Chitosan Treatment of E-11 Cells Modulates Transcription of Nonspecific Immune Genes and Reduces Nodavirus Capsid Protein Gene Expression

This study explores whether crustacean products inhibit viral infections in aquaculture. Chitosan (CHT) was extracted from waste products of <i>Parapenaeus longirostris</i>. Biochemical composition, viscosity measurement, molecular weight, structure and cytotoxicity tests were used to ch...

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Autores principales:	Nadia Chérif, Fatma Amdouni, Boutheina Bessadok, Ghada Tagorti, Saloua Sadok
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	nodavirus chitosan gene expression prevention Veterinary medicine SF600-1100 Zoology QL1-991
Acceso en línea:	https://doaj.org/article/37b03b48577b47d6986dca72a103d8ae
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spelling	oai:doaj.org-article:37b03b48577b47d6986dca72a103d8ae2021-11-25T16:15:13ZChitosan Treatment of E-11 Cells Modulates Transcription of Nonspecific Immune Genes and Reduces Nodavirus Capsid Protein Gene Expression10.3390/ani111130972076-2615https://doaj.org/article/37b03b48577b47d6986dca72a103d8ae2021-10-01T00:00:00Zhttps://www.mdpi.com/2076-2615/11/11/3097https://doaj.org/toc/2076-2615This study explores whether crustacean products inhibit viral infections in aquaculture. Chitosan (CHT) was extracted from waste products of <i>Parapenaeus longirostris</i>. Biochemical composition, viscosity measurement, molecular weight, structure and cytotoxicity tests were used to characterize the extracted chitosan. Cultures of E-11 cells derived from snakehead <i>Ophicephalus striatus</i> were inoculated with 10<sup>6.74</sup> TCID<sub>50</sub> of an isolate of betanodavirus genotype RGNNV (redspotted grouper nervous necrosis virus) after being treated with solutions of 0.3% CHT for 1 h at room temperature. The antiviral effect of CHT was assessed by comparing the ability of RGNVV to replicate and produce cytopathic effects on CHT-treated cell cultures. The change in RNA expression levels of the nodavirus capsid protein gene and three mediator genes in infected cells with or without CHT treatment was evaluated by qPCR. Changes in gene expression compared to control groups were monitored at 6, 24, 48 and 71 h post treatment in all target gene transcripts. The CCR3 expression in CHT treated cells showed a significant increase (<i>p</i> < 0.05) until day 3. On the other hand, the expression of TNF-α decreased significantly (<i>p</i> < 0.05) in CHT treated cells throughout the experimental period. Likewise, the expression of the IL-10 gene showed a significant downregulation in CHT treated cells at all time points (<i>p</i> ≤ 0.05). As further evidence of an antiviral effect, CHT treatment of cells produced a reduction in virus load as measured by a reduced expression of the viral capsid gene and the increase in RQ values from 406 ± 1.9 at hour 1 to 695 ± 3.27 at 72 h post inoculation. Statistical analysis showed that the expression of the viral capsid gene was significantly lower in cells treated with chitosan (<i>p</i> ≤ 0.05). These results improve our knowledge about the antiviral activity of this bioactive molecule and highlight its potential use in fish feed industry.Nadia ChérifFatma AmdouniBoutheina BessadokGhada TagortiSaloua SadokMDPI AGarticlenodaviruschitosangene expressionpreventionVeterinary medicineSF600-1100ZoologyQL1-991ENAnimals, Vol 11, Iss 3097, p 3097 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	nodavirus chitosan gene expression prevention Veterinary medicine SF600-1100 Zoology QL1-991
spellingShingle	nodavirus chitosan gene expression prevention Veterinary medicine SF600-1100 Zoology QL1-991 Nadia Chérif Fatma Amdouni Boutheina Bessadok Ghada Tagorti Saloua Sadok Chitosan Treatment of E-11 Cells Modulates Transcription of Nonspecific Immune Genes and Reduces Nodavirus Capsid Protein Gene Expression
description	This study explores whether crustacean products inhibit viral infections in aquaculture. Chitosan (CHT) was extracted from waste products of <i>Parapenaeus longirostris</i>. Biochemical composition, viscosity measurement, molecular weight, structure and cytotoxicity tests were used to characterize the extracted chitosan. Cultures of E-11 cells derived from snakehead <i>Ophicephalus striatus</i> were inoculated with 10<sup>6.74</sup> TCID<sub>50</sub> of an isolate of betanodavirus genotype RGNNV (redspotted grouper nervous necrosis virus) after being treated with solutions of 0.3% CHT for 1 h at room temperature. The antiviral effect of CHT was assessed by comparing the ability of RGNVV to replicate and produce cytopathic effects on CHT-treated cell cultures. The change in RNA expression levels of the nodavirus capsid protein gene and three mediator genes in infected cells with or without CHT treatment was evaluated by qPCR. Changes in gene expression compared to control groups were monitored at 6, 24, 48 and 71 h post treatment in all target gene transcripts. The CCR3 expression in CHT treated cells showed a significant increase (<i>p</i> < 0.05) until day 3. On the other hand, the expression of TNF-α decreased significantly (<i>p</i> < 0.05) in CHT treated cells throughout the experimental period. Likewise, the expression of the IL-10 gene showed a significant downregulation in CHT treated cells at all time points (<i>p</i> ≤ 0.05). As further evidence of an antiviral effect, CHT treatment of cells produced a reduction in virus load as measured by a reduced expression of the viral capsid gene and the increase in RQ values from 406 ± 1.9 at hour 1 to 695 ± 3.27 at 72 h post inoculation. Statistical analysis showed that the expression of the viral capsid gene was significantly lower in cells treated with chitosan (<i>p</i> ≤ 0.05). These results improve our knowledge about the antiviral activity of this bioactive molecule and highlight its potential use in fish feed industry.
format	article
author	Nadia Chérif Fatma Amdouni Boutheina Bessadok Ghada Tagorti Saloua Sadok
author_facet	Nadia Chérif Fatma Amdouni Boutheina Bessadok Ghada Tagorti Saloua Sadok
author_sort	Nadia Chérif
title	Chitosan Treatment of E-11 Cells Modulates Transcription of Nonspecific Immune Genes and Reduces Nodavirus Capsid Protein Gene Expression
title_short	Chitosan Treatment of E-11 Cells Modulates Transcription of Nonspecific Immune Genes and Reduces Nodavirus Capsid Protein Gene Expression
title_full	Chitosan Treatment of E-11 Cells Modulates Transcription of Nonspecific Immune Genes and Reduces Nodavirus Capsid Protein Gene Expression
title_fullStr	Chitosan Treatment of E-11 Cells Modulates Transcription of Nonspecific Immune Genes and Reduces Nodavirus Capsid Protein Gene Expression
title_full_unstemmed	Chitosan Treatment of E-11 Cells Modulates Transcription of Nonspecific Immune Genes and Reduces Nodavirus Capsid Protein Gene Expression
title_sort	chitosan treatment of e-11 cells modulates transcription of nonspecific immune genes and reduces nodavirus capsid protein gene expression
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/37b03b48577b47d6986dca72a103d8ae
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Chitosan Treatment of E-11 Cells Modulates Transcription of Nonspecific Immune Genes and Reduces Nodavirus Capsid Protein Gene Expression

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