利用孢子展示技术对鱼类进行口服疫苗接种以预防弧菌病。

Oral vaccination of fish against vibriosis using spore-display technology.

机构信息

Departamento de Biologia, Faculdade de Ciências, Universidade do Porto, Porto, Portugal.

Centro Interdisciplinar de Investigação Marinha e Ambiental, Universidade do Porto (CIMAR/CIIMAR), Matosinhos, Portugal.

出版信息

Front Immunol. 2022 Oct 13;13:1012301. doi: 10.3389/fimmu.2022.1012301. eCollection 2022.

Abstract

Oral vaccines are highly demanded by the aquaculture sector, to allow mass delivery of antigens without using the expensive and labor-intensive injectable vaccines. These later require individual handling of fish, provoking stress-related mortalities. One possible strategy to create injection-free vaccine delivery vehicles is the use of bacterial spores, extremely resistant structures with wide biotechnological applications, including as probiotics, display systems, or adjuvants. Bacterial spores, in particular those of , have been shown to behave as mucosal vaccine adjuvants in mice models. However, such technology has not been extensively explored against fish bacterial disease. In this study, we used a laboratory strain of , for which a variety of genetic manipulation tools are available, to display at its spores surface either a antigenic protein, OmpK, or the green fluorescence protein, GFP. When previously vaccinated by immersion with the OmpK- carrying spores, zebrafish survival upon a bacterial challenge with and , increased up to 50 - 90% depending on the pathogen targeted. Further, we were able to detect anti-GFP-antibodies in the serum of European seabass juveniles fed diets containing the GFP-carrying spores and anti- antibodies in the serum of European seabass juveniles fed the OmpK-carrying spores containing diet. More important, seabass survival was increased from 60 to 86% when previously orally vaccinated with in-feed OmpK- carrying spores. Our results indicate that spores can effectively be used as antigen-carriers for oral vaccine delivery in fish.

摘要

口服疫苗受到水产养殖业的高度需求,可实现抗原的大规模传递,而无需使用昂贵且劳动密集型的注射用疫苗。后者需要对鱼类进行个体处理,从而引发与应激相关的死亡率。创造无注射疫苗输送载体的一种可能策略是利用细菌孢子,这种具有广泛生物技术应用的极其耐受力结构,包括作为益生菌、展示系统或佐剂。细菌孢子,特别是 的孢子,已被证明在小鼠模型中作为黏膜疫苗佐剂发挥作用。然而,这种技术尚未在鱼类细菌性疾病方面得到广泛探索。在本研究中,我们使用了一种实验室菌株 ,该菌株具有多种遗传操作工具,可在其孢子表面展示要么是一种抗原性蛋白 OmpK,要么是绿色荧光蛋白 GFP。当先前通过浸泡免疫带有 OmpK 的孢子时,斑马鱼在受到 和 细菌挑战时的存活率提高了 50-90%,具体取决于目标病原体。此外,我们能够在摄食含有 GFP 孢子的饲料的欧洲海鲈鱼幼鱼血清中检测到抗 GFP 抗体,以及在摄食含有 OmpK 孢子的饲料的欧洲海鲈鱼幼鱼血清中检测到抗 OmpK 抗体。更重要的是,当先前通过口服喂食含 OmpK 的孢子进行免疫时,海鲈鱼的存活率从 60%提高到 86%。我们的研究结果表明, 孢子可以有效地用作鱼类口服疫苗输送的抗原载体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/66cd/9608137/88fdd4c2ca20/fimmu-13-1012301-g001.jpg

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