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Monoclonal antibodies inducing agglutination of human sperm
17
E. Workshop and Posters I. Antigens of Gametes
MONOCLONAL ANTIBODIES
INDUCING AGGLUTINATION
OF HUMAN SPERM
YUAN CHANG YAN, LIN FANG WANG, EIMEI SATO, AND S.S. KOIDE The Population Council, 1230 York Ave, New York, New York, 10021, U.S.A. Triton X-100 extract of human sperm membrane proteins was prepared. The membrane proteins neutralized the sperm agglutinating activity of serum No. i00 obtained from an infertile woman. (gift of Dr. T. Hjort). Balb/c female mice were immunized with the sperm membrane proteins. Those immunized mice producing sera capable of agglutinating human sperm were selected. The spleen cells of the selected immunized mice were fused with mouse myeloma cells (sp 2/0) by the method of Kohler and Milstein as described by Goding. Several fused cells producing sperm agglutinating antibodies were recloned. One of the fused cells was designated as YWK-I cell line. The YW~-I cells were inoculated into the peritoneal cavity of Balb/c mice and the ascitic fluid collected. Both the conditioned media of YWK-I cell cultures and the ascitic fluid induced tail-to-tail agglutination. The monoclonal antibodies inducing spermagglutination interacted with surface antigens of the midpiece and flagellum of human spermatozoa as determined by an indirect immunofluorescence method. The Triton X-100 sperm extract was subjected to SDS polycrylamide gel electrophoresis and the protein bands transferred to a nitrocellulose sheet. The monoclonal antibodies interacted with a single protein band with an estimated mol. wt. of 84 Kdaltons, identified by an indirect immunofluorescence method. (Supported by grant GAPS 8208 from the Rockefeller Foundation. YCY, LFW, and ES are post-doctoral fellows of the Rockefeller Foundation).
INTERACTION OF GUINEA PIG SPERMATOZOA WITH IMMUNOGLOBULINS : EVIDENCE OF Fc~-SPECIFIC MEMBRANE RECEPTORS ON SPERMATOZOA. Marta De Almeida, Th~r~se Neveu and Bril~itte Marquant-Le Guienne. C.I.P.I.Ex. (U 23 INSERM, LA 289 CNRS, C 16 Ass. CI-Bernard), H6p. St-Antoine, 75571 PARIS Cedex 12 France The ability of guinea pig epididymal spermatozoa to bind homologous immunoglobulins to their surface was investigated by a protein A-coated sheep red blood cells rosetring assay. Under optimal conditions, 47 96 and 32 96 of the spermatozoa were shown to bind respectively the monomeric and the heat-aggregated form ol guinea pig lgG anti-DNP.
E. Workshop and Posters I. Antigens of Gametes
MONOCLONAL ANTIBODIES
INDUCING AGGLUTINATION
OF HUMAN SPERM
YUAN CHANG YAN, LIN FANG WANG, EIMEI SATO, AND S.S. KOIDE The Population Council, 1230 York Ave, New York, New York, 10021, U.S.A. Triton X-100 extract of human sperm membrane proteins was prepared. The membrane proteins neutralized the sperm agglutinating activity of serum No. i00 obtained from an infertile woman. (gift of Dr. T. Hjort). Balb/c female mice were immunized with the sperm membrane proteins. Those immunized mice producing sera capable of agglutinating human sperm were selected. The spleen cells of the selected immunized mice were fused with mouse myeloma cells (sp 2/0) by the method of Kohler and Milstein as described by Goding. Several fused cells producing sperm agglutinating antibodies were recloned. One of the fused cells was designated as YWK-I cell line. The YW~-I cells were inoculated into the peritoneal cavity of Balb/c mice and the ascitic fluid collected. Both the conditioned media of YWK-I cell cultures and the ascitic fluid induced tail-to-tail agglutination. The monoclonal antibodies inducing spermagglutination interacted with surface antigens of the midpiece and flagellum of human spermatozoa as determined by an indirect immunofluorescence method. The Triton X-100 sperm extract was subjected to SDS polycrylamide gel electrophoresis and the protein bands transferred to a nitrocellulose sheet. The monoclonal antibodies interacted with a single protein band with an estimated mol. wt. of 84 Kdaltons, identified by an indirect immunofluorescence method. (Supported by grant GAPS 8208 from the Rockefeller Foundation. YCY, LFW, and ES are post-doctoral fellows of the Rockefeller Foundation).
INTERACTION OF GUINEA PIG SPERMATOZOA WITH IMMUNOGLOBULINS : EVIDENCE OF Fc~-SPECIFIC MEMBRANE RECEPTORS ON SPERMATOZOA. Marta De Almeida, Th~r~se Neveu and Bril~itte Marquant-Le Guienne. C.I.P.I.Ex. (U 23 INSERM, LA 289 CNRS, C 16 Ass. CI-Bernard), H6p. St-Antoine, 75571 PARIS Cedex 12 France The ability of guinea pig epididymal spermatozoa to bind homologous immunoglobulins to their surface was investigated by a protein A-coated sheep red blood cells rosetring assay. Under optimal conditions, 47 96 and 32 96 of the spermatozoa were shown to bind respectively the monomeric and the heat-aggregated form ol guinea pig lgG anti-DNP.