Topographical analysis of canine parvovirus virions and recombinant VP2 capsids.
The distribution of epitopes defined by monoclonal antibodies (MAbs) on the surface of canine parvovirus (CPV) virions and recombinant VP2-capsids was established using immunoelectron microscopy. A correlation appeared to exist between the linear position, neutralizing activity and immunogold staining. Both viral capsids and recombinant capsids gave similar patterns of immunostaining. The neutralizing MAbs that recognized epitopes not previously identified by Pepscan or immunoblotting gave a clear staining. However, MAbs 3C9 and 3C10, identified by Pepscan and immunoblotting as recognizing linear epitopes, did not show any labelling (3C9) or only scattered labelling (3C10). MAb 3C9 recognizes an N-terminal domain of VP2. MAb 4AG6, which recognizes the same linear epitope as 3C10, did not bind to the capsids, indicating a different orientation. An immunofluorescence assay was performed to supplement the B cell epitope characterization. In contrast to other MAbs that gave nuclear and cytoplasmic staining, MAb 3C9 gave a preferential nuclear staining. Based on these results, it is hypothesized that the N terminus of VP2 is barely, or not at all, exposed on the surface of the native virions, but becomes accessible after some virion steric change (e.g. after attachment to the cell receptor).
Cortes
E
E
Centro de Biologia Molecular, CSIC-UAM, Cantoblanco, Madrid, Spain.
San Martin
C
C
Langeveld
J
J
Meloen
R
R
Dalsgaard
K
K
Vela
C
C
Casal
I
I
D000818:Animals; D000911:Antibodies, Monoclonal; D000956:Antigens, Viral / Q000032:analysis*; D002213:Capsid / Q000032:analysis* / Q000276:immunology; D036022:Capsid Proteins; D002478:Cells, Cultured; D004285:Dogs; D000939:Epitopes / Q000032:analysis; D005455:Fluorescent Antibody Technique; D015151:Immunoblotting; D016253:Microscopy, Immunoelectron; D009500:Neutralization Tests; D010321:Parvoviridae / Q000648:ultrastructure*; D011994:Recombinant Proteins / Q000032:analysis* / Q000276:immunology; D013347:Subcellular Fractions / Q000648:ultrastructure; D014771:Virion / Q000648:ultrastructure*
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D036022:Capsid Proteins; D000939:Epitopes; D011994:Recombinant Proteins