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Fish Shellfish Immunol. 2015 Nov;47(1):239-44. doi: 10.1016/j.fsi.2015.09.016. Epub 2015 Sep 9.

A synthetic peptide derived from the D1 domain of flagellin induced the expression of proinflammatory cytokines in fish macrophages.

Author information

1
Grupo de Marcadores Inmunológicos, Laboratorio de Genética e Inmunología Molecular, Instituto de Biología, Pontificia Universidad Católica de Valparaíso, Valparaíso, Chile; Laboratorio de Biotecnología y Patología Acuática, Instituto de Patología Animal, Universidad Austral de Chile, Valdivia, Chile; Centro FONDAP, Interdisciplinary Center for Aquaculture Research (INCAR), Chile. Electronic address: rgstegmaier@gmail.com.
2
Núcleo de Biotecnología Curauma (NBC), Pontificia Universidad Católica de Valparaíso, Valparaíso, Chile.
3
Institute for Research in Biomedicine, and CIBER-BBN, Networking Centre on Bioengineering, Biomaterials and Nanomedicine, Barcelona Science Park, Barcelona, Spain; Department of Organic Chemistry, University of Barcelona, Barcelona, Spain; School of Chemistry, Yachay Tech, Yachay City of Knowledge, 100199, Urcuqui, Ecuador.
4
Instituto de Bioquímica y Microbiología, Universidad Austral de Chile, Valdivia, Chile; Escuela de Tecnología Médica, Universidad Santo Tomas, Valdivia, Chile.
5
Laboratorio de Biotecnología y Patología Acuática, Instituto de Patología Animal, Universidad Austral de Chile, Valdivia, Chile; Centro FONDAP, Interdisciplinary Center for Aquaculture Research (INCAR), Chile.
6
Department of Cell Biology and Histology, Faculty of Biology, University of Murcia, IMIB-Arrixaca, 30100, Murcia, Spain.
7
Grupo de Marcadores Inmunológicos, Laboratorio de Genética e Inmunología Molecular, Instituto de Biología, Pontificia Universidad Católica de Valparaíso, Valparaíso, Chile. Electronic address: lmercado@ucv.cl.

Abstract

Flagellin is the main protein component of flagellum in Gram negative and positive bacteria, and it is also the ligand that activates the Toll-like receptor 5 (TLR5) in fish and mammals. In higher vertebrates, flagellin induces the activation of the membrane-bound TLR5 (TLR5M), which promotes the expression of proinflammatory cytokines and chemokines, and other immunological functions. We have previously reported that recombinant flagellin from Vibrio anguillarum and its ND1 domain are able to upregulate the expression of genes encoding major the proinflammatory mediators in gilthead seabream and rainbow trout macrophages. Considering the key role of D1 domain of flagellin for binding to TLR5M and its immunostimulatory activity, we designed and chemically synthesized a peptide derived of this region. The effects of the synthetic peptide were evaluated in vitro using head kidney macrophages from gilthead seabream (Sparus aurata L., Perciformes, Sparidae) and rainbow trout (Oncorhynchus mykiss W., Salmoniformes, Salmonidae). In both species the expression of genes encoding the proinflammatory cytokines interleukin-1β (IL-1β) and tumour necrosis factor-α (TNF-α), and the chemokine IL-8, was induced upon stimulation of macrophages with the D1 domain synthetic peptide. IL-1β and IL-8 were the most upregulated genes and to a lesser extent TNF-α. Interestingly, however, the induction activity of the synthetic peptide was higher in gilthead seabream than in rainbow trout macrophages. The results were confirmed at the protein levels for IL-8. Collectively, these results suggest that synthetic peptide derived from flagelling could be a promising approach for the immunostimulation and vaccination of farmed fish.

KEYWORDS:

Cytokines; Fish immunology; Flagellin; Immunomodulatory peptide; TLR5

PMID:
26363237
DOI:
10.1016/j.fsi.2015.09.016
[Indexed for MEDLINE]

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