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Specific and Sensitive Immunoassays Detect Multiple Anti-ovarian Antibodies in Women With Infertility

Eusebio S. Pires, Pervin K. Meherji, Rama R. Vaidya, Firuza R. Parikh, Manish N. Ghosalkar and Vrinda V. Khole

Department of Gamete Immunobiology (ESP,MNG,VVK) and Reproductive Endocrinology and Infertility Clinic (PKM), National Institute for Research in Reproductive Health, Mumbai, India; Bhartiya Vidya Bhavan Swami Prakashananda Ayurvedic Research Centre, Mumbai, India (RRV); and Department of Assisted Reproduction and Genetics, Jaslok Hospital and Research Centre, In Vitro Fertilization Clinic, Mumbai, India (FRP)

Correspondence to: Dr. Vrinda V. Khole, PhD, Deputy Director (Sr Grade) and Divisional Head, Department of Gamete Immunobiology, National Institute for Research in Reproductive Health, J M Street, Parel, Mumbai 400 012, India. E-mail: kholevv{at}icmr.org.in

Serum anti-ovarian antibodies (AOAs) have been shown in autoimmune premature ovarian failure and in vitro fertilization–embryo transfer (IVF-ET) cases. The specificity of assays detecting these antibodies has been questioned. Researchers have used several techniques (e.g., ELISA and indirect immunofluorescence). Few have reported on the non-specificity and the type of molecular and cellular targets. We reported earlier on the presence of naturally occurring anti-albumin antibodies as the likely factor for non-specificity. Having developed a novel blocking recipe, we show substantial elimination of this non-specificity. With these standardized tests, we hereby report multiple targets at protein and histological levels. In our study group, 15 of 50 (30%) patients with premature ovarian failure and 13 of 50 (26%) IVF-ET patients showed the presence of AOAs. Western blotting showed a large number of patients making AOAs to a 90-kDa protein, followed by 97- and 120-kDa proteins. Histochemically, it was evident that the sera of these patients predominantly react with the oocyte; other somatic cellular targets are also involved. The specific non-invasive test developed by us was found to be useful because it could carry out a reliable diagnosis of an autoimmune etiology that would be very helpful to select patients in whom immune-modulating therapy could be recommended, which in turn may restore ovarian function and fertility. (J Histochem Cytochem 55:1181–1190, 2007)

Key Words: anti-ovarian antibodies • clinical significance • cellular targets • premature ovarian failure • in vitro fertilization–embryo transfer

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