Cloning of the rainbow trout (Oncorhynchus mykiss) Mx2 and Mx3 cDNAs and characterization of trout Mx protein expression in salmon cells

Trobridge, Grant D.; Chiou, Pinwen Peter; Leong, J. A. C.

doi:10.1128/jvi.71.7.5304-5311.1997

Cited by 152 publications

(28 citation statements)

References 35 publications

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“…Multiple lines of evidence suggest that if the IFN system is induced prior to IHNV challenge, the animals are protected against challenge (Eaton 1990;Ooi, Verjan, Haraguchi, Oshima, Kondo, Hirono, Aoki, Kiyono & Yuki 2008). Infection with IHNV rapidly induces a strong host innate IFN response (Trobridge, Chiou & Leong 1997b;Trobridge et al 1997a;Purcell et al 2004) but, in some cases, the virus can continue to replicate in the presence of this response. In the study presented here, U-type virus replicated and caused mortality despite the IFN response evident by 3 days p.i.…”

Section: Discussionmentioning

confidence: 99%

Infectious haematopoietic necrosis virus genogroup‐specific virulence mechanisms in sockeye salmon, Oncorhynchus nerka (Walbaum), from Redfish Lake, Idaho

Purcell

Garver²,

Conway

et al. 2009

Journal of Fish Diseases

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Characterization of infectious haematopoietic necrosis virus (IHNV) field isolates from North America has established three main genogroups (U, M and L) that differ in host-specific virulence. In sockeye salmon, Oncorhynchus nerka, the U genogroup is highly virulent, whereas the M genogroup is nearly non-pathogenic. In this study, we sought to characterize the virus-host dynamics that contribute to genogroup-specific virulence in a captive stock of sockeye salmon from Redfish Lake in Idaho. Juvenile sockeye salmon were challenged by immersion and injection with either a representative U or M viral strain and sampled periodically until 14 days post-infection (p.i.). Fish challenged with each strain had positive viral titre by day 3, regardless of challenge route, but the fish exposed to the M genogroup virus had significantly lower virus titres than fish exposed to the U genogroup virus. Gene expression analysis by quantitative reverse transcriptase PCR was used to simultaneously assess viral load and host interferon (IFN) response in the anterior kidney. Viral load was significantly higher in the U-challenged fish relative to M-challenged fish. Both viruses induced expression of the IFN-stimulated genes (ISGs), but expression was usually significantly lower in the M-challenged group, particularly at later time points (7 and 14 days p.i.). However, ISG expression was comparable with 3 days post-immersion challenge despite a significant difference in viral load. Our data indicated that the M genogroup virus entered the host, replicated and spread in the sockeye salmon tissues, but to a lesser extent than the U genogroup. Both virus types induced a host IFN response, but the high virulence strain (U) continued to replicate in the presence of this response, whereas the low virulence strain (M) was cleared below detectable levels. We hypothesize that high virulence is associated with early in vivo replication allowing the virus to achieve a threshold level, which the host innate immune system cannot control.

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Section: Discussionmentioning

confidence: 99%

Infectious haematopoietic necrosis virus genogroup‐specific virulence mechanisms in sockeye salmon, Oncorhynchus nerka (Walbaum), from Redfish Lake, Idaho

Purcell

Garver²,

Conway

et al. 2009

Journal of Fish Diseases

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“…Primary antibodies used in this study were the monoclonal antibodies against VP2 (␣VP2, 1:1000 dilution) (kindly provided by K.E. Christie, Intervet Norbio), the polyclonal antibodies against VP1 (␣VP1, 1:1000 dilution), VP3 (␣VP3, 1:4000 dilution) (produced as described in (Pedersen et al, 2007), a polyclonal Mx antibody (␣-Mx, 1:1000 dilution) (Trobridge et al, 1997) and the polyclonal eEF2 antibody (␣-eEF2, 1:1000 dilution (Hansen et al, 2008) (Cell Signaling Technology). Goat anti-rabbithorseradish peroxidase (HRP) antibody or goat anti-mouse-HRP antibody (Santa Cruz Biotechnology) diluted 1:25,000 were used as secondary antibodies.…”

Section: Gel Electrophoresis Western Blotting and Antibodiesmentioning

confidence: 99%

The interplay between infectious pancreatic necrosis virus (IPNV) and the IFN system: IFN signaling is inhibited by IPNV infection

et al. 2009

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Infectious pancreatic necrosis virus (IPNV) is a major pathogen in the aquaculture industry worldwide. Factors contributing to IPNV pathogenicity are yet poorly understood. Indications of IPNV being able to evade or counteract innate host defense come from its lack of ability to induce strong type I interferon (IFN) responses in cell culture. We show here that addition of salmon rIFN-alpha1 to cells prior to IPNV infection halts the viral protein synthesis and prevents processing of pVP2 into mature VP2. Furthermore, compared to pre-treatment with IFN-alpha1 the antiviral state in cells infected with IPNV prior to IFN-treatment, was antagonized by IPNV, as detected by higher viral titers, faster viral protein synthesis and also by reduced Mx expression. The longer headstart the virus gets, the more prominent is the weakening of IFN signaling. IPNV VP4 and VP5 inhibit IFN-induced expression from the Mx promoter, indicating that these proteins contribute to the antagonistic effect.

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“…The antiserum was generated against a fragment of the rainbow trout Mx3 protein as described. (24) The plasmid used to express the rainbow trout Mx3 fragment was a gift from Jo-Ann C. Leong (Department of Microbiology, Oregon State University, Corvallis, OR). Horseradish peroxidase (HRP)-conjugated goat antirabbit antibody (Santa Cruz Biotechnology, Inc., Santa Cruz, CA) diluted 1:10,000 was used as secondary antibody.…”

Section: Immunoblot Analysismentioning

confidence: 99%

Atlantic Salmon Interferon Genes: Cloning, Sequence Analysis, Expression, and Biological Activity

Robertsen

Bergan²,

Røkenes³

et al. 2003

Journal of Interferon & Cytokine Research

236

175

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In this work, we report cDNA cloning of two type I interferons (IFNs) from the head kidney of Atlantic salmon, called SasaIFN-alpha1 (829 bp) and SasaIFN-alpha2 (1290 bp). Both translate into 175 amino acid precursor molecules showing 95% amino acid sequence identity. The precursors have a putative 23 amino acid signal peptide, which suggests that the mature Atlantic salmon IFNs contain 152 amino acids (18.2 kDa). Salmon IFN appears to have five alpha-helices, similar to mammalian and avian type I IFNs, and showed 45% sequence identity with zebrafish IFN, up to 29% identity with mammalian IFN-alpha sequences, and 17%-18% sequence identity with mammalian IFN-beta and chicken type I IFNs. Human embryonic kidney 293 cells transfected with the SasaIFN-alpha1 cDNA gene produced high titers of acid-stable antiviral activity, which protected salmonid cells against infectious pancreatic necrosis virus (IPNV) and also induced Mx protein in the cells. Poly(I)-poly(C) induced two IFN transcripts in head kidney of Atlantic salmon. Genomic IFN sequences contained four introns and five exons, which is different from the intronless type I IFN genes of birds and mammals.

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Cloning of the rainbow trout (Oncorhynchus mykiss) Mx2 and Mx3 cDNAs and characterization of trout Mx protein expression in salmon cells

Cited by 152 publications

References 35 publications

Infectious haematopoietic necrosis virus genogroup‐specific virulence mechanisms in sockeye salmon, Oncorhynchus nerka (Walbaum), from Redfish Lake, Idaho

Infectious haematopoietic necrosis virus genogroup‐specific virulence mechanisms in sockeye salmon, Oncorhynchus nerka (Walbaum), from Redfish Lake, Idaho

The interplay between infectious pancreatic necrosis virus (IPNV) and the IFN system: IFN signaling is inhibited by IPNV infection

Atlantic Salmon Interferon Genes: Cloning, Sequence Analysis, Expression, and Biological Activity

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