Characterisation and expression analysis of the interleukin genes, IL-1beta, IL-8 and IL-10, in Atlantic cod (Gadus morhua L.).
The mammalian interleukins IL-1beta and IL-8 are important pro-inflammatory cytokines often used as markers of an activated inflammatory response, while IL-10 is regarded as an anti-inflammatory cytokine and plays a crucial role in the regulation of inflammation. Few cytokines from gadoid fish have been described, and herein the sequence and expression of these interleukin genes are studied in Atlantic cod (Gadus morhua L.). IL-1beta, IL-8 and IL-10 from cod show similarities in gene organisation and predicted protein sequences, compared to counterpart genes in other teleosts. Gene expression was studied using quantitative real time PCR in response to several treatments both in vitro and in vivo. In adherent head kidney cells, infectious pancreatic necrosis virus (IPNV) and lipopolysaccharide (LPS) significantly stimulated gene expression of IL-1beta. The expression of IL-1beta was not increased after treatment with a viral imitator (poly I:C), and neither IL-8 nor IL-10 responded to any of these agents in vitro. However, in vivo administrated poly I:C and formalin-killed Vibrio anguillarum (In-V.ang) induced interleukin expression, varying in intensity between different organs. IL-1beta and IL-10 gene expression profiles showed an opposite induction pattern in the in vivo experiments. Injection of In-V.ang highly induced IL-1beta expression, while a low induction was evident for IL-10, whereas the opposite was observed after injection of poly I:C. This pattern was particularly marked in spleen, where also IL-8 followed the expression pattern of IL-1beta. The opposite expression profiles indicate a suppressive role for IL-10 on the transcription of IL-1beta, and to a lesser extent on IL-8 transcription. Along with the identification of important promoter regulatory motives, these results provide tools for studying inflammatory responses in cod.
Seppola
Marit
M
Fiskeriforskning, Norwegian Institute of Fisheries and Aquaculture Research, N-9291 Tromsø, Norway. marit.seppola@fiskeriforskning.no
Larsen
Atle Noralf
AN
Steiro
Kari
K
Robertsen
Børre
B
Jensen
Ingvill
I
D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D048389:Gadus morhua / Q000276:immunology* / Q000821:virology; D020869:Gene Expression Profiling; D018061:Infectious pancreatic necrosis virus / Q000502:physiology; D016753:Interleukin-10 / Q000096:biosynthesis* / Q000235:genetics; D053583:Interleukin-1beta / Q000096:biosynthesis* / Q000235:genetics; D016209:Interleukin-8 / Q000096:biosynthesis* / Q000235:genetics; D008070:Lipopolysaccharides / Q000494:pharmacology; D008969:Molecular Sequence Data; D010802:Phylogeny; D011070:Poly I-C / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D014733:Vibrio / Q000502:physiology
D053583:Interleukin-1beta; D016209:Interleukin-8; D008070:Lipopolysaccharides; D016753:Interleukin-10; D011070:Poly I-C