ABSTRACT
Porcine parvovirus (PPV) is a DNA virus that causes reproductive failure in gilts and sows, resulting in embryonic and fetal losses worldwide. Epitope mapping of PPV is important for developing new vaccines. In this study, we used spot synthesis analysis for epitope mapping of the capsid proteins of PPV (NADL-2 strain) and correlated the findings with predictive data from immunoinformatics. The virus was exposed to three conditions prior to inoculation in pigs: native (untreated), high hydrostatic pressure (350 MPa for 1 h) at room temperature and high hydrostatic pressure (350 MPa for 1h) at −18 °C, compared with a commercial vaccine produced using inactivated PPV. The screening of serum samples detected 44 positive spots corresponding to 20 antigenic sites. Each type of inoculated antigen elicited a distinct epitope set. In silico prediction located linear and discontinuous epitopes in B cells that coincided with several epitopes detected in spot synthesis of sera from pigs that received different preparations of inoculum. The approach used here provided important information on the antibody/antigen interactions required to improve B cell responses to PPV and may be useful in developing novel strategies for producing new vaccines.
Abbreviations 3D, three dimensional; HHP, high hydrostatic pressure; ID, identification; IEDB, Immune Epitope Database; ORFs, open reading frames; p.i., post-infection; PPV, porcine parvovirus; RMSD, root-mean-square deviation of atomic positions; R(g), radius of gyration. RMSF, root-mean-square fluctuation; SK6, Swine kidney cell; SPF, specific pathogen free; TCID50/mL, Median Tissue Culture Infectious Dose; HI, Hemagglutination Inhibition.
