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Ctn[15-34](一种源自组织蛋白酶原的二十碳肽)对凡纳滨对虾原代血细胞培养物中传染性肌肉坏死病毒的抗病毒活性

Antiviral Activity of Ctn[15-34], A Cathelicidin-Derived Eicosapeptide, Against Infectious Myonecrosis Virus in Litopenaeus vannamei Primary Hemocyte Cultures.

作者信息

Vieira-Girão P R N, Falcão C B, Rocha I R C B, Lucena H M R, Costa F H F, Rádis-Baptista G

机构信息

Laboratory of Biochemistry and Biotechnology, Institute for Marine Sciences, Federal University of Ceará, Av. da Abolição 3207, Fortaleza, CE, 60165-081, Brazil.

Northeast Biotechnology Network (RENORBIO), Post-graduation program in Biotechnology, Federal University of Ceará, Fortaleza, CE, 60455-900, Brazil.

出版信息

Food Environ Virol. 2017 Sep;9(3):277-286. doi: 10.1007/s12560-017-9285-5. Epub 2017 Feb 16.

Abstract

The shrimp farming has been converted into a mature aquaculture industry dealing with over millions of metric tonnes of processed commodities. Nevertheless, the global shrimp productions are constantly threatened by disease outbreaks, mainly triggered by rapidly disseminating viruses. Infectious myonecrosis virus (IMNV) is one of these epizootic agents affecting shrimp production in Brazil, of which no treatment exists. Herein, the antiviral activity against IMNV of an eicosapeptide, named Ctn[15-34], derived from a member of the cathelicidin family of antimicrobial peptides, was demonstrated. Cultures of hemocytes from Litopenaeus vannamei were established that support IMNV replication and infectivity titration. The cytotoxic effect of IMNV in culture and the in vitro anti-IMNV activity of Ctn[15-34] were assessed using a high-sensitive fluorescent-based method in combination with quantitative PCR. The Ctn[15-34] (<12.5 µM) neutralized the toxic effects of IMNV at loads sufficient to kill 50% of shrimp hemocytes. This study reported for the first time the replication of IMNV in vitro and the employment of a straightforward methodology to assess cell viability and viral/antiviral activities. In addition, it provided the basis for the development of the anti-infective multi-effector Ctn[15-34] eicosapeptide and analogs as components of antiviral formulations against shrimp viral diseases.

摘要

对虾养殖已发展成为一个成熟的水产养殖业,涉及加工量超过数百万吨的商品。然而,全球对虾产量一直受到疾病爆发的威胁,主要是由迅速传播的病毒引发的。传染性肌肉坏死病毒(IMNV)是影响巴西对虾生产的这些流行性病原体之一,目前尚无治疗方法。在此,证明了一种源自抗菌肽cathelicidin家族成员的二十肽Ctn[15-34]对IMNV具有抗病毒活性。建立了凡纳滨对虾血细胞培养物,用于支持IMNV复制和感染性滴定。使用基于高灵敏度荧光的方法结合定量PCR评估了IMNV在培养物中的细胞毒性作用以及Ctn[15-34]的体外抗IMNV活性。Ctn[15-34](<12.5 μM)在足以杀死50%对虾血细胞的病毒载量下中和了IMNV的毒性作用。本研究首次报道了IMNV在体外的复制以及采用一种简单方法来评估细胞活力和病毒/抗病毒活性。此外,它为开发抗感染多效应二十肽Ctn[15-34]及其类似物作为针对对虾病毒疾病的抗病毒制剂成分提供了依据。

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