Identification of four type I IFNs from Japanese eel with differential expression properties and Mx promoter inducibility

Type I IFNs are a family of cytokines with antiviral, anti-proliferative and immune-modulatory functions. In this study, four type I IFNs (termed AjIFN1-4) have been cloned from the Japanese eel, Anguilla japonica. The open reading frames of AjIFN1-4 are 552, 534, 546 and 561 bp in length, encoding 183, 177, 181, and 186 amino acids (aa), respectively. Sequence comparison and phylogenetic analysis results revealed that AjIFN1 and AjIFN2 belong to group one (2C-containing) IFNs, while AjIFN3 and AjIFN4 belong to group two (4C-containing) IFNs. Syntenic comparison showed that chromosome block duplication and rearrangement events might have occurred at IFN loci in different teleost lineages. Expression analysis revealed the rapid induction of AjIFNl and AjIFN2 in response to poly I:C stimulation, while AjIFN3 and AjIFN4 were predominantly expressed at later time points. Two Mx promoter reporter assays were conducted to assess the Mx-inducing capability of AjIFN1-4. It is shown that the overexpression of AjIFN1-4 all promoted the luciferase activity of MxB reporter, but the activity of MxC reporter increased only in cells transfected with AjIFN1. Collectively, it is suggested that teleost IFNs were evolved independently in different lineages of fish and may function differently in teleost antiviral immunity.