Characterization of a major neutralization domain of Ross river virus using anti-viral and anti-peptide antibodies.
The E2 glycoprotein of the alphavirus Ross River virus (RRV) contains three defined neutralization epitopes (a, b1 and b2) with determinants located between amino acids 216 and 251 in the linear sequence (Vrati et al., 1988, Virology 162, 346-353). The antigenic structure of this region has been examined using hyperimmune mouse antiserum against RRV and antiserum against four synthetic peptides representing linear amino acid sequences in the neutralization region of E2. In plaque reduction neutralization tests using hyperimmune antiserum to RRV, an RRV mutant altered at all three neutralization epitopes was markedly more resistant than the parental virus; variants altered at single epitopes could not be distinguished in these tests. Sera from mice immunized with synthetic RRV E2 peptides conjugated to keyhole limpet haemocyanin reacted, in a direct ELISA, with the specific region of RRV represented by the peptide. The same sera did not neutralize or immunoprecipitate RRV in solution or bind to RRV in a capture ELISA. The RRV peptides did not prime mice to react to a subimmunogenic dose of RRV; they did not bind monoclonal or polyclonal antibodies to RRV. We conclude that a significant proportion of the neutralizing antibody response in mice is elicited by epitopes a, b1, and b2 of RRV E2 and that the sites to which neutralizing antibodies bind are formed by complex folding.
Biochemistry Department, Faculty of Science, Australian National University, Canberra, ACT.
P J
PJ
Kerr
S
S
Fitzgerald
G W
GW
Tregear
L
L
Dalgarno
R C
RC
Weir
D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D007106:Immune Sera / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests; D010446:Peptide Fragments / Q000276:immunology; D011233:Precipitin Tests; D012398:Ross River virus / Q000276:immunology*; D014036:Togaviridae Infections / Q000517:prevention & control; D010948:Viral Plaque Assay; D014764:Viral Proteins / Q000276:immunology*
D000914:Antibodies, Viral; D000939:Epitopes; D007106:Immune Sera; D010446:Peptide Fragments; D014764:Viral Proteins