Use of a synthetic peptide adjuvant for the immunization of baboons with denatured and deglycosylated pig zona pellucida glycoproteins.
Baboons were immunized using a synthetic peptide adjuvant with two purified pig zona pellucida glycoproteins. The major zona pellucida glycoprotein (ZPI) was purified by preparative isoelectric focusing, and the 80 K deglycosylated zona pellucida protein (ZPIII) was purified by preparative sodium dodecyl sulfate polyacrylamide gel electrophoresis. The immunogenicity as well as the antigenicity of these proteins were evaluated by characterizing antibodies using the enzyme-linked immunoassay and by immunoblotting of zona pellucida proteins separated by high-resolution two-dimensional polyacrylamide gel electrophoresis. Both groups of animals developed antibodies that recognize the major zona pellucida glycoprotein, (ZPI) and immunoblotting procedures provide evidence that two of the major porcine zona pellucida glycoprotein families (ZPI and ZPIII) contain shared antigenic determinants. The animals immunized with ZPI showed decreased levels of estrogen throughout their menstrual cycles, and two of the animals ceased ovulation. All animals in the group immunized with ZPIII had a significant reduction in the numbers of antral follicles as compared with control animals. Although ovarian cyclicity was not altered significantly within a few months after immunization, two of the five animals in this group became amenorrheic by 8 months. Histologic analysis of ovarian tissue shows that follicles were absent or frequently abnormal in animals of both groups following long-term immunization. These studies demonstrate that the synthetic adjuvant is effective in inducing antibodies (to purified zona pellucida glycoproteins) that recognize antigenic determinants to either denatured or deglycosylated zona pellucida glycoproteins, and that some of these antibodies may interfere with normal ovarian function.
Dunbar
B S
BS
Department of Cell Biology, Baylor College of Medicine, Houston, Texas 77030.
Lo
C
C
Powell
J
J
Stevens
V C
VC
D000276:Adjuvants, Immunologic / Q000276:immunology*; D000818:Animals; D000906:Antibodies / Q000032:analysis; D000941:Antigens / Q000276:immunology; D004527:Egg Proteins; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006023:Glycoproteins / Q000276:immunology* / Q000378:metabolism; D006031:Glycosylation; D007114:Immunization; D015151:Immunoblotting; D008562:Membrane Glycoproteins; D010053:Ovary / Q000033:anatomy & histology / Q000166:cytology / Q000502:physiology; D010215:Papio; D010455:Peptides / Q000276:immunology*; D011956:Receptors, Cell Surface; D013552:Swine; D015044:Zona Pellucida / Q000276:immunology; D000072216:Zona Pellucida Glycoproteins
D000276:Adjuvants, Immunologic; D000906:Antibodies; D000941:Antigens; D004527:Egg Proteins; D006023:Glycoproteins; D008562:Membrane Glycoproteins; D010455:Peptides; D011956:Receptors, Cell Surface; D000072216:Zona Pellucida Glycoproteins