Design and functional characterization of Salmo salar TLR5 agonist peptides derived from high mobility group B1 acidic tail

• Published in: Fish & shellfish immunology 2024-03, Vol.146, p.109373-109373, Article 109373
• Main Authors: Vásquez-Suárez, Aleikar, Muñoz-Flores, Carolina, Ortega, Leonardo, Roa, Francisco, Castillo, Carolina, Romero, Alex, Parra, Natalie, Sandoval, Felipe, Macaya, Luis, González-Chavarría, Iván, Astuya, Allisson, Starck, María Francisca, Villegas, Milton F., Agurto, Niza, Montesino, Raquel, Sánchez, Oliberto, Valenzuela, Ariel, Toledo, Jorge R., Acosta, Jannel
• Format: Article
• Language: English
• Subjects: HMGB1; Immune response; Immunostimulant; Piscirickettsia salmonis; Salmonids; TLR5
• ISSN: 1050-4648; 1095-9947
• DOI: 10.1016/j.fsi.2024.109373

Toll-like receptor 5 (TLR5) responds to the monomeric form of flagellin and induces the MyD88-depending signaling pathway, activating proinflammatory transcription factors such as NF-κB and the consequent induction of cytokines. On the other hand, HMGB1 is a highly conserved non-histone chromosomal protein shown to interact with and activate TLR5. The present work aimed to design and characterize TLR5 agonist peptides derived from the acidic tail of Salmo salar HMGB1 based on the structural knowledge of the TLR5 surface using global molecular docking platforms. Peptide binding poses complexed on TLR5 ectodomain model from each algorithm were filtrated based on docking scoring functions and predicted theoretical binding affinity of the complex. Circular dichroism spectra were recorded for each peptide selected for synthesis. Only intrinsically disordered peptides (6W, 11W, and SsOri) were selected for experimental functional assay. The functional characterization of the peptides was performed by NF-κB activation assays, RT-qPCR gene expression assays, and Piscirickettsia salmonis challenge in SHK-1 cells. The 6W and 11W peptides increased the nuclear translation of p65 and phosphorylation. In addition, the peptides induced the expression of genes related to the TLR5 pathway activation, pro- and anti-inflammatory response, and differentiation and activation of T lymphocytes towards phenotypes such as TH1, TH17, and TH2. Finally, it was shown that the 11W peptide protects immune cells against infection with P. salmonis bacteria. Overall, the results indicate the usefulness of novel peptides as potential immunostimulants in salmonids. •TLR5 agonist peptides designed from the acidic tail of HMGB1 activate the NF-κB pathway.•Both 6W and 11W peptides induce up-regulation of immune-related genes in vitro and in vivo in salmonids.•The 11W peptide prevents the cytotoxicity induced by P. salmonis infection in the SHK-1 cells.•The novel peptides are potential candidates for establishing of immunostimulant formulations in salmonids.