Effect of double-stranded RNA and interferon on the antiviral activity of Atlantic salmon cells against infectious salmon anemia virus and infectious pancreatic necrosis virus

doi:10.1006/fsim.2001.0397

Fish & Shellfish Immunology

Volume 13, Issue 3, September 2002, Pages 221-241

https://doi.org/10.1006/fsim.2001.0397 Get rights and content

Abstract

Type I interferons (IFN) establish an antiviral state in vertebrate cells by inducing expression of Mx and other antiviral proteins. We have studied the effect of Atlantic salmon interferon-like activity (AS-IFN) and poly I:C on the Mx protein expression and antiviral activity against infectious salmon anaemia virus (ISAV) and infectious pancreatic necrosis virus (IPNV) in the Atlantic salmon cell lines SHK-1 and TO. The double-stranded RNA poly I:C is an inducer of type I IFN in vertebrates. A cell cytotoxicity assay and measurements of virus yield were used to measure protection of cells against virus infection. Maximal induction of Mx protein in TO and SHK-1 cells occurred 48 h after poly I:C stimulation and 24 h after AS-IFN stimulation. TO cells pretreated with AS-IFN or poly I:C were protected from infection with IPNV 24 to 96 h after stimulation. Poly I:C or AS-IFN induced a minor protection against ISAV infection in SHK-1 cells, but no protection was induced against ISAV in TO cells. Western blot analysis showed that ISAV induced expression of Mx protein in TO and SHK-1 cells whereas IPNV did not induce Mx protein expression. These results suggest that ISAV and IPNV have very different sensitivities to IFN-induced antiviral activity and have developed different strategies to avoid the IFN-system of Atlantic salmon. Moreover, Atlantic salmon Mx protein appears not to inhibit replication of ISAV.

References (51)

P. Dobos
The molecular biology of infectious pancreatic necrosis virus (IPNV)
Annual Review of Fish Diseases
(1995)
C. Rogel-Gaillard et al.
In vitro induction of interferon-like activity from rainbow trout leucocytes stimulated by Egtvedt virus
Fish & Shellfish Immunology
(1993)
J. Snegaroff
Induction of interferon synthesis in rainbow-trout leukocytes by various homeotherm viruses
Fish & Shellfish Immunology
(1993)
R. Nygaard et al.
Induction of Mx-protein by interferon and double-stranded RNA in salmonid cells
Fish & Shellfish Immunology
(2000)
B. Robertsen et al.
Molecular cloning of double-stranded RNA inducible Mx genes from Atlantic salmon (Salmo salar L.)
Developmental and Comparative Immunology
(1997)
J.Y. Lee et al.
Cloning and analysis of expression of Mx cDNA in Japanese flounder, Paralichthys olivaceus
Developmental and Comparative Immunology
(2000)
V. Chieux et al.
Inhibition of coxsackievirus B4 replication in stably transfected cells expressing human MxA protein
Virology
(2001)
L.H. Wong et al.
Interferon-resistant human melanoma cells are deficient in ISGF3 components, STAT1, STAT2, and p48-ISGF3gamma
Journal of Biological Chemistry
(1997)
R.D. MacDonald et al.
Infectious pancreatic necrosis virus persistently infects chinook salmon embryo cells independent of interferon
Virology
(1979)
B. Schumacher et al.
The chicken Mx promoter contains an ISRE motif and confers interferon inducibility to a reporter gene in chick and monkey cells
Virology
(1994)

L. Bazzigher et al.

Mx genes show weaker primary response to virus than other interferon-regulated genes

Virology

(1992)

J.R. Hong et al.

Apoptosis precedes necrosis of fish cell line with infectious pancreatic necrosis virus infection

Virology

(1998)

M. Dorson et al.

Interferon synthesis in rainbow trout fry following infection with infectious pancreatic necrosis virus

Fish & Shellfish Immunology

(1992)

A. Garcia-Sastre et al.

Influenza A virus lacking the NS1 gene replicates in interferon-deficient systems

Virology

(1998)

K. Falk et al.

Characterization of infectious salmon anemia virus, an orthomyxo-like virus isolated from Atlantic salmon (Salmo salar L.)

Journal of Virology

(1997)

S. Mjaaland et al.

Genomic characterization of the virus causing infectious salmon anemia in Atlantic salmon (Salmo salar L.): an orthomyxo-like virus in a teleost

Journal of Virology

(1997)

B. Krossøy et al.

The putative polymerase sequence of infectious salmon anemia virus suggests a new genus within the Orthomyxoviridae

Journal of Virology

(1999)

G.R. Stark et al.

How cells respond to interferons

Annual Review of Biochemistry

(1998)

S. Goodbourn et al.

Interferons: cell signaling, immune modulation, antiviral responses and virus countermeasures

Journal of General Virology

(2000)

W.D. Eaton

Antiviral activity in 4 species of salmonids following exposure to poly inosinic–cytidylic acid

Diseases of Aquatic Organisms

(1990)

P. De Kinkelin et al.

Interferon production in rainbow trout (Salmo gairdneri) experimentally infected with Egtved virus

Journal of General Virology

(1973)

T. Renault et al.

Spectrophotometric method for titration of trout interferon, and its application to rainbow trout fry experimentally infected with viral hemaorrhagic septicemia virus

Diseases of Aquatic Organisms

(1991)

E. De Clercq

Interferon induction by polynucleotides, modified polynucleotides and polycarboxylates

Methods in Enzymology

(1981)

P. Boudinot et al.

vig-1, a new fish gene induced by the rhabdovirus glycoprotein, has a virus-induced homologue in humans and shares conserved motifs with the MoaA family

Journal of Virology

(1999)

Y. Grimholt et al.

The major histocompatibility complex in fish

Revue Scientifique et Technique

(1998)

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^f1: Corresponding author. E-mail: [email protected]

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Regular ArticlesEffect of double-stranded RNA and interferon on the antiviral activity of Atlantic salmon cells against infectious salmon anemia virus and infectious pancreatic necrosis virus

Abstract

Annual Review of Fish Diseases

Fish & Shellfish Immunology

Fish & Shellfish Immunology

Fish & Shellfish Immunology

Developmental and Comparative Immunology

Developmental and Comparative Immunology

Virology

Journal of Biological Chemistry

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Virology

Virology

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Fish & Shellfish Immunology

Virology

Characterization of infectious salmon anemia virus, an orthomyxo-like virus isolated from Atlantic salmon (Salmo salar L.)

Journal of Virology

Genomic characterization of the virus causing infectious salmon anemia in Atlantic salmon (Salmo salar L.): an orthomyxo-like virus in a teleost

Journal of Virology

The putative polymerase sequence of infectious salmon anemia virus suggests a new genus within the Orthomyxoviridae

Journal of Virology

How cells respond to interferons

Annual Review of Biochemistry

Interferons: cell signaling, immune modulation, antiviral responses and virus countermeasures

Journal of General Virology

Antiviral activity in 4 species of salmonids following exposure to poly inosinic–cytidylic acid

Diseases of Aquatic Organisms

Interferon production in rainbow trout (Salmo gairdneri) experimentally infected with Egtved virus

Journal of General Virology

Spectrophotometric method for titration of trout interferon, and its application to rainbow trout fry experimentally infected with viral hemaorrhagic septicemia virus

Diseases of Aquatic Organisms

Interferon induction by polynucleotides, modified polynucleotides and polycarboxylates

Methods in Enzymology

vig-1, a new fish gene induced by the rhabdovirus glycoprotein, has a virus-induced homologue in humans and shares conserved motifs with the MoaA family

Journal of Virology

The major histocompatibility complex in fish

Revue Scientifique et Technique

Regular Articles
Effect of double-stranded RNA and interferon on the antiviral activity of Atlantic salmon cells against infectious salmon anemia virus and infectious pancreatic necrosis virus