Establishmentof fish disease diagnosis using cytokinenetwork

建立了针对19个细胞因子基因的多重RT-PCR方法，并利用该方法分析了细胞因子基因在免疫刺激剂或病原菌感染后的表达。结果，在所有实验中都观察到促炎症细胞因子的表达增加。此外，只有在病毒模拟PolyI：C刺激下，I-干扰素基因的表达才能得到证实。其次，分析了感染病原菌或寄生虫的Fuguis肾脏产生IL-10、IL-17A/F1、干扰素-γ和IL-4/13的情况。所有被分析的细胞因子的产生都因细菌感染而增加。虽然在寄生虫感染中也证实了类似的生产模式，但尤其是IL-4/13水平显著升高。接下来，建立了细胞内细胞因子的检测方法，使我们能够检测产生干扰素-γ的细胞。利用这种方法，我们发现在含有细菌成分的刺激下，外周血白细胞中产生干扰素-γ的细胞数量增加。

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.