Unprocessed foot-and-mouth disease virus capsid precursor displays discontinuous epitopes involved in viral neutralization.
A foot-and-mouth disease virus (FMDV) cDNA cassette containing sequences encoding the capsid precursor P1, peptide 2A and a truncated 2B (abbreviated P1-2A) of type C FMDV, has been modified to generate the authentic amino terminus and the myristoylation signal. This construct has been used to produce a recombinant baculovirus (AcMM53) which, upon infection of Spodoptera frugiperda insect cells, expressed a recombinant P1-2A precursor with a high yield. This polyprotein reacted with neutralizing monoclonal antibodies (MAbs) that bind to continuous epitopes of the major antigenic site A (also termed site 1) of capsid protein VP1. Unexpectedly, it also reacted with neutralizing MAbs which define complex, discontinuous epitopes previously identified on FMDV particles. The reactivity of MAbs with P1-2A was quantitatively similar to their reactivity with intact virus and, in both cases, the reactivity with MAbs that recognized discontinuous epitopes was lost upon heat denaturation of the antigen. The finding that a capsid precursor may fold in such a way as to maintain discontinuous epitopes involved in virus neutralization present on the virion surface opens the possibility of using unprocessed capsid precursors as novel antiviral immunogens.
Sáiz
J C
JC
Centro de Biología Molecular Severo Ochoa, Consejo Superior de Investigaciones Científicas, Universidad Autónoma de Madrid, Spain.
Cairó
J
J
Medina
M
M
Zuidema
D
D
Abrams
C
C
Belsham
G J
GJ
Domingo
E
E
Vlak
J M
JM
D000818:Animals; D000911:Antibodies, Monoclonal; D005537:Aphthovirus / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D002213:Capsid / Q000276:immunology*; D002478:Cells, Cultured; D004279:DNA, Viral; D000939:Epitopes / Q000276:immunology*; D005814:Genes, Viral; D008969:Molecular Sequence Data; D009036:Moths; D009500:Neutralization Tests; D011498:Protein Precursors / Q000276:immunology*; D011995:Recombination, Genetic
D000911:Antibodies, Monoclonal; D004279:DNA, Viral; D000939:Epitopes; D011498:Protein Precursors