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Biology of Reproduction 61, 1373-1380 (1999)
©Copyright 1999 Society for the Study of Reproduction, Inc.

Articles

Mapping of Dominant B-Cell Epitopes of a Human Zona Pellucida Protein (ZP1)1

Sheri M. Skinnera, Eric S. Schwoebela,b, Sarvamangala V. Prasada, Mary Ogunab, and Bonnie S. Dunbar2,a

a Department of Cell Biology, Baylor College of Medicine, Houston, Texas 77030-3411 b Institute of Primate Research, Nairobi, Kenya

Zona pellucida (ZP) glycoproteins contain numerous antigenic determinants including carbohydrate, protein, and conformational epitopes; and the immunogenicity of these complex glycoproteins varies in different mammalian hosts. Studies have now shown that antibodies from primates immunized with a cDNA-expressed recombinant rabbit ZP protein (the homologue of the human ZP1 [hZP1]) inhibit sperm binding to the ZP without altering ovarian function, unlike immunization with ZP3 and ZP2 protein families. The ZP1 protein or peptides derived from it (recombinant or synthetic) are therefore primary candidates for use in designing safe and reversible human and animal contraceptive vaccines. In order to define peptide epitope(s) that may be critical for eliciting an immune response sufficient to effect immunological contraception without causing any adverse effects on ovarian physiology, studies have been carried out to identify immunodominant B-cell epitopes of the ZP1 protein. The amino acid sequence of the hZP1 was used to design a set of 94 (15-mer) biotinylated peptides having an overlap of 9 amino acids. Using these peptides in a modified enzyme-linked immunoassay, antibodies in sera from rabbits or baboons immunized with native porcine ZP protein were screened for ZP1 peptide recognition.

These studies demonstrate that there are a limited number of peptides recognized by primate antibodies but that the overlapping peptides sharing the sequence GPLTLELQI are recognized by both rabbit and baboon antibodies regardless of the adjuvant system used to induce the immune response. This peptide is 100% conserved in amino acid sequence between the human and pig, although the rabbit protein has two conserved amino acid substitutions (100% similar, 77% identical). Because this peptide is immunogenic as well as antigenic in primates, it could play a major role in the development of human contraceptive vaccines.

1 This project was supported by funding to B.D. from the Contraception and Research Development Program (CONRAD), The Mellon Foundation, and The National Institutes of Health #HD-17543, and by funding to the Institute of Primate Research, Nairobi, Kenya, by the World Health Foundation, The World Bank, and the National Council for Population and Development.

2 Correspondence: Bonnie S. Dunbar, Department of Cell Biology, 112A, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030-3411. FAX: 713 798 7341; bdunbar{at}bcm.tmc.edu






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Copyright © 1999 by the Society for the Study of Reproduction.