The determination of innate immune responses using cytokine genes in fish

为了监测细胞因子基因在日本河豚鱼中的表达，开发了一种新的定量多重分析平台。该定制设计的多重RT-PCR测定用于分析免疫刺激条件下头肾细胞中19种细胞因子基因的表达谱，包括促炎、抗炎、T细胞增殖/分化、B细胞活化/分化、NK细胞刺激、抗病毒活性诱导和单核细胞/巨噬细胞祖细胞增殖细胞因子。在未刺激的对照和免疫刺激的细胞中的表达谱是不同的。此外，由于IL-1b、IL-6、IL-10、IL-12 p35、IL-12 p40、IL-21、TNF-α、TNF-N、I-IFN-1和IFN-γ基因的不同刺激，观察到表达谱增加。这些结果表明，细胞因子基因可以作为生物标志物来了解鱼类的先天免疫状态。

To monitor the expression of cytokine genes in Japanese pufferfish, a novel platform for quantitative multiplexed analysis was developed. This custom-designed multiplex RT-PCR assay was used to analyze the expression profiles of 19 cytokine genes, including pro-inflammatory, anti-inflammatory, T-cell proliferation/differentiation, B-cell activation/differentiation, NK cell stimulation, induction of anti-viral activity, and monocyte/macrophage progenitor cell proliferation cytokines in head kidney cells under immune stimulatory conditions. The expression profiles were dissimilar in the unstimulated control and immune-stimulated cells. Moreover, increased expression profile was observed due to different stimulations for IL-1b, IL-6, IL-10, IL-12p35, IL-12p40, IL-21, TNF-a, TNF-N, I-IFN-1 and IFN-g genes. These results suggest that cytokine genes could be used as biomarkers to know the innate immune status of fish.