Neutralization of Egtved virus pathogenicity to cell cultures and fish by monoclonal antibodies to the viral G protein

Lorenzen, Niels; Olesen, Niels Jørgen; Jørgensen, P. E. V.

doi:10.1099/0022-1317-71-3-561

Cited by 121 publications

(116 citation statements)

References 33 publications

(24 reference statements)

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“…Neutralizing antibodies are directed exclusively against the G protein and are the most important component of the protective immune response against VHSV (Lorenzen et al, 1990). They exert their neutralizing activity, either at early stages of virus attachment to the cell membrane by blocking binding to the cellular receptor, or later in the cycle by inhibiting the fusion process.…”

Section: Introductionmentioning

confidence: 99%

Mutations in the glycoprotein of viral haemorrhagic septicaemia virus that affect virulence for fish and the pH threshold for membrane fusion.

Gaudin

Kinkelin

Benmansour

1999

Journal of General Virology

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To study the molecular basis of virulence of viral haemorrhagic septicaemia virus (VHSV), we used a cross-reactive neutralizing MAb to select MAb-resistant (MAR) mutants with reduced pathogenicity for fish. From sequence determination of the G gene of MAR mutants, attenuated laboratory variant and avirulent field strains, we identified two distant regions of the glycoprotein associated with virulence : region I (aa 135-161), homologous to the putative fusion peptide of both rabies virus (RV) and vesicular stomatitis virus (VSV), and region II (surrounding aa 431-433), homologous to RV and VSV domains controlling the conformational changes necessary for the fusion process to take place. Simultaneous mutations in both regions resulted in the most attenuated phenotype and we obtained genetic evidence that regions I and II may be structurally linked. As the MAR mutants had mutations in or near domains involved in fusion, the fusion properties of VHSV and its variants were analysed. This work allowed us to postulate that the fusion domain of VHSV is probably constituted of two distinct regions of the protein connected through a disulfide bridge between cysteines 110 and 152. Finally, we obtained evidence suggesting that the pH threshold for fusion is a determinant for virulence : restriction of fusion to a more acidic pH was associated with attenuation for the variant tr25 which had a shift of the threshold for maximal fusion from pH 6n30 (for the parental strain) to pH 6n00 ; conversely, two field strains which had maximal fusion at pH 6n60 were the most virulent.

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Section: Introductionmentioning

confidence: 99%

Mutations in the glycoprotein of viral haemorrhagic septicaemia virus that affect virulence for fish and the pH threshold for membrane fusion.

Gaudin

Kinkelin

Benmansour

1999

Journal of General Virology

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“…Cell pellets were dissolved by boiling in reducing sample buffer for SDS-PAGE. SDS-PAGE and irnmunoblotting were performed as before (Lorenzen et al 1990). Purified virus was u.sed a positive control antigen.…”

Section: Methodsmentioning

confidence: 99%

“…Because neutralizing ability of a n antibody to a rhabdovirus (i.e. VHSV) in cell culture does not always correlate with its protection properties in vivo (LeFrancois 1984) and nonneutralizing anti-VHSV antibodies can be VHS protective when injected in vivo (Lorenzen et al 1990), the present work is also an attempt to identify some of the major antigenic sites (neutralizing or not neutralizing) that can be mimicked by synthetic peptides in the gpG of VHSV. Mapping of the linear antibody epitopes in the gpG could either give information concerning the failure of recombinant gpG as a vaccine and/or identify regions of the protein which could be important to include in a future vaccine.…”

Section: Introductionmentioning

confidence: 99%

“…Sequencing of MAb neutralization resistant (MAR) mutants of rabies (Benmansour et al 1991, Raux et al 1995, VSV (Vandepol et al 1986, Luo et al 1988, IHNV (Kim et al 1994) and VHSV (Bearzotti et al 1995) has suggested that 2 regions being distant in the primary structure of the gpG might be associated in the native conformational stage of the protein. Only a few linear reduction-resistant neutralization epitopes have been identified in rabies (VanDeerHeijden et al 1993, Ni et al 1995, VSV (LeFrancois & Lyles 1982, Volk et al 1982, Keil & Wagner 1989, Grigera et al 1992 and IHNV (Leong et al 1995) but none yet in VHSV (Lorenzen et al 1990, Sanz & Col1 1992. The present study focusses on mapping of linear antibody epitopes on the gpG of VHSV.…”

Section: Introductionmentioning

confidence: 99%

“…Reduction of the disulfide bonds of the gpG inhibited the binding of many neutralizing monoclonal antibodies (MAbs) to rhabdoviruses such as rabies (Benmansour et al 1991), vesicular stomatitis virus, VSV (Keil & Wagner 1989), VHSV (Lorenzen et al 1990, Bearzotti et al 1995 and infectious haematopoietic necrosis virus, IHNV (Kim et al 1994, Huang et al 1996. Sequencing of MAb neutralization resistant (MAR) mutants of rabies (Benmansour et al 1991, Raux et al 1995, VSV (Vandepol et al 1986, Luo et al 1988, IHNV (Kim et al 1994) and VHSV (Bearzotti et al 1995) has suggested that 2 regions being distant in the primary structure of the gpG might be associated in the native conformational stage of the protein.…”

Section: Introductionmentioning

confidence: 99%

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Mapping of linear antibody epitopes of the glycoprotein of VHSV, a salmonid rhabdovirus

Fernandez‐Alonso¹,

Lorenzo²,

Pérez³

et al. 1998

Dis. Aquat. Org.

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Antibody linear epitopes of the glycoprotein G (gpG) of the viral haemorrhagic septicaemia virus (VHSV), a rhabdovirus of salrnonids, were mapped by pepscan using overlapping 15-mer peptides covering the entire gpG sequence and ELISA with polyclonal and monoclonal murine and polyclonal trout antibodies. Among the regions recognized in the pepscan by the polyclonal antibodies (PAbs) were the previously identified phosphatidylserine binding heptad-repeats (Estepa & Col1 1996; Virology 216:60-70) and leucocyte stimulating peptides (Lorenzo et al. 1995; Virology 212:348-355). Among 17 monoclonal antibodies (MAbs), only 2 non-neutralizing MAbs, 110 (aa 139-153) and IPlH3 (aa 399-413), could b e mapped to specific peptides in the pepscan of the gpG. Mapping of these MAbs was confirmed by immunoblotting with recombinant proteins and/or other synthetic peptides covering those sequences.None of the neutralizing MAbs tested reacted with any of the gpG peptides. Previously mapped MAb resistant mutants in the gpG did not coincide with any of the linear epitopes defined by the pepscan strategy, suggesting the complementarity of the 2 methods for the identification of antibody recogn~tion sites.

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Vaccination against Viral Hemorrhagic Septicemia and Infectious Hematopoietic Necrosis

Biacchesi

Brémont

2014

Fish Vaccination

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Neutralization of Egtved virus pathogenicity to cell cultures and fish by monoclonal antibodies to the viral G protein

Cited by 121 publications

References 33 publications

Mutations in the glycoprotein of viral haemorrhagic septicaemia virus that affect virulence for fish and the pH threshold for membrane fusion.

Mutations in the glycoprotein of viral haemorrhagic septicaemia virus that affect virulence for fish and the pH threshold for membrane fusion.

Mapping of linear antibody epitopes of the glycoprotein of VHSV, a salmonid rhabdovirus

Vaccination against Viral Hemorrhagic Septicemia and Infectious Hematopoietic Necrosis

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