Center for Proteomics, State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China.
Fish Shellfish Immunol. 2010 Nov;29(5):810-6. doi: 10.1016/j.fsi.2010.07.027. Epub 2010 Jul 24.
Vibrio alginolyticus is the etiological agent that causes great losses in aquacultures and clinical emanating cases in humans. Identification of highly efficient vaccine candidates to control V. alginolyticus infection has been highly concerned since vaccines offer a powerful approach to provide efficient protection from bacterial infections. In the present study, we firstly investigated the altered outer membrane proteins (OM proteins) of V. alginolyticus in response to NaCl concentrations and iron limitation using Western blotting, and then identified the protective activity of these altered OM proteins by bacterial challenge post immunization. Ten OM proteins were differentially expressed in response to the osmolarity changing or/and iron limitation, in which VA2212, OmpV, VPA1186, OmpU, VPA1644, VA1061, VA1631 and VPA0860 were markedly altered in response to osmolarity, and VPA1186, OmpU, OmpV, VA0449, VPA0860, VPA1435 and VA1631 were determined to be iron-limited responsive proteins. Out of the ten OM proteins, VA1061, OmpU, VPA1435 and VPA0860 could be effective vaccine candidates against infection by V. alginolyticus in vivo. Further results indicated that VA1061 and VPA0860 were dominant antigens and could stimulate hosts to produce stronger antibody response than other two in live or inactivated whole-cell vaccines. These results not only expand knowledge on osmolarity-, iron-responsive proteins, but also provide a valuable strategy for identify protective proteins suitable for use in vaccine development.
溶藻弧菌是水产养殖和人类临床感染的重要病原体。由于疫苗提供了一种有效的细菌感染保护方法,因此,鉴定高效的疫苗候选物来控制溶藻弧菌感染受到了高度关注。在本研究中,我们首先使用 Western blot 技术研究了溶藻弧菌在外膜蛋白(OM 蛋白)在盐度和铁限制下的变化,然后通过免疫后细菌攻毒来鉴定这些改变的 OM 蛋白的保护活性。有 10 种 OM 蛋白对渗透压变化和/或铁限制有差异表达,其中 VA2212、OmpV、VPA1186、OmpU、VPA1644、VA1061、VA1631 和 VPA0860 对渗透压的变化有明显改变,而 VPA1186、OmpU、OmpV、VA0449、VPA0860、VPA1435 和 VA1631 是铁限制响应蛋白。在这 10 种 OM 蛋白中,VA1061、OmpU、VPA1435 和 VPA0860 可以作为有效的疫苗候选物,预防溶藻弧菌感染。进一步的结果表明,VA1061 和 VPA0860 是优势抗原,在活疫苗或灭活全细胞疫苗中比其他两种蛋白能刺激宿主产生更强的抗体反应。这些结果不仅扩展了渗透压和铁响应蛋白的知识,而且为鉴定适合疫苗开发的保护性蛋白提供了有价值的策略。