Studies on the structure and function of tumor necrosis factor (TNF) in fish

鱼类肿瘤坏死因子（TNF）的结构与功能研究

• 批准号: 15380136
• 负责人: NAKANISHI Teruyuki
• 金额: $ 9.47万
• 依托单位: Nihon University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15380136/
• 关键词: Rainbow trout; Zebrafish; Ayu fish; Banded dogfish; TNF alpha; Transgenic; mRNA expression; Recombinant TNF; GFP; 組換えTNF; TNFレセプターデコイ; LPS; 定量PCR法; 組換えニジマスTNF; CCケモカイン

Tumor necrosis factor (TNF) is a primary mediator of immune regulation and the inflammatory response. TNF is also known to be a mediator involved in the interaction between endocrine nervous and immune systems. There have been also reported that TNF/LT family cytokines play important role in development, maturation and maintenance of secondary lymphoid organs. Thus TNF is one of the most important cytokines investigated so far from various fields and aspects.To understand the broad range of biological activities of TNF we tried to investigate its pleiotropic nature from phylogenetical and ontogenetical points of view. In the present research we obtained the following results :1.Structure and function of fish TNFWe isolated one TNF homologue from Ayu fish and two types of TNF from rainbow trout and zebrafish. We also cloned TNF decoy receptor 3 (DcR3) together with several cytokines from cartilaginous fish. Structure and expression of these fish TNF and TNF super family genes resembles to those of mammalian TNF.2.Production of recombinant TNF and its biological activityPurified recombinant TNF of Ayu fish induced the production of superoxide from head-kidney leukocytes.3.Expression of TNF mRNA after stimulation with LPS and/or bacteriaExpression of TNF in tissues after stimulation with LPS was different between two types of TNF in rainbow trout. Studies using transgenic rainbow trout introduced with TNF promoter linked to green fluorescence protein (GFP) as a reporter gene showed that TNF secreting cells are involved in envelopment response against bacteria.4.Role of TNF in developmentTwo types of TNF were weakly expressed at the nervous system of zebrafish embryos, and increased TNF expression toward hatching was observed in Ayu embryos suggesting the role of TNF in the morphogenesis of embryonic organs.

肿瘤坏死因子（TNF）是免疫调节和炎症反应的主要介质。 TNF 还被认为是参与内分泌神经和免疫系统之间相互作用的介质。也有报道称TNF/LT家族细胞因子在次级淋巴器官的发育、成熟和维持中发挥着重要作用。因此，TNF是迄今为止从各个领域和方面研究的最重要的细胞因子之一。为了了解TNF的广泛生物活性，我们试图从系统发育和个体发育的角度研究其多效性。在本研究中我们得到了以下结果： 1.鱼类TNF的结构和功能我们从香鱼中分离到一种TNF同源物，从虹鳟鱼和斑马鱼中分离到两种TNF同源物。我们还克隆了 TNF 诱饵受体 3 (DcR3) 以及来自软骨鱼的几种细胞因子。这些鱼类TNF和TNF超家族基因的结构和表达与哺乳动物TNF相似。2.重组TNF的产生及其生物活性纯化的香鱼重组TNF诱导头肾白细胞产生超氧化物。3.TNF的表达用LPS和/或细菌刺激后的mRNA用LPS刺激后组织中TNF的表达在虹鳟鱼中两种类型的TNF之间是不同的。使用导入连接有绿色荧光蛋白（GFP）作为报告基因的TNF启动子的转基因虹鳟鱼的研究表明，TNF分泌细胞参与针对细菌的包膜反应。 4.TNF在发育中的作用两种类型的TNF在神经细胞中弱表达在斑马鱼胚胎系统中，在香鱼胚胎中观察到孵化过程中 TNF 表达增加，表明 TNF 在胚胎器官形态发生中的作用。

项目成果

Yoshimura, H., Dhar, DK., Nakamoto, T., Kotoh, T., Takano, M., Soma, G-I, Nagasue, N: "Prognostic significance of tumor necrosis factor receptor in colorectal adenocarcinoma"Anticancer Research. 23. 85-89 (2003)

Yoshimura, H.、Dhar, DK.、Nakamoto, T.、Kotoh, T.、Takano, M.、Soma, G-I、Nagasue, N：“肿瘤坏死因子受体在结直肠腺癌中的预后意义”抗癌研究。

Molecular cloning and preliminary expression analysis of banded dogfish (Triakis scyllia) CC chemokine cDNAs by use of suppression subtractive hybridization.

利用抑制消减杂交对带状角鲨 (Triakis scyllia) CC 趋化因子 cDNA 进行分子克隆和初步表达分析。

• 发表时间: 2005
• 期刊: Immunogenetics 56
• 作者: T.Gao;Y.Wng;J.Liu;S.Watanabe;R.Fuseya;伏屋 玲子;Natsuko Kajiwara et al.;Natsuko Kajiwara et al.;Karri Ramu et al.;Inoue Yuuki
• 通讯作者: Inoue Yuuki

Inherent potential for production of tumor necrosis factor-□ by human intestinal macrophages.

人类肠道巨噬细胞产生肿瘤坏死因子-□的固有潜力。

• 发表时间: 2006
• 期刊: Int J Colorectal Dis. 21
• 作者: Nakata;K.;H.Inagawa;T.Nishizawa;T.Honda;C.Kohchi;Y.Tonomoto;H.Yoshimura;N.Nagase;S.Natori;H.Terada;G-I.Soma
• 通讯作者: G-I.Soma

p, p'-DDE depresses the immune competence of chinook salmon (Oncorhynchus tshawytscha) leucocytes.

p, p-DDE 会抑制大鳞大麻哈鱼 (Oncorhynchus tshawytscha) 白细胞的免疫能力。

• 发表时间: 2005
• 期刊: Fish & Shellfish Immunol. 19(2)
• 作者: Misumi I.;A.T.Vella;J.C.Leongi;T.Nakanishi;C.B.Schreck
• 通讯作者: C.B.Schreck

Yoshiura, Y., I.Kiryu, A.Fujiwara, H.Suetake, Y.Suzuki, T.Nakanishi, M.Ototake: "Identification and characterization of Fugu orthologues of mammalian interleukin-12 subunits"Immunogenetics. 55. 296-306 (2003)

Yoshiura, Y., I.Kiryu, A.Fujiwara, H.Suetake, Y.Suzuki, T.Nakanishi, M.Ototake：“哺乳动物白细胞介素 12 亚基的河豚直系同源物的鉴定和表征”免疫遗传学。