Human antisperm monoclonal antibodies constructed postvasectomy.
Sperm and spermatogenic cell antigens, escaping the blood-testis/blood-epididymal barrier, elicit an autoimmune response in patients following vasectomy. In this study, antisperm antibody-positive sera and peripheral blood lymphocytes were obtained 6-9 mo following vasectomy. Serum antisperm antibody levels were assessed by enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescence. Lymphocyte-myeloma hybridomas were constructed by fusing peripheral blood lymphocytes, harvested from antisperm antibody-positive sera, with a hypoxanthine guanine-phosphoribosyltransferase (HGPRT)-negative mouse myeloma line. Immunoglobulin-secreting colonies surviving drug selection were detected by ELISA and screened for antisperm activity. Antisperm antibody-producing cultures were cloned and expanded for bulk antibody production both in culture and as ascites in athymic nude mice. Eight mouse-human fusions yielded 205 hybridomas secreting human monoclonal antibody, of which 11 demonstrated antisperm reactivity by ELISA. Two of these hybridomas are described in detail: HAS-1, which secretes human immunoglobulin M (IgM, kappa)-recognizing epitopes located on the sperm midpiece, and HAS-2 (IgM, lambda), which secretes monoclonal antibody-recognizing epitopes located on the entire sperm tail. The results indicate successful capture of human antisperm autoantibody from the postvasectomy autoimmune state using somatic cell hybridization techniques.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001323:Autoantibodies / Q000276:immunology*; D005455:Fluorescent Antibody Technique; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D005779:Immunodiffusion; D007621:Karyotyping; D008297:Male; D051379:Mice; D008819:Mice, Nude; D009101:Multiple Myeloma / Q000201:enzymology / Q000473:pathology; D011184:Postoperative Period; D013094:Spermatozoa / Q000276:immunology*; D014659:Vasectomy
D000911:Antibodies, Monoclonal; D001323:Autoantibodies