Molecular and functional analysis of an interferon gene from the zebrafish, Danio rerio
Molecular and functional analysis of an interferon gene from the zebrafish, Danio rerio
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DOI：
10.1128/jvi.77.3.1992-2002.2003
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发表时间：
2003-02-01
影响因子：
5.4
通讯作者：
Kim, CH
中科院分区：
文献类型：
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作者：
Altmann, SM;Mellon, MT;Kim, CH
由α IFN（IFN-α）、IFN-β、IFN-ω、IFN-δ、IFN-κ和IFN-τ组成的干扰素（IFN）家族是参与针对各种微生物的先天免疫应答的一大组细胞因子。IFN基因已经从多种哺乳动物和鸟类中克隆出来，然而，来自低等脊椎动物的IFN基因还没有出现。在这里，我们报告的干扰素基因的克隆和表征从斑马鱼，斑马鱼。斑马鱼IFN（zfIFN）的长度为185个氨基酸，其中前22个氨基酸代表推定的信号肽。用已知的IFN诱导剂聚肌苷酸-聚胞苷酸（poly[I]-poly[C]）处理导致zfIFN mRNA转录物的增加。zfIFN也能够激活IFN诱导型Mx启动子时，共转染的质粒含有斑马鱼Mx启动子上游的荧光素酶报告基因。为了证明抗病毒活性，用zfIFN转染斑马鱼细胞并用鱼弹状病毒攻击。与用对照载体转染的细胞相比，在znIFN转染的细胞中观察到噬斑数目减少36%。系统发育分析表明zfIFN与禽类和哺乳动物IFN的分化程度大致相同，这与其起源于这些分化谱系的最近共同祖先中存在的IFN一致。当使用zfIFN搜索河豚基因组数据库时，还发现了来自河豚的推定IFN，红鳍东方鲀，这表明zfIFN可用于寻找其他鱼类IFN基因。这些结果表明，斑马鱼可以作为研究先天免疫和感染性疾病免疫反应的有效模型。
The interferon (IFN) family consisting of alpha IFN (IFN-alpha), IFN-beta, IFN-omega, IFN-delta, IFN-kappa, and IFN-tau is a large group of cytokines involved in the innate immune response against various microorganisms. Genes for IFN have been cloned from a variety of mammalian and avian species; however, IFN genes from lower-order vertebrates have not been forthcoming. Here, we report the cloning and characterization of the IFN gene from the zebrafish, Danio rerio. Zebrafish IFN (zfIFN) is 185 amino acids in length, with the first 22 amino acids representing a putative signal peptide. Treatment with the known IFN inducer polyinosinic acid-polycytidylic acid (poly[I]-poly[C]) resulted in an increase in zfIFN mRNA transcripts. zfIFN was also able to activate the IFN-inducible Mx promoter when cotransfected with a plasmid containing the zebrafish Mx promoter upstream of a luciferase reporter gene. To demonstrate antiviral activity, zebrafish cells were transfected with zfIFN and challenged with a fish rhabdovirus. A 36% reduction in plaque number was seen in znIFN-transfected cells, compared to cells transfected with a control vector. Phylogenetic analysis has shown zfIFN to be approximately equally divergent from avian and mammalian IFN, consistent with its origin from an IFN present in the most recent common ancestor of these divergent lineages. A putative IFN from puffer, Fugu rubripes, was also found when zfIFN was used to search the fugu genome database, demonstrating that zfIFN can be used to find additional fish IFN genes. These results demonstrate that zebrafish can be used as an effective model for studying innate immunity and immune response to infectious disease.