The proposed research represents a continuation of efforts to critically evaluate an immunocontraceptive approach to fertility regulation and contraceptive vaccine development based upon use of a 55K antigen (ZP3) present in porcine zona pellucida. Biological investigations include examining the longevity of the immune response and antifertility effects produced by active heteroimmmunization of squirrel monkeys with ZP3, determining minimum ZP3 dosage required to elicit zona reactive antibodies, comparing monkey immune response following ZP3 immunization with use of various adjuvants and assessing potential detrimental effects of ZP3 immunization by comparative hormonal (E2, P, LH, FSH) investigations and ovarian histological studies. Squirrel monkey immune response and antibody titers will be monitored by radioimunoassay and in vitro contraceptive potential of antisera assayed by sperm-zona binding assays. Biochemical investigations are directed toward elucidation of the chemical and structural nature of ZP3 antigenic determinants, evaluating the immunogenicity of both deglycosylated derivatives and native and synthetic peptides of ZP3, developing purification protocols for the other porcine zona glycoproteins (ZP1, ZP2, and ZP4) and initiating biochemical and immunological characterization studies on ZP1, ZP2 and ZP4 in anticipation of their future consideration for contraceptive vaccine development ZP3 and other zona components will be isolated using standard chromatographic and HPLC methodology and homogeneity verified by one-and-two dimensional acrylamide gel electrophoresis. Antigenicity and immunoreactivity of isolated components will be determined by radioimmnoassay and electrophoretic blotting. Positive findings from the proposed research would provide critical knowledge for development of a human contraceptive vaccine based upon glycoproteins present in the porcine zona pellucida.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
2R01HD010189-07A1
Application #
3311232
Study Section
Reproductive Endocrinology Study Section (REN)
Project Start
1978-07-01
Project End
1989-06-30
Budget Start
1986-07-01
Budget End
1987-06-30
Support Year
7
Fiscal Year
1986
Total Cost
Indirect Cost
Name
Wayne State University
Department
Type
Schools of Medicine
DUNS #
City
Detroit
State
MI
Country
United States
Zip Code
48202
Yurewicz, E C; Zhang, S; Sacco, A G (1993) Generation and characterization of site-directed antisera against an amino-terminal segment of a 55 kDa sperm adhesive glycoprotein from zona pellucida of pig oocytes. J Reprod Fertil 98:147-52
Yurewicz, E C; Pack, B A; Sacco, A G (1991) Isolation, composition, and biological activity of sugar chains of porcine oocyte zona pellucida 55K glycoproteins. Mol Reprod Dev 30:126-34
Naz, R K; Sacco, A G; Yurewicz, E C (1991) Human spermatozoal FA-1 binds with ZP3 of porcine zona pellucida. J Reprod Immunol 20:43-58
Sacco, A G; Yurewicz, E C; Subramanian, M G et al. (1989) Porcine zona pellucida: association of sperm receptor activity with the alpha-glycoprotein component of the Mr = 55,000 family. Biol Reprod 41:523-32
Sacco, A G; Yurewicz, E C; Subramanian, M G (1989) Effect of varying dosages and adjuvants on antibody response in squirrel monkeys (Saimiri sciureus) immunized with the porcine zona pellucida Mr = 55,000 glycoprotein (ZP3). Am J Reprod Immunol 21:1-8
Yurewicz, E C; Sacco, A G; Subramanian, M G (1987) Structural characterization of the Mr = 55,000 antigen (ZP3) of porcine oocyte zona pellucida. Purification and characterization of alpha- and beta-glycoproteins following digestion of lactosaminoglycan with endo-beta-galactosidase. J Biol Chem 262:564-71
Sacco, A G; Pierce, D L; Subramanian, M G et al. (1987) Ovaries remain functional in squirrel monkeys (Saimiri sciureus) immunized with porcine zona pellucida 55,000 macromolecule. Biol Reprod 36:481-90
Sacco, A G; Yurewicz, E C; Subramanian, M G (1986) Carbohydrate influences the immunogenic and antigenic characteristics of the ZP3 macromolecule (Mr 55 000) of the pig zona pellucida. J Reprod Fertil 76:575-85