Ovarian Failure andAutoimmunity Detection ofAutoantibodies Directed against BoththeUnoccupied Luteinizing Hormone/Human Chononic Gonadotropin Receptor andtheHormone-receptor ComplexofBovineCorpusLuteum

We developed anELISAsystem forthedetection ofhuman anti-ovarian antibodies. Bovine corpora lutea wereextracted in PBS(pH7.2) andfractionated byultracentrifugat ion. Boththe soluble fraction obtained after 80,000 g(S80) andtheTritonextracted membrane fraction (ST288) wereusedasantigens. Additionally, theluteinizing hormone (LH)/human chorionic gonadotropin (hCG) receptor wasisolated byaffinity chromatography (wheat germagglutinin andLH-Sepharose) andalso usedasanantigen. In7of14patients withprimary sterility andendometriosis apositive reaction wasobserved. Similarly, 6of16patients withsecondary sterility andendometriosis werealso positive. Patients being stimulated forinvitro fertilization andpresentingeither primary orsecondary sterility werepositive in5of 22and6of16cases, respectively. IntheS80test 41of60sera presented IgG2antibodies, whereas intheST288test 38of60 belonged totheIgG, subclass. Kappaandlambda chains were equally distributed. Somepatients couldrecognize theunoccupied LH/hCG receptor asanantigen, while others recognized only thecomplex formed bythehormone plus thehormone receptor. TheS80andST288antigens wereisolated byaffinity chromatography. Gelpermeation ofthepurified antigens revealed ineachcase thepresence ofanantigen complex. Theapparent molecular weight wasbetween 2,000 and36,000 D.Cross-reactivity studies using affinity-purifie d antibodies demonstrated anantigenic relationship ofthemembrane, soluble, andextractable fractions. NAc-(beta-1 - 4)-D-glucosaminide and -D-galactopyranoside werethemainterminal glycosides.