Li Hui, Xiong Xiao-Peng, Peng Bo, Xu Chang-Xin, Ye Ming-Zhi, Yang Tian-Ci, Wang San-Ying, Peng Xuan-Xian
Center for Proteomics, State Key Laboratory of Bio-Control, School of Life Sciences, Sun Yat-Sen University, University City, Guangzhou, PR China.
J Proteome Res. 2009 Sep;8(9):4342-9. doi: 10.1021/pr900439j.
Bacterium is still one of the major causes of life-threatening microbial infections. The most effective way to control bacterial infections is probably vaccine prevention. However, development of bacterial vaccine, especially polyvalent vaccines that could be used to fight against a variety of bacterial serotypes and species, is challenging due to the difficulty in identifying broad cross-protective antigens for different serotypes and species of pathogenic bacteria. In the present study, we developed a new approach to identify polyvalent vaccine candidates from outer membrane (OM) proteins of Vibrio alginolyticus with the ability to fight against infections caused by different genera and families of Gram-negative bacteria. This approach combined heterogeneous antiserum-based immunoproteomics with bacterial immunization challenging method. Four of the 35 protein spots resolved in a 2-DE gel of V. alginolyticus sarcosine-insoluble fraction could be recognized not only by homogeneous antiserum, but also by heterogeneous antisera obtained from other bacterial species, genera and families. The genes encoding the four OM proteins were then cloned and expressed in E. coli BL21. The expressed recombinant proteins were used as broadly cross-protective immunogens to immunize carps for investigation of their cross-protective spectra, activities and protective abilities in carps. The carps immunized with OmpA (VA0764) and Pal (VA1061) have abilities to fight against infections not only caused by V. alginolyticus, but also by Aeromonas hydrophila and Pseudomonas fluorescens. This study provides a novel approach for the identification of broadly cross-protective antigens, and possibly polyvalent vaccines against a variety of microbial infections.
细菌仍然是危及生命的微生物感染的主要原因之一。控制细菌感染最有效的方法可能是疫苗预防。然而，由于难以确定针对不同血清型和致病细菌种类的广泛交叉保护抗原，细菌疫苗的研发，尤其是可用于对抗多种细菌血清型和种类的多价疫苗，具有挑战性。在本研究中，我们开发了一种新方法，从溶藻弧菌的外膜（OM）蛋白中鉴定多价疫苗候选物，其能够对抗由不同属和科的革兰氏阴性细菌引起的感染。该方法将基于异源抗血清的免疫蛋白质组学与细菌免疫攻毒方法相结合。在溶藻弧菌肌氨酸不溶部分的二维凝胶中分离出的35个蛋白点中的4个，不仅能被同源抗血清识别，还能被从其他细菌种类、属和科获得的异源抗血清识别。然后克隆编码这四种OM蛋白的基因并在大肠杆菌BL21中表达。表达的重组蛋白用作广泛交叉保护免疫原，免疫鲤鱼以研究它们在鲤鱼中的交叉保护谱、活性和保护能力。用OmpA（VA0764）和Pal（VA1061）免疫的鲤鱼不仅有能力抵抗由溶藻弧菌引起的感染，还能抵抗嗜水气单胞菌和荧光假单胞菌引起的感染。本研究为鉴定广泛交叉保护抗原以及可能的针对多种微生物感染的多价疫苗提供了一种新方法。
