Neutralizing epitopes of type O foot-and-mouth disease virus. II. Mapping three conformational sites with synthetic peptide reagents.
Four neutralizing monoclonal antibodies (MAbs), recognizing three functionally independent, conformational sites on type O foot-and-mouth disease virus (FMDV) failed to react with immobilized structural proteins or synthetic peptides but bound to the isolated capsid protein VP1 and peptides in solution. Inhibition ELISA techniques were, therefore, applied using peptide antigens and anti-peptide sera to block MAb binding to virus particles, permitting the identification of those portions of the VP1 protein contributing to the epitopes. The binding site of one MAb, which neutralized a range of type O FMDV isolates, was shown to have components within regions 146 to 150 and 200 to 213 of VP1 with a critical involvement of the amino acids at positions 146 and 206 or 207. The determinants recognized by two other MAbs which were directed at similar, but not identical, epitopes from a second site included components from the 200 to 213 and 143 to 146 regions with amino acids 143 and 144, respectively, appearing critical for the inhibition of the virus binding of the two antibodies. These results demonstrate that the two previously identified immunogenic tracts of VP1 are brought into proximity in the quaternary structure of the virion to form an antigenic domain containing several conformational epitopes, some of which are functionally independent. A fourth, strain-specific MAb was effectively blocked from reacting with virus by peptides corresponding to residues 161 to 180 and 200 to 213.
Parry
N R
NR
Department of Virology, Wellcome Biotechnology Ltd, Beckenham, Kent, U.K.
Barnett
P V
PV
Ouldridge
E J
EJ
Rowlands
D J
DJ
Brown
F
F
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification; D000956:Antigens, Viral / Q000032:analysis / Q000235:genetics / Q000276:immunology*; D005537:Aphthovirus / Q000235:genetics / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis / Q000235:genetics / Q000276:immunology*; D007106:Immune Sera / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification; D009500:Neutralization Tests; D010449:Peptide Mapping; D010455:Peptides / Q000276:immunology; D014764:Viral Proteins / Q000276:immunology*; D015678:Viral Structural Proteins
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D000939:Epitopes; D007106:Immune Sera; D010455:Peptides; D014764:Viral Proteins; D015678:Viral Structural Proteins