鳗弧菌L-18株的限铁条件和铁调节外膜蛋白的免疫效果研究
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科技部科研院所社会公益研究专项


Iron-restricted condition and the immune efficacy of iron-regulated outer membrane proteins of Vibrio anguillarum L-18
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    摘要:

    利用Western-blot技术,对鳗弧菌L-18株的OMPs和IROMPs的免疫反应性进行了分析,并通过免疫牙鲆比较二者免疫保护力的差异,以期从IROMPs方向探索开发鳗弧菌新型鱼用疫苗,并为研究其免疫原理提供重要的科学依据。结果表明,培养基中加入不同浓度的联铁剂,鳗弧菌L-18株的生长和铁载体的表达出现规律性变化,其中加入100~130μmol·L-1 2,2’-联吡啶为最适合限铁条件,此时铁载体的表达量最高且生长受抑制程度较小。在此条件下,菌株表达出74.3ku和77.4ku的IROMPs,其中77.4ku蛋白具有免疫反应性,为重要抗原。用全菌灭活疫苗、OMPs、IROMPs分别腹腔注射免疫牙鲆7周后攻毒, IROMPs疫苗的相对免疫保护力达到75.9%,远高于OMPs疫苗的51.8%。相对于OMPs,IROMPs的抗体效价显著提高,接近全菌疫苗组的水平,而血清杀菌活力没有显著变化。 关键词:鳗弧菌;铁调节外膜蛋白;牙鲆

    Abstract:

    Vibrio anguillarum is a familiar pathogen of marine fishes, which express a kind of specific iron-uptake system when infection. The iron-regulated outer membrane proteins (IROMPs) of Vibrio anguillarum have been learned for its expression and composition, but not for the immune efficacy on fish yet. The immunoreactivity of outer membrane proteins (OMPs) and IROMPs of Vibrio anguillarum L-18 were analyzed with Western-blot in this study, and the difference of immune efficacy of OMPs and IROMPs was determined through immunizing flouder Paralichthys olivaceus. We found that the growth and the output of siderophore of L-18 were associated with the concentration of 2, 2’-dipyridyl (a kind of iron chelator) in media TSB. 100~130μmol·L-1 2, 2’-dipyridyl in TSB was affirmed to be the most proper iron-restricted condition, in which the growth of L-18 hadn’t been restrained strongly and the output of siderophore had reached to the tiptop. Under this iron-restricted condition, L-18 expressed 74.3ku and 77.4ku IROMPs. The 77.4ku protein, which was considered to be a important antigen, showed immunoreactivity clearly in Western-blot analyse. After immunization through intraperitoneal injection for 7 weeks , the cumulative mortality of P. olivaceus showed different values among formalin-killed whole-cell vaccine group, OMPs group and IROMPs group. The relative percent survival (RPS) of IROMPs vaccine reached to 75.9%, higher than 51.8% of OMPs vaccine significantly. The antibody titre of IROMPs group advanced significantly compared with OMPs group, nearly the same as the whole-cell group, but the survival index (SI) of it in serum had no obvious change. Key words: Vibrio anguillarum ; IROMPs ; Paralichthys olivaceus

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高磊.鳗弧菌L-18株的限铁条件和铁调节外膜蛋白的免疫效果研究[J].水产学报,2007,31(4):472~476

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  • 收稿日期:2007-07-16
  • 最后修改日期:2007-07-16
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  • 在线发布日期: 2007-07-17
  • 出版日期: 2007-07-17
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