Detection of antisperm antibodies: their localization to human sperm antigens that are transferred to the surface of zona-free hamster oocytes during the sperm penetration assay.
The authors have developed an extension of the sperm penetration assay for detecting serum immunoglobulins to sperm antigens that are transferred to the plasma membrane of a sperm-penetrated hamster oocyte. After the hamster oocytes have been scored for sperm penetration by observing for the presence of swollen sperm heads, they are incubated in serum followed by either a 20-minute treatment with rhodamine-conjugated protein A (which binds to most subclasses of IgA, IgG, and IgM) or a 2-hour incubation in guinea pig serum (complement). Positive fluorescence indicates that the serum contains antibodies to sperm antigens that were transferred to the surface of an oocyte during gamete fusion. Complement-mediated lysis indicates that the immunoglobulin that is bound can also fix complement. The advantages of this assay for detection of serum antisperm antibodies are that it is an extension of a widely used assay, is rapid and requires readily available reagents and equipment, can detect most subclasses of IgA, IgG, and IgM, detects antibodies to those sperm antigens that may be transferred to the oocyte during fertilization, and indicates whether the detected antisperm antibodies can mediate complement-dependent lysis of the fertilized oocyte.
D000818:Animals; D000954:Antigens, Surface / Q000032:analysis*; D001323:Autoantibodies / Q000032:analysis*; D003165:Complement System Proteins / Q000276:immunology; D006224:Cricetinae; D003602:Cytotoxicity, Immunologic; D005260:Female; D006801:Humans; D008297:Male; D009865:Oocytes / Q000032:analysis / Q000276:immunology*; D013084:Sperm-Ovum Interactions; D013094:Spermatozoa / Q000276:immunology*
D000954:Antigens, Surface; D001323:Autoantibodies; D003165:Complement System Proteins