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具有抗病毒活性的抗菌肽(AMP)对鱼类诺达病毒。

Antimicrobial peptides (AMP) with antiviral activity against fish nodavirus.

机构信息

Institute of Zoology, National Taiwan University, Taipei 106, Taiwan.

出版信息

Fish Shellfish Immunol. 2010 Mar;28(3):434-9. doi: 10.1016/j.fsi.2009.11.020. Epub 2009 Dec 8.

Abstract

Nervous necrosis virus (NNV) is classified as betanodavirus of Nodaviridae, and has caused mass mortality of numerous marine fish species at larval stage. Antimicrobial peptides (AMPs) play an important role of innate immunity either against bacterial pathogens or viruses. Up to date, little is known if any AMP could effectively inhibit fish nodaviruses and its mechanism. In this study, the antiviral activities of three antimicrobial peptides (AMPs) against grouper NNV (GNNV) were screened in the fish cell line. Two of the three AMPs, tilapia hepcidin 1-5 (TH 1-5) and cyclic shrimp anti-lipopolysaccharide factor (cSALF), were able to agglutinate purified NNV particles into clump, and the clumps were further confirmed to be viral proteins by TEM and Western blot. The NNV solution, separately pre-mixed with AMP (TH 1-5 or cSALF) or deionized-distilled water for 1 h, was used to infect GF-1 cells, and the levels of capsid protein in the GNNV-AMP-infected cells at 1 h post infection were much lower than that in the GNNV-H(2)O-infected cells, indicating that only a small portion of viral particles in the GNNV-AMP mixture could successfully infected the cells. Treatment of cBB cells with TH 1-5 and cSALF did not induce Mx gene expression; however, grouper epinecidin-1 (CP643-1) could induce the expression of Mx in the pre-treated cBB cells. This study revealed three AMPs with anti-NNV activity through two different mechanisms, and shed light on the future application in aquaculture.

摘要

神经坏死病毒(NNV)被归类为诺达病毒科的贝塔诺病毒,已导致许多海洋鱼类在幼虫阶段大量死亡。抗菌肽(AMPs)在对抗细菌病原体或病毒的固有免疫中发挥着重要作用。迄今为止,尚不清楚是否有任何 AMP 可以有效抑制鱼类诺达病毒及其机制。在这项研究中,在鱼类细胞系中筛选了三种抗菌肽(AMPs)对石斑鱼神经坏死病毒(GNNV)的抗病毒活性。这三种 AMP 中的两种,罗非鱼肝抗菌肽 1-5(TH 1-5)和环状虾抗脂多糖因子(cSALF),能够将纯化的 NNV 颗粒凝集成团,并且通过 TEM 和 Western blot 进一步证实这些团块是病毒蛋白。将 AMP(TH 1-5 或 cSALF)或去离子蒸馏水分别与 NNV 溶液预混合 1 h,然后用于感染 GF-1 细胞,感染后 1 h,在 GNNV-AMP 感染细胞中的衣壳蛋白水平明显低于 GNNV-H2O 感染细胞中的衣壳蛋白水平,这表明在 GNNV-AMP 混合物中只有一小部分病毒颗粒能够成功感染细胞。用 TH 1-5 和 cSALF 处理 cBB 细胞不会诱导 Mx 基因表达;然而,石斑鱼 epinecidin-1(CP643-1)可以诱导预处理的 cBB 细胞中 Mx 的表达。这项研究揭示了三种具有抗 NNV 活性的 AMP 通过两种不同的机制,为水产养殖的未来应用提供了思路。

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