A protective immune response is generated in rainbow trout by an OmpH-like surface antigen (P18) of Flavobacterium psychrophilum
A protective immune response is generated in rainbow trout by an OmpH-like surface antigen (P18) of Flavobacterium psychrophilum
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DOI：
10.1128/aem.00279-06
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发表时间：
2006-07-01
影响因子：
4.4
通讯作者：
Le Henaff, Michel
中科院分区：
文献类型：
--
作者：
Dumetz, Fabien;Duchaud, Eric;Le Henaff, Michel
调查的表面特征的黄杆菌psychrophilum，一种重要的鱼类病原体，帮助我们确定一个表面蛋白P18。在目前的研究中，我们开发了一种简单而有效的程序，通过两步法纯化这种蛋白质。首先，P18选择性地释放从黄细菌的热HEPES处理的细胞，然后进行阴离子交换高效液相色谱。使用从头测序从纯化的P18产生片段化肽谱。将获得的两个肽段序列与F. psychrophilum（INRA，茹伊-恩-乔萨，法国）鉴定了一个编码166个氨基酸的OmpH样蛋白的基因，该蛋白很可能经历23个残基信号肽的N-末端切割。OmpH样蛋白对蛋白酶K处理的敏感性和抗OmpH样蛋白抗体的抑菌/杀菌活性表明该蛋白实际上暴露在F.嗜冷菌免疫接种试验表明OmpH样蛋白可诱导产生高滴度的抗OmpH样蛋白抗体，具有保护性。综上所述，这些结果表明，由F.嗜冷菌可作为一种有希望的候选抗原用于未来的疫苗开发。
Investigations of the surface characteristics of Flavobacterium psychrophilum, an important pathogen of fish, assisted us in identifying a surface protein termed P18. In the current study, we developed a simple and efficient procedure for the purification of this protein by a two-step method. First, P18 was selectively released from flavobacteria by a heat-HEPES treatment of the cells and then subjected to anion-exchange high-performance liquid chromatography. De novo sequencing was used to generate a fragmented peptide spectrum from purified P18. Comparison of two obtained peptide sequences with a partial genome sequence of F. psychrophilum (INRA, Jouy-en-josas, France) identified one gene encoding a 166-amino-acid OmpH-like protein that mostly likely undergoes N-terminal cleavage of the 23-residue signal peptide. The susceptibility of the OmpH-like protein to proteinase K treatment and the bacteriostatic/bactericidal activities of anti-OmpH-like protein antibodies indicated that this protein is actually exposed on the surface of F. psychrophilum. Vaccination trials showed that the OmpH-like protein can induce a high titer of anti-OmpH-like protein antibodies which are protective. Taken together, these results suggest that this surface protein produced by F. psychrophilum could be used in future vaccine development as a promising candidate antigen.