Nile tilapia TLR3 recruits MyD88 and TRIF as adaptors and is involved in the NF-κB pathway in the immune response

TLR3 plays a crucial role in innate immunity. In the present study, OnTLR3 was identified in the Nile tilapia Oreochromis niloticus, with a conserved LRR domain and a C-terminal TIR domain. OnTLR3 was broadly expressed in all tissues tested, with the highest expression levels in the blood and the lowest in the kidney. TLR3 mRNA could be detected from pharyngula (2.5 dpf) to late larva (8.5 dpf) during embryonic and larval development. Moreover, the expression level of OnTLR3 was clearly altered in all five tissues after Streptococcus agalactiae infection in vivo and could be induced by LPS, poly(I:C), S. agalactiae WC1535 and △CPS in Nile tilapia macrophages. When OnTLR3 was overexpressed in 293 T cells, it was distributed in the cytoplasm and could significantly increase NF-κB activation. The pulldown assays showed that OnTLR3 interacted with both OnMyD88 and OnTRIF. The binding assays revealed the specificity of OnTLR3 for pathogen-associated molecular patterns (PAMPs) and bacteria that included S. agalactiae, Aeromonas hydrophila and poly(I:C), LPS and PGN. Taken together, these findings suggest that OnTLR3, as a pattern recognition receptor (PRR), might play an important role in the immune response to pathogen invasion. •TLR3 gene was identified in Nile tilapia and was upregulated after S. agalactiae challenge.•TLR3 induced by LPS, Poly I: C, wild and mutant type S. agalactiae in in vitro.•TLR3 interact with MyD88 and TRIF.•The overexpression of TLR3 significantly induced NF-κB activity in cell line.•Binding assays revealed the specificity of OnTLR3 for pathogen-associated molecular patterns (PAMPs).