Studies of a sperm/placenta cross-reacting antigen, STX-10

Abstract A monoclonal antibody, HSA-10 initially produced against acrosome-reacted human sperm was also shown to cross-react with human placenta/trophoblast. Transmission electron microscopy, as well as indirect immunofluorescent assay, demonstrated that HSA-10 was found to react with antigen on the inner acrosome of human sperm. The cognate antigen, designated as STX-10, was found to exist as an aggregate in the native form when analyzed by Sephacryl S-300 gel filtration chromatography. When purified by HSA-10-immunoaffinity chromatography from human placenta extract, STX-10 was found to be predominantly a group of glycoproteins with a subunit molecular mass in the range of 75 ± 5 kDa , whereas an additional group of three proteins with subunit molecular mass less than 20 kDa were co-purified from human sperm extract. A sandwich enzyme immunoassay was designed to quantitatively determine the immunoactivity of STX-10 in solution, using HSA-10 monoclonal antibody for coating and for signal detection via enzyme conjugation. Based on this assay, it was found that STX-10 could be detected only in human sperm and placenta extract, but not in any other human somatic tissues, such as serum, brain, heart, muscle, kidney and liver. The immunoactivity of STX-10 was found to be sensitive to proteolytic digestion, low pH, in the presence of reducing agent, but resistant to treatment with sodium periodate. This observation suggests that HSA-10 specific epitope is a peptide in nature and not a carbohydrate moiety. Results of antifertility studies revealed that HSA-10 significantly inhibited human sperm penetration to zona-free hamster ova. Thus, the results of this study are consistent with those of WHO Workshop evaluation that seem to suggest that STX-10 is a highly gametespecific antigen localized on the inner acrosome of human sperm and in human trophoblast/placenta. Therefore, it may play an important role during human fertilization and embryo development.