Objective:Use of synthetic zona peptides as target immunogens is a promising approach to an anti-fertility vaccine. To elucidate contraceptive epitopes, we raised polyclonal antibodies against synthetic peptides of pig ZP3α and evaluated for immunogenicity, ability to inhibit in vitro boar sperm-pig zona attachment, and cross-reactivity with human zonae.
Methods:Five synthetic peptides were chosen based on hydropathicity analysis of ZP3α, synthesized and coupled to keyhold limpet haemocyanin (KLH). Pairs of male rabbits were immunized with each peotide-KLH conjugate using a multi-site intradermal injection protocol. Resulting antisera were evaluated for immunoreactivity with cognate peptide and ZP3α/EBGD (purified endo-beta-galactosidase-digested ZP3α), contraceptive potential using an in vitro pig gamete bioassay, and cross-reactivity with human zonae using indirect immunofluorescence.
Results:All anti-peptide-KLH sera demonstrated immunoreactivity with cognate peptide. Anti-peptide sera, except for anti-α-3-KLH serum, were less immunoreactive with ZP3α/EBGD. Antisera directed against α-1-KLH, α-2-KLH, and α-3-KLH demonstrated significant (P < .00001) inhibition of boar sperm-pig zona attachment. Only anti-α-2-KLH and anti-α-3-KLH sera cross-reacted with human zonae.
Conclusion:Synthetic peptides of pig ZP3α are immunogenic, elicit antibodies cross-reactive with human zonae, and have in vitro contraceptive activity. These data support continued investigation of synthetic ZP3α peptides as potential target immunogens for anti-fertility vaccine development.
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