Characterisation and expression analysis of the Atlantic halibut (Hippoglossus hippoglossus L.) cytokines: IL-1b, IL-6, IL-11, IL-12b and IFNc
2011
Øvergård, Aina-Cathrine | Nepstad, Ina | Nerland, Audun Helge | Patel, Sonal
Genes encoding the five Atlantic halibut (Hippoglossushippoglossus L.) cytokines; interleukin (IL)-1b,IL-6, IL-11b, IL-12bc, and interferon (IFN) c, were clonedand characterised at a molecular level. The genomic organisationof the halibut cytokine genes was similar to thatseen in mammals and/or other fish species. Several mRNAinstability motifs were found within the 30-untranslatedregion (UTR) of all cytokine cDNA sequences. The putativecytokine protein sequences showed a low sequence identitywith the corresponding homologues in mammals, avian andother fish species. Nevertheless, important structural featureswere presumably conserved such as the presence, orabsence in the case of IL-1b, of a signal peptide, secondarystructure and family signature motifs. The relative expressionpattern of the cytokine genes was analyzed in severalhalibut organs, revealing a constitutive expression in bothlymphoid and non-lymphoid organs. Interestingly, the gillsshowed a relatively high expression of IL-1b, IL-12bc andIFNc. The real time RT-PCR data also showed that themRNA level of IL-1b, IL-6, IL-12bc and IFNc was high inthe thymus, while IL-11b was relatively highly expressed inthe posterior kidney and posterior gut. Moreover, the halibutbrain showed a relatively high level of IL-6 transcripts.Anterior kidney leucocytes in vitro stimulated with imiquimodshowed a significant increase in mRNA level of thefive halibut cytokine genes. The sequence and characterisationdata presented here will be useful for furtherinvestigation of both innate and adaptive immune responsesin halibut, and be helpful in the design of vaccines for thecontrol of various infectious diseases.
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