The free nucleocapsids of the viral haemorrhagic septicaemia virus contain two antigenically related nucleoproteins

Basurco, B.; Sanz, Fernando V. Díez; Marcotegui, M. A.; Coll, Julio

doi:10.1007/bf01310666

Cited by 21 publications

(18 citation statements)

References 16 publications

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“…The VHSV was grown in epithelial papullosum cyprini (EPC) cells and concentrated by polyethyleneglycol to about 70% purity as described before (25). The VHSV assay was similar to the microneutralization assay described before (26).…”

Section: Methodsmentioning

confidence: 99%

A Protein G Fragment from the Salmonid Viral Hemorrhagic Septicemia Rhabdovirus Induces Cell-to-Cell Fusion and Membrane Phosphatidylserine Translocation at Low pH

Estepa

Rocha

Más

et al. 2001

Journal of Biological Chemistry

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The fusion-related properties of segments p9, p3, p4, and p9 ؉ p2 surrounding the p2 phospholipid-binding domain of the protein G (pG) of the salmonid rhabdovirus of viral hemorrhagic septicemia (VHS) (Nuñ ez, E., Fernandez, A. M., Estepa, A., Gonzalez-Ros, J. M., Gavilanes, F., and Coll, J. M. (1998) Virology 243, 322-330; Estepa, A., and Coll, J. M. (1996) Virology 216, 60 -70), have been studied at neutral and fusion (low) pH values by using its derived peptides. Cell-to-cell fusion, translocation of phosphatidylserine, and inhibition of fusion of pG-transfected cells defined the p9 ؉ p2 (fragment 11, sequence 56 -110) as a fragment with higher specific activity for anionic phospholipid aggregation than the previously reported p2. While fragment 11, p2, and p3 showed interactions with anionic phospholipids, p9 and p4 showed no interactions with any phospholipids. When added to a cell monolayer model at low pH, fragment 11 induced pH-dependent cell-to-cell fusion and translocated phosphatidylserine from the inner to the outer leaflet of the membrane. At low pH and in the presence of anionic phospholipids, fragment 11 showed more than 80% ␤-sheet conformation (IR and CD spectroscopies). Finally, anti-fragment 11 antibodies inhibited low pH-dependent pG-transfected cell-to-cell fusion. All of the data support the conclusion that fragment 11 is a primary determinant of some of the viral cell fusion events in VHSV.

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Section: Methodsmentioning

confidence: 99%

A Protein G Fragment from the Salmonid Viral Hemorrhagic Septicemia Rhabdovirus Induces Cell-to-Cell Fusion and Membrane Phosphatidylserine Translocation at Low pH

Estepa

Rocha

Más

et al. 2001

Journal of Biological Chemistry

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“…The VHSV 07.71, isolated in France from rainbow trout Oncorynchus mykiss (LeBerre et al 1977), was grown and/or assayed for infectivity in epithelioma papillosum cyprini (EPC) cells (DeKinkelin 1972) and purified as described previously (Basurco et al 1991). Anti-p2 and anti-Gs polyclonal antibodies (PAbs) were obtained as described previously (Estepa & Coll 1996).…”

Section: Methodsmentioning

confidence: 99%

Temperature and pH requirements for viral haemorrhagic septicemia virus induced cell fusion

Estepa¹,

Coll²

1997

Dis. Aquat. Org.

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The rhabdovirus causlng v~r a l haemorrhaglc septlcema (VHS), an Important disease of salmonlds, Induced forrnatlon of large syncytia in fish cell monolayers by cell fusion For maximal percentage of fus~on, not only was a pH of 5 6 needed but also a temperature of at least 14°C The fusion was dependent on the G proteln of VHSV as shown by inhibition of fuslon with speclflc anti-G polyclonal and monoclonal antibodies Part of the region of the G proteln Involved In fusion was mapped to amino acids 82 to 109 because anti-peptide antibodies blndlng to that reglon also inhlblted fuslon The fusion assay descnbed here can b e useful for studying the functionality of the G proteln of fish rhabdovlruses in the early steps of Infection and thls information can In turn, help to designing effectwe vaccination and/or therapeutic strategies against fish rhabdovlruses The descnbed fusion assay could also be a p p l~e d to other flsh rhabdoviruses such as infect~ous haematopoletic necrosls virus (IHNV) and spnng viraemia of carp vlrus (SVCV) KEY WORDS Viral haemorrhagic septicaemia Rhabdovirus Neutrahzing-enhancing a n t~b o d y Monoclonal antibodies Fusion

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“…Visceral homogenates may nosis in some cases (Basurco et al 1991). Viral protein more effectively identify virus-carriers, but it is not content is a measure of the number of virions; 1 n g of practical to kill a large number of fish.…”

Section: Antibodies To Viral Proteinsmentioning

confidence: 99%

Techniques for diagnosing viral diseases of salmonid fish

Sanz¹,

Coll²

1992

Dis. Aquat. Org.

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Cell culture for amplification and the techniques most used for identification of salmonid viruses -neutralization, immunofluorescence and, to a lesser extent, immunoperoxldase, complen~ent fixation, agglutination, electron microscopy, immunodifussion or radioimmunoassay -may soon b e replaced by other techniques such as enzyme immunoassays (immunodot and enzyme-linked immunosorbent assay, ELISA) and hybridization with DNA probes. It is expected that developments in monoclonal antibodies (MAbs) and amplification by the polymerase chain reaction (PCR) will increase sensitivity of enzyme immunoassays and DNA hybridizations, respectively. Some of these new methods should provide detection of the low levels of virus present in adult carriers and perhaps in eggs (although this is more complicated). Other &agnostic methods, such as measurement of virus-specific salmonid immunoglobulins (Igs) by ELISA or stimulation of immunological cellular memory by in vitro CO-culture of salmonid lymphocytes with viral proteins, could also b e further developed.

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The free nucleocapsids of the viral haemorrhagic septicaemia virus contain two antigenically related nucleoproteins

Cited by 21 publications

References 16 publications

A Protein G Fragment from the Salmonid Viral Hemorrhagic Septicemia Rhabdovirus Induces Cell-to-Cell Fusion and Membrane Phosphatidylserine Translocation at Low pH

A Protein G Fragment from the Salmonid Viral Hemorrhagic Septicemia Rhabdovirus Induces Cell-to-Cell Fusion and Membrane Phosphatidylserine Translocation at Low pH

Temperature and pH requirements for viral haemorrhagic septicemia virus induced cell fusion

Techniques for diagnosing viral diseases of salmonid fish

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