Establishmentof fish disease diagnosis using cytokinenetwork

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The multiplex RT-PCR assay targeting 19 cytokine genes was constructed and the expression of cytokine genes in Fuguin response to immunostimulants or pathogenic bacteria infection was analyzed using this method. As results, increased expressionof pro-inflammatory cytokines was observed in all the experiments. Moreover, the increase in I-IFN gene expression was confirmed only by stimulation with viral mimic polyI:C. Next, production of IL-10, IL-17A/F1, IFN-y and IL-4/13 was analyzed in the kidney of Fuguinfected with pathogenic bacteria or parasite. The production of all cytokines analyzed was increased by the bacterial infection. Although similar production pattern was confirmed in parasite infection also, especially a conspicuous increase in IL-4/13 level was recorded. Next, the detection method for intracellular cytokine was established and it enabled us to detect IFN-y producing cells. Using this method, we found the increased number of IFN-y producing cells in peripheral blood leucocytes in the stimulation with bacterial component.