Mapping of linear epitopes on the capsid proteins of swine vesicular disease virus using monoclonal antibodies.
The antigenic linear map of swine vesicular disease virus (SVDV) has been studied using a repertoire of monoclonal antibodies (mAbs) raised against a recombinant SVDV polyprotein, P1. Peptide-scanning analyses, cross-reactivity studies with homologous and heterologous viruses and predicted location on a computer-generated three-dimensional model of the capsid proteins have allowed the identification of five main linear sites. Two sites, the N terminus of VP3 and amino acids 51-60 on VP1, correspond to internal areas, conserved not only between SVDV isolates but also in the related enterovirus coxsackievirus B5. In contrast, three other regions, amino acids 142-161 of VP2, 61-70 of VP3 and the C terminus of VP1, are exposed on the external face of the capsid and subjected to antigenic variation, even among different SVDV isolates. Further minor sites that were antigenically conserved were identified on VP4. In contrast with conformational sites described previously, none of the linear epitopes identified in this work is involved in neutralization of virus infectivity and post-infection swine sera did not inhibit the binding of mAbs with the relevant epitopes. Both of these observations suggest that linear epitopes are poorly immunogenic in pigs. The characterization of linear sites has contributed to a better understanding of the antigenic structure of SVDV and mAbs used to this purpose may provide a useful tool for the improvement of diagnostic methods, such as antigen detection systems, and analyses of the antigenic profile of SVDV isolates.
Borrego
Belén
B
Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna, Via Bianchi 7, 25125 Brescia, Italy1.
García-Ranea
Juan Antonio
JA
Protein Design Group, Centro Nacional de Biotecnología, Cantoblanco, Madrid, Spain2.
Douglas
Alastair
A
Virology Department, Department of Veterinary Science, Queen's University, Belfast, UK3.
Brocchi
Emiliana
E
Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna, Via Bianchi 7, 25125 Brescia, Italy1.
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D000914:Antibodies, Viral / Q000096:biosynthesis / Q000276:immunology*; D002213:Capsid / Q000737:chemistry / Q000276:immunology*; D003198:Computer Simulation; D029822:Enterovirus B, Human / Q000276:immunology*; D018604:Epitope Mapping; D051379:Mice; D008807:Mice, Inbred BALB C; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000276:immunology; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D011994:Recombinant Proteins; C446935:VP1 protein, Swine vesicular disease virus; D014764:Viral Proteins