Identification, expression and pro-inflammatory effect of interleukin-17 N in common carp (Cyprinus carpio L.)

Abstract

Two interleukin (IL)-17 N genes (CcIL-17Na and b) present on different linkage groups were identified in the common carp (Cyprinus carpio) genome and confirmed by polymerase chain reaction (PCR) and real time (RT)-PCR in this experiment. Synteny analysis revealed that IL-17 N is transcribed by the complement sequence of TOP3B's intron 2. It is flanked by SDF2L and PPM1F in all fish studied to date, except fugu (Takifugu rubripes). The open reading frames of the two CcIL-17Ns are 411 base pairs long and encode 136 amino acids. The amino acid identity/similarity between CcIL-17Na and b is 91.2%/97.1%. The CcIL-17Ns share identity (46.8–90.4%) with their orthologs from other teleosts. Identities/similarities to other members of the IL-17 family in common carp were low at 21.4–30.2%/31.4–51.4%. In the phylogenetic tree, IL-17Ns from spotted gar (Lepisosteus oculatus, the ancestor of teleosts) and coelacanth (Latimeria chalumnae, the ancestor of tetrapods) were grouped within the same branch with a high bootstrap value of 97%, which indicates that IL-17 N is an ancient and conserved gene. Quantitative RT-PCR results showed that CcIL-17Ns were most highly expressed in the brain of healthy individuals. The expression in brain was significantly induced at 6 h post Aeromonas hydrophila infection; at 1 day post infection, expression in liver, muscle, skin, spleen, and head kidney was up-regulated. In addition, the upregulated expression of proinflammatory cytokines IL-1β, IFN-γ, IL-6, chemokine CCL20, NF - κ B and TRAF6 in kidney tissue by ccIL-17 N recombinant protein also indicate that IL-17 N can promote inflammation through NF-κB pathway and induce the expression of chemokines and inflammatory factors.