Characterisation of the syncytia formed by VHS salmonid rhabdovirus G gene transfected cells.
Protein G expression and cell-to-cell fusion of cells transfected with the G gene of viral haemorrhagic septicaemia virus (VHSV) has been characterised. The presence of protein G in the membrane of transfected cells was confirmed by staining with Abs (Abs) and FACS. The subsequent formation of syncytia by membrane fusion of transfected cells required transfection with a wild type G gene and a low pH step. Mice Abs made against the protein G regions involved in fusion and neutralising monoclonal Abs (MAbs) as well as MAbs against some linear epitopes inhibited syncytia formation, thus confirming that syncytia formation was G-dependent. Similarly, Abs from trout immunised with purified VHSV or protein G inhibited syncytia formation whereas Abs from non-immunised or non-infected animals did not. Abs from mice or trout with the highest neutralisation titres also showed the highest percentage of inhibition of syncytia. While the main utility of these observations might be to further the understanding of the complex trout antibody response against VHSV and in the follow up of VHSV immunisation attempts, they may also have some future diagnostic potential for countries were work with VHSV is not allowed.
Rocha
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A
SGIT, INIA, Biotecnología, Crt. La Coruña, Km. 7, 28040 Madrid, Spain.
Ruiz
S
S
Tafalla
C
C
Coll
J M
JM
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics; D005260:Female; D005393:Fish Diseases / Q000276:immunology / Q000821:virology*; D015726:Giant Cells / Q000276:immunology* / Q000821:virology; D031941:Hemorrhagic Septicemia, Viral / Q000276:immunology* / Q000821:virology; D006863:Hydrogen-Ion Concentration; D008561:Membrane Fusion / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D016297:Mutagenesis, Site-Directed; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000276:immunology*; D009500:Neutralization Tests / Q000662:veterinary; D029141:Novirhabdovirus / Q000235:genetics / Q000276:immunology*; D014162:Transfection / Q000662:veterinary; D014337:Trout / Q000276:immunology*; D001616:beta-Galactosidase / Q000276:immunology
D000914:Antibodies, Viral; D004279:DNA, Viral; C029152:G-substrate; D009419:Nerve Tissue Proteins; D001616:beta-Galactosidase