title: Antifertility effect of passive administration of antibodies to 80 kDa human sperm antigen and its synthetic peptides in male and female rats.
abstract: OBJECTIVE
  A human sperm antigen of molecular size of about 80 kDa (80 kDa HSA) has been reported to be sperm-specific, conserved and responsible for inducing immunological infertility. The partial N-terminal amino acid sequence of 80 kDa HSA (peptide NT) and its peptides obtained by enzymatic digestion with endoproteinase Lys-C (peptides 1-4) and with endoproteinase Glu-C (peptides 5 and 6) did not show sequence homology with any of the proteins of the GenBank. The peptides NT, 1, 2, 3 and 4 were synthesized, conjugated to keyhole limpet hemocyanin and used as an immunogen to raise the antibodies in rabbits. Peptide 3 did not elicit significant antibody titer while peptides NT, 1, 2 and 4 elicited significant antibody titer and immunobiologically mimicked the native protein.
  
  
  METHODS
  Effects of passive administration of two injections each of 200 microL of antibodies or 10 and 40 microg purified immunoglobulins to 80 kDa HSA, peptides NT, 1, 2 and 4 on fertility in male and female rats were investigated.
  
  
  RESULTS
  Passive administration of antibodies to 80 kDa HSA and its peptides NT, 1, 2 and 4 resulted in agglutination of epididymal spermatozoa with loss of motility but had no effect on sperm count or weights of the reproductive organs. These animals failed to impregnate normal female rats. Passive administration of these antibodies to female rats also resulted in infertility. The presence of antibodies was detected by enzyme-linked immunosorbent assay in uterine secretions of animals treated with antipeptide antibody. The presence of agglutinated spermatozoa was observed in the post-coital vaginal smears of these animals. The immunized females were found to be ovulating normally and the number of corpora lutea were unaltered. Of the four antipeptide antibodies studied, antibodies to peptides NT and 1 were most effective in inhibiting fertility both in male as well as female rats. Hence, the antifertility studies were further confirmed by passive administration of 10 and 40 microg of purified immunoglobulins of antibodies to NT and 1, which resulted in a dose-dependent inhibition of fertility in male and female rats.
  
  
  CONCLUSIONS
  The study demonstrated that the synthetic peptides of 80 kDa HSA immunobiologically mimicked the native protein and impaired fertility following passive administration of antipeptide antibodies and hence, suggested the suitability of synthetic peptides of 80 kDa HSA as candidates for development of antifertility vaccine.
authors.affiliation: National Institute for Research in Reproductive Health, Parel, Mumbai, India. batmaram@hotmail.com
authors.forename: Atmaram H
authors.initials: AH
authors.lastname: Bandivdekar
authors.forename: Vandana J
authors.initials: VJ
authors.lastname: Vernekar
authors.forename: Masaharu
authors.initials: M
authors.lastname: Kamada
authors.forename: Vijaya P
authors.initials: VP
authors.lastname: Raghavan
mesh_terms: D000372:Agglutination Tests; D000818:Animals; D000906:Antibodies / Q000008:administration & dosage* / Q000737:chemistry / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology; D000937:Antigen-Antibody Reactions / Q000276:immunology; D000941:Antigens / Q000737:chemistry / Q000276:immunology*; D000954:Antigens, Surface / Q000737:chemistry / Q000276:immunology*; D004306:Dose-Response Relationship, Immunologic; D005260:Female; D005298:Fertility / Q000187:drug effects* / Q000276:immunology; D006801:Humans; D007114:Immunization; D008297:Male; D008970:Molecular Weight; D009929:Organ Size / Q000187:drug effects / Q000276:immunology; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology*; D011817:Rabbits; D051381:Rats; D013076:Sperm Count; D013094:Spermatozoa / Q000276:immunology*; D013737:Testis / Q000187:drug effects; D014599:Uterus / Q000187:drug effects
chemical_list: D000906:Antibodies; D000941:Antigens; D000954:Antigens, Surface; D010455:Peptides; C509458:sperm antigen 80-kDa, human