Interleukin (IL)-22 in common carp (Cyprinus carpio L.): Immune modulation, antibacterial defense, and activation of the JAK-STAT signaling pathway

Abstract

Interleukin (IL)-22 is an IL-10 family cytokine secreted by CD4+ T cells and plays an important role in regulating inflammation and infection elimination. IL-22 homologues have been reported in the teleost, but the functions of IL-22 are still unclear. In this study, we identified two duplicated IL-22 genes in common carp (Cyprinus carpio L.), termed Cc_IL-22A and Cc_IL-22B. Sequence analysis showed that Cc_IL-22A and Cc_IL-22B had four conserved cysteine residues, which could form two intra-chain disulfide bridges. The Cc_IL-22A and Cc_IL-22B were constitutively expressed in various tissues, with the highest expression in the gill. The mRNA expression levels of Cc_IL-22A and Cc_IL-22B were significantly up-regulated in the gill, intestine, head kidney, and spleen of common carp challenged with Aeromonas. hydrophila. In vivo study showed that the expression levels of pro-inflammatory cytokines were significantly up-regulated in the head kidney and spleen when Cc_IL-22A or Cc_IL-22B were over-expressed. Furthermore, the over-expression of Cc_IL-22A and Cc_IL-22B indicated a protective effect on tissues, with only lymphocytic infiltration observed in comparison to the control and pcN3 groups, without obvious change in tissue morphology. Similar stimulatory effects of rIL-22A and rIL-22B were observed in vitro. When HKLs were stimulated with rIL-22A or rIL-22B, the expression levels of critical signaling molecules in the Janus kinase (JAK) and signal transducer and activator of transcription (STAT) pathway were significantly induced, including JAK1, JAK3, STAT1, and STAT3, as well as pro-inflammatory cytokines (IL-1β and TNF-α). Together, these results suggest that Cc_IL-22A and Cc_IL-22B may regulate inflammatory responses through the JAK-STAT signaling pathway and have a significant impact on the immune defense of common carp against bacterial infection. Therefore, our study provides a new perspective on the functions of Cc_IL-22A and Cc_IL-22B in the immune defense mechanism of fish.