Identification of Omp38 by immunoproteomic analysis and evaluation as a potential vaccine antigen against Aeromonas hydrophila in Chinese breams
Identification of Omp38 by immunoproteomic analysis and evaluation as a potential vaccine antigen against Aeromonas hydrophila in Chinese breams
复制标题
免疫蛋白质组学鉴定 Omp38 及其作为中华鳊鱼嗜水气单胞菌潜在疫苗抗原的评价
DOI：
10.1016/j.fsi.2012.10.003
复制
发表时间：
2013-01-01
影响因子：
4.7
通讯作者：
Lu, Chengping
中科院分区：
文献类型：
--
作者：
Wang, Na;Yang, Zhao;Lu, Chengping
嗜水气单胞菌是一种在水生环境中引起全身性感染的鱼类病原体，确定其抗原蛋白对开发疫苗以减少世界范围内水产养殖的经济损失具有重要意义。本研究采用免疫蛋白质组学方法，利用中华鲷鱼恢复期血清，鉴定了中国J-1疫苗株的免疫原性外膜蛋白（OMPs）。通过二维（2-D）电泳和基质辅助激光解吸电离飞行时间质谱（MALDI-TOF-TOF-MS）鉴定了7种独特的免疫原性蛋白。表达了一种感兴趣的蛋白Omp38，并对其免疫原性和保护作用进行了评价。两组经灭活疫苗和重组Omp38蛋白免疫的鱼接种后血清IgM抗体水平均显著高于注射PBS缓冲液的鱼。免疫组头肾淋巴细胞超氧化物歧化酶（SOD）活性、溶菌酶（LSZ）活性和吞噬活性均显著高于对照组。接种灭活疫苗和重组Omp38蛋白的鱼在免疫后45天对活的嗜水单胞菌攻击产生了保护性反应，这表明接种疫苗的鱼的存活率高于对照组，接种疫苗的鱼的组织学改变减少。此外，Omp38组的保护作用优于灭活疫苗组。这些结果表明，重组Omp38蛋白可以有效地刺激特异性和非特异性免疫反应，并对嗜水单胞菌感染具有保护作用。因此，Omp38可能作为一种潜在的候选疫苗来对抗嗜水单胞菌感染。(c) 2012 Elsevier Ltd.版权所有。
Aeromonas hydrophila is a fish pathogen causing systemic infections in aquatic environments, and determining its antigenic proteins is important for vaccine development to reduce economic losses in aquaculture worldwide. Here, an immunoproteomic approach was used to identify immunogenic outer membrane proteins (OMPs) of the Chinese vaccine strain J-1 using convalescent sera from Chinese breams. Seven unique immunogenic proteins were identified by two-dimensional (2-D) electrophoresis and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-TOF-MS). One protein of interest, Omp38, was expressed, and its immunogenicity and protective efficacy were evaluated in Chinese breams. The two groups of fish immunized with the inactivated vaccine and recombinant Omp38 protein showed significant serum IgM antibody levels after vaccination, compared with the fish injected with PBS buffer. In addition, the superoxide dismutase (SOD) activity, lysozyme (LSZ) activity and phagocytosis activity of head kidney lymphocytes of immunized groups were significantly higher than those of the control. The fish receiving inactivated vaccine and recombinant Omp38 protein developed a protective response to a live A. hydrophila challenge 45 days post-immunization, as demonstrated by increased survival of vaccinated fish over the control and by decreased histological alterations in vaccinated fish. Furthermore, protective effect was better in Omp38 group than in the inactivated vaccine group. These results suggest that the recombinant Omp38 protein could effectively stimulate both specific and non-specific immune responses and protect against A. hydrophila infection. Therefore, Omp38 may be developed as a potential vaccine candidate against A. hydrophila infection. (c) 2012 Elsevier Ltd. All rights reserved.