Antiacrosin antibodies. II. Genetic immunization with human proacrosin to assess the effect of immunity towards proacrosin/acrosin upon protein activities and mouse fertility

VEAUTE C; FURLONG LI; CAMEO M; HARRIS JD; VAZQUEZ-LEVIN MH

FERTILITY AND STERILITY

Elsevier

Lugar: Nueva York, NY, EEUU; Año: 2009 vol. 91 p. 1256 - 1268

0015-0282

Study objective: To assess the effect of antiacrosin antibodies upon proacrosin/acrosin activities and animal fertility. Design: Prospective study. Setting: Basic research laboratory. Patients: A gene immunization (GI) model was developed injecting mice with the sequence encoding human proacrosin (h-proacrosin) cloned in an eukaryotic expression vector. Interventions: Subcloning of h-proacrosin in a eukaryotic expression vector (promoter: CMV; leader sequence: alpha–antitrypsin I; pSF2-Acro), and GI of female mice with this plasmid. Main outcome measure(s): Evaluation of the: 1) Humoral response to GI with pSF2-Acro, 2) Adequate conditions for GI protocols, 3) Protein regions recognized by the GI antibodies, and 4-5) Effect of antiacrosin antibodies upon proacrosin/acrosin-ZPA binding/amidase activity and animal fertility. Results: GI of female mice with the proacrosin sequence triggered an immune response, reaching maximum levels on the week 9 after the first injection. Conditions of immunization were established using anion exchange chromatography for plasmid purification and 40 ug of plasmid/dose. Antibodies produced by GI 1) recognized human/mouse sperm protease systems, 2) inhibited proacrosin/acrosin interaction with rec-hZPA and protease activity 3) negatively affected mouse in vitro fertilization (IVF) and early embryonic development. In addition, 4) immunized mice exhibited a decreased number and size of fetuses. Conclusions: Antiacrosin antibodies developed using GI inhibit the human proacrosin/acrosin activities and impair mouse IVF and early embryonic development.