Mapping the neutralizing epitopes on the glycoprotein of infectious haematopoietic necrosis virus, a fish rhabdovirus.
Twelve neutralizing monoclonal antibodies (MAbs) against the fish rhabdovirus, infectious haematopoietic necrosis virus (IHNV), were used to select 20 MAb escape mutants. The nucleotide sequence of the entire glycoprotein (G) gene was determined for six mutants representing differing cross-neutralization patterns and each had a single nucleotide change leading to a single amino acid substitution within one of three regions of the protein. These data were used to design nested PCR primers to amplify portions of the G gene of the 14 remaining mutants. When the PCR products from these mutants were sequenced, they also had single nucleotide substitutions coding for amino acid substitutions at the same, or nearby, locations. Of the 20 mutants for which all or part of the glycoprotein gene was sequenced, two MAbs selected mutants with substitutions at amino acids 230-231 (antigenic site I) and the remaining MAbs selected mutants with substitutions at amino acids 272-276 (antigenic site II). Two MAbs that selected mutants mapping to amino acids 272-276, selected other mutants that mapped to amino acids 78-81, raising the possibility that this portion of the N terminus of the protein was part of a discontinuous epitope defining antigenic site II. CLUSTAL alignment of the glycoproteins of rabies virus, vesicular stomatitis virus and IHNV revealed similarities in the location of the neutralizing epitopes and a high degree of conservation among cysteine residues, indicating that the glycoproteins of three different genera of animal rhabdoviruses may share a similar three-dimensional structure in spite of extensive sequence divergence.
Huang
C
C
School of Fisheries, University of Washington, Seattle 98195, USA.
Chien
M S
MS
Landolt
M
M
Batts
W
W
Winton
J
J
D000595:Amino Acid Sequence; D000818:Animals; D000956:Antigens, Viral / Q000235:genetics* / Q000276:immunology; D001483:Base Sequence; D002460:Cell Line; D004279:DNA, Viral; D018604:Epitope Mapping; D000939:Epitopes / Q000235:genetics*; D005399:Fishes / Q000821:virology*; D006023:Glycoproteins / Q000235:genetics* / Q000276:immunology; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D012209:Rhabdoviridae / Q000235:genetics / Q000276:immunology*; D017386:Sequence Homology, Amino Acid
D000956:Antigens, Viral; D004279:DNA, Viral; D000939:Epitopes; D006023:Glycoproteins