Epitope mapping and characterization of the infectious hematopoietic necrosis virus glycoprotein, using fusion proteins synthesized in Escherichia coli

Xu, Long; Mourich, Dan V.; Engelking, H. Mark; Ristow, Sandra S.; Arnzen, Jeanene M.; Leong, Jo‐Ann C.

doi:10.1128/jvi.65.3.1611-1615.1991

Cited by 45 publications

(9 citation statements)

References 19 publications

(17 reference statements)

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“…At amino acid sites 247, 256, and 270, the U and L genogroup viruses contained an arginine, lysine, and glutamic acid, respectively, while the three M genogroup viruses had a glutamine, a glycine or glutamine, and a glycine or valine, respectively. These sites do not coincide with previously identified antigenic sites (Xu et al 1991;Huang et al 1994Huang et al , 1996Kim et al 1994) or potential N-glycosylation sites. The genogroup-specific change at amino acid site 247 correlates with a change at nucleotide 786 of the G gene, which was shown previously to consistently differ between U and M genogroup viruses in an alignment of partial G sequences from 30 IHNV isolates (17 U genogroup viruses and 13 M genogroup viruses; Garver et al 2003).…”

Section: Virulence Of Ihnv Genogroupscontrasting

confidence: 75%

Virulence Comparisons of Infectious Hematopoietic Necrosis Virus U and M Genogroups in Sockeye Salmon and Rainbow Trout

2006

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Infectious hematopoietic necrosis virus (IHNV) is an aquatic rhabdovirus that infects salmonids in the Pacific Northwest of the United States, Europe, and Asia. Isolates of IHNV have been phylogenetically classified into three major viral genogroups, designated U, M, and L. To characterize virulence of IHNV in the context of these three viral genogroups, seven strains of IHNV (three U genogroup strains, three M strains, and one L strain) were compared for their pathogenicity in juvenile sockeye salmon Oncorhynchus nerka, kokanee (lacustrine sockeye salmon), and rainbow trout O. mykiss. Fish were waterborne-exposed to the different viral strains, and virulence was assessed by comparing mortality curves and final cumulative percent mortality (CPM) in both species of fish at 10°C and 15°C. In sockeye salmon and kokanee, the U genogroup virus types were extremely virulent, causing average CPMs of 69-100%, while the M genogroup virus types caused very little or no mortality (CPM = 0-4%). The endangered Redfish Lake sockeye salmon stock exhibited extreme differences in susceptibility to the U and M genogroups. Conversely, in two stocks of rainbow trout, the M genogroup virus types were more virulent, inducing average CPMs of 25-85%, while the U genogroup viruses caused lower mortality (CPM = 5-41%). In both fish species, the single L genogroup strain caused low to intermediate mortality (CPM = 13-53%). Viral glycoprotein sequence comparisons of the seven challenge strains revealed three amino acid sites (247, 256, and 270) that consistently differed between the U and M genogroups, possibly contributing to pathogenicity differences.

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Section: Virulence Of Ihnv Genogroupscontrasting

confidence: 75%

Virulence Comparisons of Infectious Hematopoietic Necrosis Virus U and M Genogroups in Sockeye Salmon and Rainbow Trout

2006

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“…The key viral protein appears to be the glycoprotein in both IHNV and VHSV. Studies have shown that recombinant IHNV G protein fragments are capable of providing protection against the virus (15,46). When fragments of the gene were cloned and expressed in E. coli, effective protection was obtained, particularly with Cterminal portions of the G protein.…”

Section: Discussionmentioning

confidence: 99%

“…Cloning and nucleotide sequencing of cDNAs of IHNV and VHSV glycoprotein genes have been described (18,35). Studies have shown that portions of the IHNV G protein expressed in Escherichia coli are protective for rainbow trout and that the VHSV G protein is immunogenic in rainbow trout (22,46).…”

mentioning

confidence: 99%

Recombinant infectious hematopoietic necrosis virus and viral hemorrhagic septicemia virus glycoprotein epitopes expressed in Aeromonas salmonicida induce protective immunity in rainbow trout (Oncorhynchus mykiss)

Noonan

Enzmann

Trust

1995

Appl Environ Microbiol

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Fragments of the glycoprotein genes of viral hemorrhagic septicemia virus (VHSV) and infectious hematopoietic necrosis virus (IHNV) were cloned into a bacterial broad-host-range expression vector under the control of the plac promoter. Western blot (immunoblot) analysis with monoclonal antibodies specific to the glycoproteins demonstrated the inducible expression of the fusion proteins in Escherichia coli. Aeromonas salmonicida is the causative agent of furunculosis in salmonid fish. It was confirmed that an avirulent strain of A. salmonicida, A440, which contains a deletion in the structural gene for the paracrystalline surface protein array, will provide protective immunity against furunculosis when used as a live attenuated vaccine. The plasmid-encoded viral epitopes were then mobilized into A440 for use as a shuttle system for the expression of fragments of the glycoprotein genes of IHNV and VHSV. Vaccination of rainbow trout with A440 containing the viral epitopes resulted in the development of protective immunity against both VHSV and IHNV. This indicates that the use of cloned fragments of the glycoproteins and the use of A. salmonicida as a shuttle system constitute a feasible approach to fish vaccine development.

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“…The membrane was then incubated for 1 h at room temperature with 136J monoclonal antibody (mAb) in TBST. 136J is a mAb raised against the 67‐kDa glycoprotein of IHNV (Xu, Mourich, Engelking, Ristow, Arnzen & Leong 1991). The membrane was subsequently washed three times for 10 min each with TBST and then incubated for 1 h with goat anti‐mouse secondary antibody conjugated to horseradish peroxidase (Bio‐Rad) at a 1:3000 dilution in TBS.…”

Section: Methodsmentioning

confidence: 99%

Inhibition of infectious haematopoietic necrosis virus in cell cultures with peptide‐conjugated morpholino oligomers

Alonso

Stein

Thomann

et al. 2005

Journal of Fish Diseases

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Delivery of phosphorodiamidate morpholino oligomers (PMO) into fish cells in vitro and tissues in vivo was examined. Uptake was evaluated by fluorescence microscopy and flow cytometry after treating cultured cells or live rainbow trout with 3' fluorescein-tagged PMO. Arginine-rich peptide conjugated to the 5' end of the PMO markedly enhanced cellular uptake in culture by 8- to 20-fold compared with non-peptide-conjugated PMO as determined by flow cytometry. Enhanced uptake of PMO conjugated to peptide was also observed in tissues of fish treated by immersion. The efficacy of PMO as inhibitors of infectious haematopoietic necrosis virus (IHNV) replication was determined in vitro. Peptide-conjugated PMOs targeting sequences within the IHNV genomic RNA (negative polarity) or antigenomic RNA (positive polarity) significantly inhibited replication in a dose-dependent and sequence-specific manner. A PMO complementary to sequence near the 5' end of IHNV genomic RNA was the most effective, diminishing titre by 97%, as measured by plaque assay and Western blot. These data demonstrate that replication of a negative-stranded non-segmented RNA virus can be inhibited by antisense compounds that target positive polarity viral RNA, or by a compound that targets negative polarity viral RNA.

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Epitope mapping and characterization of the infectious hematopoietic necrosis virus glycoprotein, using fusion proteins synthesized in Escherichia coli

Cited by 45 publications

References 19 publications

Virulence Comparisons of Infectious Hematopoietic Necrosis Virus U and M Genogroups in Sockeye Salmon and Rainbow Trout

Virulence Comparisons of Infectious Hematopoietic Necrosis Virus U and M Genogroups in Sockeye Salmon and Rainbow Trout

Recombinant infectious hematopoietic necrosis virus and viral hemorrhagic septicemia virus glycoprotein epitopes expressed in Aeromonas salmonicida induce protective immunity in rainbow trout (Oncorhynchus mykiss)

Inhibition of infectious haematopoietic necrosis virus in cell cultures with peptide‐conjugated morpholino oligomers

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