Molecular characteristics of interleukin (IL)-17A/F3 and its immune response on the pathogen and functional regulation on cytokines in common carp Cyprinus carpio L.

Abstract

Fish interleukin (IL)-17A/F is homologous with mammalian IL-17A and IL-17F, which plays a key role in regulating inflammatory responses and autoimmune diseases. In fish, IL-17A/F1, 2, and 3 have been identified and described. However, IL-17A/F3 has received little attention in fish. In this study, a homolog of IL-17A/F3 was identified in common carp (Cyprinus carpio L.), which was termed as Cc_IL-17A/F3. The deduced amino acid sequence of Cc_IL-17A/F3 has four conserved cysteine residues, which could form two intrachain disulfide bonds. Homology comparison showed that the Cc_IL-17A/F3 was in the range of 31.7–71.9% of sequence similarity with these of other fishes. The Cc_IL-17A/F3 gene was constitutively expressed in various tissues, with higher expression levels in the skin and gills. After common carp were infected by Aeromonas. hydrophila, the mRNA expression levels of Cc_IL-17A/F3 were significantly up-regulated in the spleen, head kidney, gills, and intestine. Based on the indirect immunofluorescence assay, Cc_IL-17A/F3 proteins were found to be obviously increased in the intestine and spleen upon A. hydrophila infection at 24 h post-infection. The recombinant protein rCc_IL-17A/F3 could enhance the gene expression levels of pro-inflammatory cytokines (IL-1β, IL-6, IFN-γ, and TNF-α) as well as chemokines (CXCL8 and CXCL20) in primary head kidney leukocytes. In vivo and in vitro experiments have similar stimulatory effects. When Cc_IL-17A/F3 was overexpressed in common carp, the expressions of pro-inflammatory cytokines and chemokines were significantly up-regulated in head kidney and spleen. In summary, the results derived from the present study suggested that the Cc_IL-17A/F3 plays an important role in defending against bacterial infections, and probably participates in mucosal immunity of the host.