Identitication and expression analysis of lymphotoxin-beta like hornologues in rainbow trout Oncorhynchus mykiss
Identitication and expression analysis of lymphotoxin-beta like hornologues in rainbow trout Oncorhynchus mykiss
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DOI：
10.1016/j.molimm.2005.07.037
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发表时间：
2006-03-01
影响因子：
3.6
通讯作者：
Secombes, CJ
中科院分区：
文献类型：
--
作者：
Kono, T;Zou, J;Secombes, CJ
一种淋巴毒素- β (lt - β)基因在虹鳟鱼中被克隆并测序，为硬骨鱼中存在lt - β提供了第一个确凿的证据。分离出两种lt - β亚型。lt - β 1 cDNA由952 bp（含139 bp的5'-UTR和201 bp的3'-UTR）和lt - β 2 cDNA由836 bp（含237 bp的5'-UTR和197 bp的3'-UTR）组成，两者均可翻译成含有203个氨基酸残基的蛋白。这两种异构体都含有预测的21个氨基酸残基的跨膜结构域(Leu（11)-Val(31)）和TNF家族特征(Val（104)-Phe(120)）。鳟鱼与其他已知TNF家族成员的同源性和系统发育分析显示，鳟鱼与TNF- n（硬骨鱼）和其他TNF- β（哺乳动物和青蛙）具有良好的相似性。lt - β 1和tnf - α（1和2）基因在未受刺激的鳟鱼头肾、脾脏、鳃和肠中高表达，而lt - β 2仅在鳃中弱表达。巨噬细胞（RTS-11）和成纤维细胞（RTG-2）样细胞系中均未发现lt - β 1和- β 2基因的表达，但tnf - α 2基因在两种细胞系中均有表达，tnf - α 1基因仅在RTS-11细胞中表达。在头肾细胞中，PHA或LPS刺激可增加lt - β 1和tnf - α(1,2)基因的表达。鳟鱼lt - β的发现将允许对鱼类炎症反应进行更完整的分析。(c) 2005 Elsevier Ltd版权所有。
A lymphotoxin-beta (LT-beta) gene has been cloned and sequenced in rainbow trout and provides the first conclusive evidence for the existence of LT-beta in teleost. Two isoforms of LT-beta were isolated. LT-beta 1 cDNA was composed of 952 bp (with a 139 bp 5'-UTR and a 201 bp 3'-UTR) and LT-beta 2 cDNA was 836 bp (with a 237 bp 5'-UTR and a 197 bp 3'-UTR) both of which translated into a protein of 203 amino acid residues. Both isoforms contained a predicted transmembrane domain of 21 amino acid residues (Leu(11)-Val(31)) and the TNF family signature (Val(104)-Phe(120)). Homology and phylogenetic analysis of trout LT-beta's with other known TNF family member showed good similarity to TNF-N (teleost) and other LT-beta (mammals and frog). LT-beta 1 and TNF-alpha (1 and 2) genes were highly expressed in unstimulated trout head kidney, spleen, gill and intestine, whereas LT-beta 2 was weakly expressed only in the gill. The expression of LT-beta 1 and -beta 2 genes was not found in macrophage (RTS-11) and fibroblast (RTG-2) like cell lines, although the TNF-alpha 2 gene was expressed in both cell lines with the TNF-alpha 1 gene only expressed in RTS-11 cells. In head kidney cells, expression of LT-beta 1 and TNF-alpha (1, 2) genes was increased by stimulation with PHA or LPS. The discovery of trout LT-beta will allow a more complete analysis of fish inflammatory responses. (c) 2005 Elsevier Ltd. All rights reserved.