Identification of interleukin-22 in gadoids and examination of its expression level in vaccinated fish

Abstract

This paper reports the cloning and sequencing of interleukin (IL)-22 in two gadoid fish, cod (Gadus morhua) and haddock (Melanogrammus aeglefinus). The complete transcript of this gene was 1002 and 1154bp respectively, of which 492bp was the open reading frame (ORF) in both genes. High amino acid identity (88.3%) was found between these genes but was less than 50% aa identity to other known genes. The gene organisation of haddock IL-22 consisted of five exons and four introns, as with other IL-10 family members. Expression studies showed that IL-22 is constitutively expressed in gill, with low level expression also observed in gut, gonad and head kidney. In a vaccination experiment, haddock were injected intraperitoneally with formalin-killed Vibrio anguillarum or with PBS, and 2 months later challenged by immersion in 107cfu/ml V. anguillarum for 30min. Head kidney and gill samples were collected prior to challenge and 24, 48 and 72h post-challenge (hpc) for Real-time PCR analysis of IL-22 expression. No significant changes in IL-22 expression were observed in head kidney tissue but vaccinated fish showed a significantly increased expression of IL-22 24hpc in gill and no mortalities were seen in these fish. In contrast, control fish, which started to succumb to the disease from 72hpc, showed no significant increase in gill expression after challenge.