ESTABLISHMENT OF HYBRIDOMA CELL LINES (G8,E10) SECRETING MONOCLONAL ANTIBODIES AGAINST HUMAN SPERMATOZOA / 中国法医学杂志

ABSTRACT

Two hybridoma cell lines (G8,E10) secreting monoclonal antibodies against human spermatozoa were established by fusing Sp2/0 mouse myeloma cells with splenocytes from Balb/c mice immunized with washed human spermatozoa. The results of the IIF, Elisa or peroxidase conjugated antibody im- munohistochemical staining demonstrated that both antibodies reacted with the spermatozoa in the human semen, testis, epididymus and vas deferens as well as the germ cells in the testis, cross reacted with the epithelial cells Of the thyroid gland, renal tubules, pancreatic ducts as well as the ilets of Langhans of the pancrease and not cross reacted with 33 normal human tissue cells, 9 different sources of body fluids and secretions as well as 8 different species of animal's spermatozoa. It is concluded that both monoclonal antibodies can be used for the species identification of semen stain in sex assault cases. The results of IIF demonstrated that each antibody bind to the localized region of the sperm cell surface. The antibody binding patterns were restricted to the acromosal cap, equatorial segment, postacromosal region, neck and the midpiece. The observed antibody binding patterns suggest that the human sperm surface is divided into a minimum of five domains. Both monoclonal antibodies were belong to IgG_1 subclass. The titre of Gs was 1∶512, while of E_(10) 1∶1024.