TY - JOUR
T1 - A synthetic peptide derived from the D1 domain of flagellin induced the expression of proinflammatory cytokines in fish macrophages
AU - González-Stegmaier, Roxana
AU - Guzmán, Fanny
AU - Albericio, Fernando
AU - Villarroel-Espíndola, Franz
AU - Romero, Alex
AU - Mulero, Victoriano
AU - Mercado, Luis
N1 - Funding Information:
We thank I. Fuentes for excellent technical assistance. This work was supported by the Seventh Framework Programme of the European Union (grant agreement TARGETFISH 311993 to VM) and Dirección de Investigación, Pontificia Universidad Católica de Valparaíso, Chile (grant ATS13.11 to RGS); Advanced Human Capital Formation of CONICYT; BECA CHILE Scholarship for doctoral studies (to RGS) and FONDAP 15110027; Fondo Nacional de Desarrollo en Ciencia y Tecnología FONDECYT 3130449 (to FVE).
Publisher Copyright:
© 2015 Elsevier Ltd.
PY - 2015/11/1
Y1 - 2015/11/1
N2 - 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.
AB - 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.
KW - Cytokines
KW - Fish immunology
KW - Flagellin
KW - Immunomodulatory peptide
KW - TLR5
UR - http://www.scopus.com/inward/record.url?scp=84941961601&partnerID=8YFLogxK
U2 - 10.1016/j.fsi.2015.09.016
DO - 10.1016/j.fsi.2015.09.016
M3 - Article
C2 - 26363237
AN - SCOPUS:84941961601
VL - 47
SP - 239
EP - 244
JO - Fish and Shellfish Immunology
JF - Fish and Shellfish Immunology
SN - 1050-4648
IS - 1
ER -