Seppola Marit, Larsen Atle Noralf, Steiro Kari, Robertsen Børre, Jensen Ingvill
Fiskeriforskning, Norwegian Institute of Fisheries and Aquaculture Research, N-9291 Tromsø, Norway.
Mol Immunol. 2008 Feb;45(4):887-97. doi: 10.1016/j.molimm.2007.08.003. Epub 2007 Sep 17.
The mammalian interleukins IL-1beta and IL-8 are important pro-inflammatory cytokines often used as markers of an activated inflammatory response, while IL-10 is regarded as an anti-inflammatory cytokine and plays a crucial role in the regulation of inflammation. Few cytokines from gadoid fish have been described, and herein the sequence and expression of these interleukin genes are studied in Atlantic cod (Gadus morhua L.). IL-1beta, IL-8 and IL-10 from cod show similarities in gene organisation and predicted protein sequences, compared to counterpart genes in other teleosts. Gene expression was studied using quantitative real time PCR in response to several treatments both in vitro and in vivo. In adherent head kidney cells, infectious pancreatic necrosis virus (IPNV) and lipopolysaccharide (LPS) significantly stimulated gene expression of IL-1beta. The expression of IL-1beta was not increased after treatment with a viral imitator (poly I:C), and neither IL-8 nor IL-10 responded to any of these agents in vitro. However, in vivo administrated poly I:C and formalin-killed Vibrio anguillarum (In-V.ang) induced interleukin expression, varying in intensity between different organs. IL-1beta and IL-10 gene expression profiles showed an opposite induction pattern in the in vivo experiments. Injection of In-V.ang highly induced IL-1beta expression, while a low induction was evident for IL-10, whereas the opposite was observed after injection of poly I:C. This pattern was particularly marked in spleen, where also IL-8 followed the expression pattern of IL-1beta. The opposite expression profiles indicate a suppressive role for IL-10 on the transcription of IL-1beta, and to a lesser extent on IL-8 transcription. Along with the identification of important promoter regulatory motives, these results provide tools for studying inflammatory responses in cod.
哺乳动物白细胞介素IL - 1β和IL - 8是重要的促炎细胞因子,常被用作激活炎症反应的标志物,而IL - 10被视为抗炎细胞因子,在炎症调节中起关键作用。关于鳕形目鱼类细胞因子的描述较少,本文研究了大西洋鳕鱼(Gadus morhua L.)中这些白细胞介素基因的序列和表达。与其他硬骨鱼的对应基因相比,鳕鱼的IL - 1β、IL - 8和IL - 10在基因结构和预测的蛋白质序列上具有相似性。使用定量实时PCR研究了体外和体内几种处理后的基因表达。在贴壁的头肾细胞中,传染性胰腺坏死病毒(IPNV)和脂多糖(LPS)显著刺激了IL - 1β的基因表达。用病毒模拟物(聚肌胞苷酸,poly I:C)处理后,IL - 1β的表达没有增加,体外实验中IL - 8和IL - 10对任何这些试剂均无反应。然而,体内给予poly I:C和福尔马林灭活的鳗弧菌(In - V.ang)可诱导白细胞介素表达,不同器官的诱导强度不同。在体内实验中,IL - 1β和IL - 10基因表达谱呈现相反的诱导模式。注射In - V.ang可高度诱导IL - 1β表达,而对IL - 10的诱导作用较弱,而注射poly I:C后则观察到相反的情况。这种模式在脾脏中尤为明显,其中IL - 8也遵循IL - 1β的表达模式。相反的表达谱表明IL - 10对IL - 1β的转录具有抑制作用,对IL - 8转录的抑制作用较小。随着重要启动子调控基序的鉴定,这些结果为研究鳕鱼的炎症反应提供了工具。