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大黄鱼弧菌外膜蛋白 U 亚单位疫苗的免疫原性研究

Immunogenicity study of OmpU subunit vaccine against Vibrio mimicus in yellow catfish, Pelteobagrus fulvidraco.

机构信息

State Key Laboratory of Agricultural Microbiology, College of Fisheries, Huazhong Agricultural University, Wuhan, Hubei, China; Engineering Research Center of Green Development for Conventional Aquatic Biological Industry in the Yangtze River Economic Belt, Ministry of Education, Wuhan, Hubei, China.

Agriculture Ministry Key Laboratory of Healthy Freshwater Aquaculture, Key Laboratory of Fish Health and Nutrition of Zhejiang Province, Zhejiang Institute of Freshwater Fisheries, Huzhou, Zhejiang, China.

出版信息

Fish Shellfish Immunol. 2021 Jan;108:80-85. doi: 10.1016/j.fsi.2020.11.030. Epub 2020 Dec 4.

Abstract

The outer membrane protein U (OmpU) is a conserved outer membrane protein in a variety of pathogenic Vibrio species and has been considered as a vital protective antigen for vaccine development. Vibrio mimicus (V. mimicus) is the pathogen causing ascites disease in aquatic animals. In this study, the prokaryotically expressed and purified His-tagged OmpU of V. mimicus (His-OmpU) was used as a subunit vaccine. The formalin inactivated V. mimicus, purified His tag (His-tag), and PBS were used as controls. The vaccinated yellow catfish were challenged with V. mimicus at 28 days post-vaccination, and the results showed that the His-OmpU and inactivated V. mimicus groups exhibited much higher survival rates than the His-tag and PBS groups. To fully understand the underlying mechanism, we detected the expression levels of several immune-related genes in the spleen of fish at 28 days post-vaccination and 24 h post-challenge. The results showed that most of the detected immune-related genes were significantly upregulated in His-OmpU and inactivated V. mimicus groups. In addition, we performed the serum bactericidal activity assay, and the results showed that the serum from His-OmpU and inactivated V. mimicus groups exhibited much stronger bactericidal activity against V. mimicus than those of His-tag and PBS groups. Finally, the serum agglutination antibody was detected, and the antibody could be detected in His-OmpU and inactivated V. mimicus groups with the antibody titers increasing along with the time post-vaccination, but not in His-tag or PBS group. Our data reveal that the recombinant OmpU elicits potent protective immune response and is an effective vaccine candidate against V. mimicus in yellow catfish.

摘要

外膜蛋白 U(OmpU)是多种致病性弧菌属物种中保守的外膜蛋白,被认为是疫苗开发的重要保护性抗原。Vibrio mimicus(V. mimicus)是引起水生动物腹水病的病原体。在本研究中,使用原核表达和纯化的 His 标签的 V. mimicus 的 OmpU(His-OmpU)作为亚单位疫苗。福尔马林灭活的 V. mimicus、纯化的 His 标签(His-tag)和 PBS 用作对照。在接种后 28 天,用 V. mimicus 攻毒接种黄颡鱼,结果表明,His-OmpU 和灭活 V. mimicus 组的存活率明显高于 His-tag 和 PBS 组。为了充分了解潜在的机制,我们在接种后 28 天和攻毒后 24 小时检测了鱼脾脏中几种免疫相关基因的表达水平。结果表明,大多数检测到的免疫相关基因在 His-OmpU 和灭活 V. mimicus 组中显著上调。此外,我们进行了血清杀菌活性测定,结果表明,His-OmpU 和灭活 V. mimicus 组的血清对 V. mimicus 的杀菌活性明显强于 His-tag 和 PBS 组。最后,检测了血清凝集抗体,结果表明,His-OmpU 和灭活 V. mimicus 组可检测到抗体,且随着接种后时间的延长,抗体滴度逐渐增加,但 His-tag 或 PBS 组则没有。我们的数据表明,重组 OmpU 引发了有效的保护性免疫应答,是黄颡鱼抗 V. mimicus 的有效疫苗候选物。

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