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首页> 外文期刊>Fish & Shellfish Immunology >Edwardsiella tarda mutant disrupted in type III secretion system and chorismic acid synthesis and cured of a plasmid as a live attenuated vaccine in turbot
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Edwardsiella tarda mutant disrupted in type III secretion system and chorismic acid synthesis and cured of a plasmid as a live attenuated vaccine in turbot

机译：Edwardsiella tarda突变体破坏了III型分泌系统和鸟叉酸的合成，并固化了一种质粒，作为大菱turbo中的减毒活疫苗

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Edwardsiella tarda is an intractable Gram-negative pathogen in many fish species to cause edwardsiellosis. Its infection leads to extensive losses in a diverse array of commercially important fish. The type III secretion system (T3SS) has been considered as one of the major virulence factors and plays important roles in its intracellular lifestyle. In this study, an E. tarda EIB202 mutant WED with deletions in the T3SS genes for EseB, EseC, EseD and EscA, along with the aroC gene for the biosynthesis of chorismic acid, as well as the curing of endogenous plasmid pEIB202 was constructed by allelic exchange strategy. Compared to the wild-type EIB202 which was highly virulent towards turbot (Scophthamus maximus) via intraperitoneal (i.p.), intramuscular (i.m.) injection or immersion and caused systemic infection in turbot as well as the unexpected red mouth symptom when immersion challenged, WED was highly attenuated when inoculated into turbot via i.m., i.p. and immersion routes, and exhibited significantly impaired capacity to survive in fish tissues. WED showed 5700-fold higher 50% lethal dose (LD50) than that of the wild type when i.m. or i.p. challenged. Inoculation with WED by i.p. or immersion injection routes elicited significant protection against the challenge of the wild-type E. tarda after 5 weeks of vaccination. The vaccinated fish produced low while significant level of specific antibody and showed increased expression of immune-related factors including IL-1 beta, IFN-gamma, MHC II, MHC-I and CD8, indicating that WED possesses significant immunoprotective potential. Furthermore, our data indicated that a single dose of i.p. and immersion vaccination with WED could produce significant protection as long as 12 and 6 months, respectively. These results demonstrated the feasibility of WED as a live attenuated vaccine in turbot against edwardsiellosis by immersion or i.p. injection routes. (C) 2013 Elsevier Ltd. All rights reserved.
机译：泰氏爱德华氏菌是许多鱼类中难治的革兰氏阴性病原体，可引起埃德氏病。它的感染导致各种重要商业鱼类的大量损失。 III型分泌系统（T3SS）被认为是主要的毒力因子之一，并在其细胞内生活方式中发挥重要作用。在这项研究中，构建了一个E.tarda EIB202突变体WED，该突变体在EseB，EseC，EseD和EscA的T3SS基因中缺失，以及用于鸟苷酸生物合成的aroC基因以及内源质粒pEIB202的固化等位基因交换策略。与野生型EIB202相比，野生型EIB202通过腹膜内（ip），肌内（im）注射或浸入对大菱t（Scophthamus maximus）具有极强的毒力，在浸没攻击时会导致大菱system中的全身感染以及意外的红口症状。通过im，ip接种到大菱t中时高度衰减和浸没途径，并且在鱼类组织中的生存能力显着受损。 WED的i.m.值显示其50％致死剂量（LD50）比野生型高5700倍。或i.p.挑战。 i.p.接种WED接种5周后，注射或浸泡注射途径可显着保护其免受野生型塔氏大肠杆菌的攻击。疫苗接种的鱼产生的抗体水平低而显着，但免疫相关因子（包括IL-1β，IFN-γ，MHC II，MHC-1和CD8）的表达增加，表明WED具有显着的免疫保护潜力。此外，我们的数据表明腹膜内注射单剂量。 WED和浸入式疫苗接种分别可分别提供长达12个月和6个月的有效保护。这些结果证明了WED作为通过浸没或腹膜内淋巴结菌防治大肠埃希菌病的减毒活疫苗的可行性。注射途径。 （C）2013 Elsevier Ltd.保留所有权利。

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