title: Antiacrosin antibodies and infertility. II. Gene immunization with human proacrosin to assess the effect of immunity toward proacrosin/acrosin upon protein activities and animal fertility.
abstract: OBJECTIVE
  To assess the effect of antiacrosin antibodies upon proacrosin/acrosin activities and animal fertility.
  
  
  METHODS
  Prospective study.
  
  
  METHODS
  Basic research laboratory.
  
  
  METHODS
  A gene immunization (GI) model was developed; mice were injected with the sequence encoding human proacrosin (h-proacrosin), cloned in an expression vector.
  
  
  METHODS
  Subcloning of h-proacrosin in a eukaryotic expression vector (promoter, CMV; leader sequence, alpha-1 antitrypsin; pSF2-Acro); GI of female mice with this plasmid.
  
  
  METHODS
  The following parameters were evaluated: [1] adequate conditions for GI protocols, [2] humoral response to GI with pSF2-Acro, [3] protein regions recognized by the antibodies, and [4] effect of antibodies upon proacrosin/acrosin-ZPA binding and amidase activity, and animal fertility.
  
  
  RESULTS
  Conditions of female mice GI with the proacrosin sequence were established (plasmid purification with anion exchange chromatography and 40 microg of pSF2-Acro per dose) to trigger an immune response, reaching maximum levels at week 9 after the first injection. Antibodies produced by GI recognized human and mouse sperm acrosin systems, inhibited human proacrosin/acrosin interaction with recombinant human ZPA and protease activity, and negatively affected mouse IVF and early embryonic development. In addition, mice immunized with SF2-Acro exhibited a significantly lower size of fetuses.
  
  
  CONCLUSIONS
  Antiacrosin antibodies developed by using GI inhibit human proacrosin/acrosin activities and impair mouse fertility.
authors.affiliation: Instituto de Biología y Medicina Experimental, Consejo Nacional de Investigaciones Científicas y Técnicas-Universidad de Buenos Aires, Buenos Aires, Argentina.
authors.forename: Carolina
authors.initials: C
authors.lastname: Veaute
authors.forename: Laura I
authors.initials: LI
authors.lastname: Furlong
authors.forename: Mónica
authors.initials: M
authors.lastname: Cameo
authors.forename: Jeffrey D
authors.initials: JD
authors.lastname: Harris
authors.forename: Mónica H
authors.initials: MH
authors.lastname: Vazquez-Levin
mesh_terms: D000176:Acrosin / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D000818:Animals; D001323:Autoantibodies / Q000097:blood / Q000494:pharmacology / Q000502:physiology*; D047108:Embryonic Development / Q000187:drug effects / Q000276:immunology; D004792:Enzyme Precursors / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D005260:Female; D005298:Fertility / Q000235:genetics / Q000276:immunology*; D005306:Fertilization / Q000235:genetics / Q000276:immunology; D006801:Humans; D007114:Immunization / Q000009:adverse effects; D007246:Infertility / Q000097:blood / Q000209:etiology* / Q000276:immunology / Q000662:veterinary; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D011247:Pregnancy
chemical_list: D001323:Autoantibodies; D004792:Enzyme Precursors; C022438:proacrosin; D000176:Acrosin