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Monoclonal antibody definition of sperm — specific antigen crossreactivity between sea urchin and mammals
21 bound less C3 than did hamster caput spermatozoa. Reduction during epididymal maturation in the tendency of spermatozoa to activate complement may bear on the response of the female tract in a way that affects transport and survival of spermatozoa there, following insemination.
IMMUNOLOGICAL SPECIFICITY OF ~-N-ACETYL GLUCOSAMINIDASES FROMHUMAN SEMINAL PLASMA. Gopal S. Gupta and Deepak K. Kaput Department of Biophysics, Panjab University, Chandigarh, India. ~-N-acetyl glucosaminidase (~-N-Ac Glndase) of sperm acrosome is considered as one of the important enzymes for sperm capacitation and fertilization of ovum. Though ~-N-Ac Glndase in seminal plasma (SP) appears to originate from sperm aerosome, there is no report regarding its immunological specificity and immunoehemical characterization. In view of this, two isozymes of ~-N-Ac Glndase were purified from human SP. Antibodies produced in rabbits of each isozyme showed homogenity of each isozyme by immunoelectrophoresis and immunodiffusion techniques. Anti-serum of isozyme A cross-reacted with isozyme B and vice-versa. Thus common antigenic determinants were indicated in two isozymes of ~-N-Ac Glndase. Moreover, antibodies of both forms of ~-N-Ac Glndase cross-reacted with human sperm, testis, epididymis, prostate, spleen, kidney, lung, liver, pancreas, gallbladder and intestine, suggesting that there is nothing like semen specific enzyme of ~-N-Ac Glndase. Though monkey tissues also cross-reacted with human ~-N-Ac Glndase antisera, mouse, rat and goat testis and epididymis extracts failed to show any crossreaction. These studies reveal that human SP ~-N-Ac Glndase does not contain tissue specific antigenic determinants which may be linked to its specific function. It is likely that the specific function of sperm ~-N-Ac Glndase is related to one of the subunit structures of one or both isozymes. Immunochemical properties of two isozymes from human SP are under progress. MONOCLONAL ANTIBODY DEFINITION OF SPERM - SPECIFIC ANTIGEN CROSSREACTIVITY BETWEEN SEA URCHIN AND M ~ L S Padma Shastry, Bagirath Singh, Marilyn Hamilton and Thomas G.Wegmann Dept. of Immunol., Univ. of Alberta, Edomonton, Canada Dept. of Ultrastructure, Univ. of Washington, Seatle, Washington, U.S.A. LOPO and VACQUIER reported that rabbits immunized with sperm from sea urchins (Strongylocentrotus Purpuratus) made antibody that reacted with sperm from 28 different species, representing seven different phyla (Nature 288 : 397, 1980). Using a solid phase assay, we have found that Balb/CCR female mice make a very strong anti-sea urchin sperm antibody response after only two immunizations. The antiserum crossreacts with human, mouse, rat and boar sperm. We therefore proceeded to make hybridomas, and to date have obtained nine clones which stably produce crossreactive monoclonal antibody against sperm. Five of these monoclonal antibodies have been tested for crossreactivity with tissues other than sperm, by absorption, followed by solid phase assay on mouse epididymal sperm. They did not react with brain, thymus, spleen, liver, testis, bone marrow, or erythrocytes, but they did absorb out with mouse epididymal sperm. One monoclonal antibody was absorbed with mouse epididymal sperm and then tested against human, boar and sea urchin sperm. A decreased titer resulted with all three antigens. These data indicate that there is a very immunogenic antigen present on sea urchin sperm that has been apparently highly conserved in evolution, and that may be sperm specific.
IMMUNOLOGICAL SPECIFICITY OF ~-N-ACETYL GLUCOSAMINIDASES FROMHUMAN SEMINAL PLASMA. Gopal S. Gupta and Deepak K. Kaput Department of Biophysics, Panjab University, Chandigarh, India. ~-N-acetyl glucosaminidase (~-N-Ac Glndase) of sperm acrosome is considered as one of the important enzymes for sperm capacitation and fertilization of ovum. Though ~-N-Ac Glndase in seminal plasma (SP) appears to originate from sperm aerosome, there is no report regarding its immunological specificity and immunoehemical characterization. In view of this, two isozymes of ~-N-Ac Glndase were purified from human SP. Antibodies produced in rabbits of each isozyme showed homogenity of each isozyme by immunoelectrophoresis and immunodiffusion techniques. Anti-serum of isozyme A cross-reacted with isozyme B and vice-versa. Thus common antigenic determinants were indicated in two isozymes of ~-N-Ac Glndase. Moreover, antibodies of both forms of ~-N-Ac Glndase cross-reacted with human sperm, testis, epididymis, prostate, spleen, kidney, lung, liver, pancreas, gallbladder and intestine, suggesting that there is nothing like semen specific enzyme of ~-N-Ac Glndase. Though monkey tissues also cross-reacted with human ~-N-Ac Glndase antisera, mouse, rat and goat testis and epididymis extracts failed to show any crossreaction. These studies reveal that human SP ~-N-Ac Glndase does not contain tissue specific antigenic determinants which may be linked to its specific function. It is likely that the specific function of sperm ~-N-Ac Glndase is related to one of the subunit structures of one or both isozymes. Immunochemical properties of two isozymes from human SP are under progress. MONOCLONAL ANTIBODY DEFINITION OF SPERM - SPECIFIC ANTIGEN CROSSREACTIVITY BETWEEN SEA URCHIN AND M ~ L S Padma Shastry, Bagirath Singh, Marilyn Hamilton and Thomas G.Wegmann Dept. of Immunol., Univ. of Alberta, Edomonton, Canada Dept. of Ultrastructure, Univ. of Washington, Seatle, Washington, U.S.A. LOPO and VACQUIER reported that rabbits immunized with sperm from sea urchins (Strongylocentrotus Purpuratus) made antibody that reacted with sperm from 28 different species, representing seven different phyla (Nature 288 : 397, 1980). Using a solid phase assay, we have found that Balb/CCR female mice make a very strong anti-sea urchin sperm antibody response after only two immunizations. The antiserum crossreacts with human, mouse, rat and boar sperm. We therefore proceeded to make hybridomas, and to date have obtained nine clones which stably produce crossreactive monoclonal antibody against sperm. Five of these monoclonal antibodies have been tested for crossreactivity with tissues other than sperm, by absorption, followed by solid phase assay on mouse epididymal sperm. They did not react with brain, thymus, spleen, liver, testis, bone marrow, or erythrocytes, but they did absorb out with mouse epididymal sperm. One monoclonal antibody was absorbed with mouse epididymal sperm and then tested against human, boar and sea urchin sperm. A decreased titer resulted with all three antigens. These data indicate that there is a very immunogenic antigen present on sea urchin sperm that has been apparently highly conserved in evolution, and that may be sperm specific.