IN-VITRO PRODUCTION OF HUMAN ANTISPERM ANTIBODIES - EFFECT OF AN OLIGOCLONAL ANTIBODY (F6) ON SPERM-EGG INTERACTION

A method has been developed to establish lines of transformed lymphocy tes able to produce in vitro the same anti-sperm antibodies as those n aturally occurring in immuno-infertile individuals. We utilized lympho cytes from a male donor whose serum contained anti-sperm antibodies of the IgG class up to the dilution 1:10 000, as detected by means of im munobead binding. T lymphocytes were separated from B lymphocytes usin g magnetic beads coated with anti-T antibody. B lymphocytes were then placed at a concentration of 5 x 10(6)/ml in a 96-well plate, stimulat ed with phytohaemagglutinin (PHA) and transformed with Epstein-Barr vi rus. After a few days, only transformed cells continued growing and th ese were collected. The supernatant was tested for production of anti- sperm antibodies and those transformed lymphocytes shown to be synthes ising antibodies directed against the sperm head and the tail were clo ned. We obtained a clone of cells producing antibodies of the IgG1 cla ss directed against the head of the spermatozoon. This oligoclonal ant ibody (F6) recognized a 58-kDa band from a lysate of sperm membranes a nd was able to reduce the penetration of zona-free hamster oocytes by capacitated spermatozoa.