promoted the phagocytosis of C48-3 and C48-3C by mouse peritoneal macrophages, but mutant strain was not affected. These results demonstrate that the OmpH is a major virulence factor of P. multocida.

Key words: Avian Pasteurella multocida OmpH Adhesion serum resistance Antiphagocytosis

收稿日期: 2014-03-31 出版日期: 2014-06-25

ZTFLH:

Q819

基金资助:

国家自然科学基金资助项目（30972206，31360613）

通讯作者: 恩特马克·布拉提白 E-mail: etmkb@jsu.edu.cn

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引用本文:

韦东, 吾鲁木汗·那孜尔别克, 段世雄, 严芳, 恩特马克·布拉提白. 禽多杀性巴氏杆菌C48-3株外膜蛋白H的致病作用[J]. 中国生物工程杂志, 2014, 34(06): 31-39.

WEI Dong, WULUMUHAN Nazierbieke, DUAN Shi-xiong, YAN Fang, ENTOMACK Borrathybay. Role of Outer Membrane Protein H in the Pathogenesis of Avian Pasteurella multicida Strain C48-3. China Biotechnology, 2014, 34(06): 31-39.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20140605 或 https://manu60.magtech.com.cn/biotech/CN/Y2014/V34/I06/31

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