日本牙鲆 Toll 样受体 3 的分子克隆与特性分析
Molecular cloning and characterization of Toll-like receptor 3 in Japanese flounder, Paralichthys olivaceus.
机构信息
Laboratory of Genome Science, Tokyo University of Marine Science and Technology, Konan 4-5-7, Minato-ku, Tokyo 108-8477, Japan.
出版信息
Dev Comp Immunol. 2012 May;37(1):87-96. doi: 10.1016/j.dci.2011.12.004. Epub 2011 Dec 19.
Mammalian Toll-like receptor 3 (TLR3) recognizes extracellular and intracellular viral dsRNA, and then initiates signaling cascades leading to NF-κB activation and interferon (IFN) production. To understand the roles of TLR3 in the fish immune system, TLR3 gene (JfTLR3) was identified from Japanese flounder (Paralichthys olivaceus), which consisted of 4 exons and 3 introns. Its expression in peripheral blood leukocytes increased upon stimulation with poly I:C and CpG ODN 1668. Exposure to viral hemorrhagic septicemia virus increased expression of JfTLR3 in the blood, liver, head kidney and spleen. Intracellular poly I:C stimulation in JfTLR3-overexpressing YO-K cells significantly induced IFN-inducible and NF-κB-regulated genes. NF-κB activity in JfTLR3-overexpressing YO-K cells was significantly induced by intracellular poly I:C while expression of IFN-inducible genes and NF-κB reporter activity in JfTLR3-overexpressing HINAE cells increased upon stimulation by extracellular poly I:C. These results suggest that JfTLR3 plays an important role in the induction of antiviral immune response.
哺乳动物 Toll 样受体 3（TLR3）识别细胞外和细胞内的病毒 dsRNA，然后启动信号级联反应，导致 NF-κB 激活和干扰素（IFN）的产生。为了了解 TLR3 在鱼类免疫系统中的作用，从日本牙鲆（Paralichthys olivaceus）中鉴定出 TLR3 基因（JfTLR3），它由 4 个外显子和 3 个内含子组成。其在外周血白细胞中的表达在受到 poly I:C 和 CpG ODN 1668 的刺激后增加。病毒出血性败血症病毒的暴露导致血液、肝脏、头肾和脾脏中 JfTLR3 的表达增加。在过表达 JfTLR3 的 YO-K 细胞中，细胞内 poly I:C 的刺激显著诱导 IFN 诱导和 NF-κB 调节基因的表达。细胞内 poly I:C 显著诱导过表达 JfTLR3 的 YO-K 细胞中的 NF-κB 活性，而在过表达 JfTLR3 的 HINAE 细胞中，IFN 诱导基因的表达和 NF-κB 报告基因活性在受到细胞外 poly I:C 的刺激后增加。这些结果表明 JfTLR3 在诱导抗病毒免疫反应中起着重要作用。
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