Kono Tomoya, Takayama Hiroaki, Nagamine Ryusuke, Korenaga Hiroki, Sakai Masahiro
Interdisciplinary Research Organization, University of Miyazaki, 1-1 Gakuen kibanadai-nishi, Miyazaki 889-2192, Japan.
Vet Immunol Immunopathol. 2013 Jan 15;151(1-2):90-101. doi: 10.1016/j.vetimm.2012.10.012. Epub 2012 Nov 16.
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 (IL-1β, IL-6, IL-17A/F3, IL-18, TNF-α, TNF-N), anti-inflammatory (IL-4/13A, IL-4/13B, IL-10), T-cell proliferation/differentiation (IL-2, IL-15, IL-21, TGF-β1), B-cell activation/differentiation (IL-7, IL-6, IL-4/13A, IL-4/13B), NK cell stimulation (IL-12p35 and IL-12p40), induction of anti-viral activity (I-IFN-1 and IFN-γ), and monocyte/macrophage progenitor cell proliferation (M-CSF1b) 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-1β, IL-6, IL-10, IL-12p35, IL-12p40, IL-21, TNF-α, TNF-N, I-IFN-1 and IFN-γ genes. These results suggest that cytokine genes could be used as biomarkers to know the immune status of fish. The constructed multiplex RT-PCR assay will enhance understanding on immune regulation by cytokines in fish.
为监测日本河豚细胞因子基因的表达，开发了一种用于定量多重分析的新型平台。这种定制设计的多重逆转录聚合酶链反应（RT-PCR）检测方法用于分析19种细胞因子基因的表达谱，包括促炎细胞因子（白细胞介素-1β、白细胞介素-6、白细胞介素-17A/F3、白细胞介素-18、肿瘤坏死因子-α、肿瘤坏死因子-N）、抗炎细胞因子（白细胞介素-4/13A、白细胞介素-4/13B、白细胞介素-10）、T细胞增殖/分化相关细胞因子（白细胞介素-2、白细胞介素-15、白细胞介素-21、转化生长因子-β1）、B细胞活化/分化相关细胞因子（白细胞介素-7、白细胞介素-6、白细胞介素-4/13A、白细胞介素-4/13B）、自然杀伤（NK）细胞刺激相关细胞因子（白细胞介素-12p35和白细胞介素-12p40）、抗病毒活性诱导相关细胞因子（I型干扰素-1和干扰素-γ）以及单核细胞/巨噬细胞祖细胞增殖相关细胞因子（巨噬细胞集落刺激因子1b）在免疫刺激条件下头部肾细胞中的表达情况。未刺激的对照细胞和免疫刺激细胞中的表达谱不同。此外，由于对白细胞介素-1β、白细胞介素-6、白细胞介素-10、白细胞介素-12p35、白细胞介素-12p40、白细胞介素-21、肿瘤坏死因子-α、肿瘤坏死因子-N、I型干扰素-1和干扰素-γ基因的不同刺激，观察到表达谱增加。这些结果表明，细胞因子基因可作为了解鱼类免疫状态的生物标志物。构建的多重RT-PCR检测方法将增进对鱼类细胞因子免疫调节的理解。
