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新型策略设计、生产和预纯化具有免疫调节活性的鲑鱼鱼鳔嵌合肽

New strategy for the design, production and pre-purification of chimeric peptide with immunomodulatory activity in Salmosalar.

机构信息

Biotechnology and Biopharmaceuticals Laboratory, Departamento de Fisiopatología, Facultad de Ciencias Biológicas, Universidad de Concepción. Víctor Lamas 1290, P.O. Box 160-C, Concepción, Chile.

Salmones Antártica S.A., Camino Los Ángeles, Santa Bárbara, Km. 12,8, BioBio, Chile.

出版信息

Fish Shellfish Immunol. 2022 Jun;125:120-127. doi: 10.1016/j.fsi.2022.04.034. Epub 2022 May 10.

Abstract

The intensive salmon farming is associated with massive outbreaks of infections. The use of antibiotics for their prevention and control is related to damage to the environment and human health. Antimicrobial peptides (AMPs) have been proposed as an alternative to the use of antibiotics for their antimicrobial and immunomodulatory activities. However, one of the main challenges for its massive clinical application is the high production cost and the complexity of chemical synthesis. Thus, recombinant DNA technology offers a more sustainable, scalable, and profitable option. In the present study, using an AMPs function prediction methodology, we designed a chimeric peptide consisting of sequences derived from cathelicidin fused with the immunomodulatory peptide derived from flagellin. The designed peptide, CATH-FLA was produced by recombinant expression using an easy pre-purification system. The chimeric peptide was able to induce IL-1β and IL-8 expression in Salmo salar head kidney leukocytes, and prevented Piscirickettsia salmonis-induced cytotoxicity in SHK-1 cells. These results suggest that pre-purification of a recombinant AMP-based chimeric peptide designed in silico allow obtaining a peptide with immunomodulatory activity in vitro. This could solve the main obstacle of AMPs for massive clinical applications.

摘要

集约化的三文鱼养殖会引发大规模的感染爆发。为了预防和控制这些感染,抗生素的使用与环境和人类健康的损害有关。抗菌肽(AMPs)因其具有抗菌和免疫调节活性而被提议作为抗生素的替代品。然而,其大规模临床应用的主要挑战之一是高生产成本和化学合成的复杂性。因此,重组 DNA 技术提供了更可持续、可扩展和更有利可图的选择。在本研究中,我们使用一种 AMPs 功能预测方法,设计了一种由抗菌肽 cathelicidin 序列与来自鞭毛蛋白的免疫调节肽融合而成的嵌合肽。设计的肽 CATH-FLA 通过使用易于预纯化系统的重组表达进行生产。该嵌合肽能够诱导大西洋三文鱼头肾白细胞表达 IL-1β 和 IL-8,并防止 Piscirickettsia salmonis 诱导的 SHK-1 细胞毒性。这些结果表明,对基于重组 AMP 的嵌合肽进行计算机设计的预纯化允许获得一种具有体外免疫调节活性的肽。这可能解决 AMPs 大规模临床应用的主要障碍。

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