暗鲀12种白细胞介素的克隆、鉴定及基因表达分析

Ying Huang , Peng Luo , Fu-Hui Jiang , Hui-Ze Gao , Li-Fan Cui , Zhe Zhao
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摘要

白细胞介素(IL)是分泌型细胞因子的一个亚群,是参与免疫系统细胞间调节的分子。在本研究中,从模糊河豚中克隆并功能鉴定了12个IL同源物,它们被称为ToIL-1β、ToIL-1、ToIL-6、ToIL-10、ToIL-11、ToIL-12、ToIL-17、ToIL-18、ToIL-20、ToIL-24、ToIL-27和ToIL-34。多重比对结果表明,除了ToIL-24和ToIL-27外,其他推导的ToIL蛋白与其他已知的鱼类IL具有典型的特征和结构。系统发育分析表明,12个ToIL在进化上与其他脊椎动物的同类关系密切。组织分布测定表明,大多数ToIL基因的mRNA转录物在所有检查的组织中都是组成性表达的,在免疫组织中表达相对较高。哈维氏弧菌和金黄色葡萄球菌感染后,脾脏和肝脏中12种ToIL的表达水平显著上调,并且它们的反应随时间变化。总之,这些数据与ToIL的表达和在不同测试情况下的免疫反应进行了相应的讨论。结果表明,这12个ToIL基因参与了蒙氏锥虫的抗菌免疫反应。
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Molecular cloning, characterization and gene expression analysis of twelve interleukins in obscure puffer Takifugu obscurus

Interleukins (ILs) are a subgroup of secreted cytokines, which are molecules involved in the intercellular regulation of the immune system. In this study, 12 IL homologs were cloned and functionally identified from obscure puffer Takifugu obscurus, and they were termed as ToIL-1β, ToIL-1, ToIL-6, ToIL-10, ToIL-11, ToIL-12, ToIL-17, ToIL-18, ToIL-20, ToIL-24, ToIL-27, and ToIL-34. Multiple alignment results showed that except for ToIL-24 and ToIL-27, other deduced ToIL proteins shared typical characteristics and structure with other known fish ILs. Phylogenetic analysis revealed that 12 ToILs were evolutionarily closely related to their counterparts in other selected vertebrates. Tissue distribution assay demonstrated that the mRNA transcripts of most ToIL genes were constitutively expressed in all tissues examined, with relatively high expression in immune tissues. Following Vibrio harveyi and Staphylococcus aureus infection, the expression levels of 12 ToILs in the spleen and liver were significantly upregulated, and their response over time varied. Taken together, these data were discussed accordingly with the ToIL expression and the immune response under the different situations tested. The results suggest that the 12 ToIL genes are involved in the antibacterial immune response in T. obscurus.

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