Molecular characterizations, immune modulation, and antibacterial activity of interleukin-17A/F1a and interleukin-17A/F1b in common carp Cyprinus carpio

Abstract

Interleukin-17 (IL-17), as a pro-inflammatory cytokine family, mediates different pro-inflammatory mediators in various cell types (e.g., epithelial cells, macrophages, endothelial cells, and fibroblasts), which play an important role in defending against pathogens. The IL-17A/F1 genes have recently been reported in fish. However, the functions of these genes are still unclear. In this study, we identified two duplicated IL-17A/F1 genes in common carp (Cyprinus carpio L.), namely, CcIL-17A/F1a and CcIL-17A/F1b. Sequence analysis showed that CcIL-17A/F1a and CcIL-17A/F1b proteins had four conserved cysteine residues, which could form two intra-chain disulfide bridges. Homology comparison displayed that the deduced amino acid sequences of CcIL-17A/F1a and CcIL-17A/F1b shared 31.1%–77.3% and 32.5%–75.7% of sequence similarity to IL-17A/F1 homologues from other fish species, respectively. The mRNA expression levels of CcIL-17A/F1a and CcIL-17A/F1b were obviously increased in gill and head–kidney of fish challenged with A. hydrophila. The recombinant protein rCcIL-17A/F1a and rCcIL-17A/F1b could enhance the expression levels of pro-inflammatory cytokines (IL-1β, IL-6, IFN-γ, and TNF-α) and chemokines (CXCL8 and CXCL20). The 3 × Flag eukaryotic expression vectors to express protein rCcIL-17A/F1a (or rCcIL-17A/F1b) were constructed and intramuscularly injected in common carp. The rCcIL-17A/F1a (or rCcIL-17A/F1b) could be successfully expressed in vivo. Four immune-related genes, namely, CD4, CD8, TNF-α, and IgM, were also significantly induced to be expressed at higher mRNA levels compared with the control. The pretreatment with CcIL-17A/F1a or CcIL-17A/F1b could markedly increase the survival rate of common carp challenged with A. hydrophila. Our results demonstrated that CcIL-17A/F1a or CcIL-17A/F1b plays an important role in immune responses and immune defense against bacteria. CcIL-17A/F1a or CcIL-17A/F1b could also be potentially used as an immunopotentiator to prevent diseases in common carp.