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

Abstract

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.

Can anti-ovarian antibody testing be useful in an IVF-ET clinic?

“…Frequency of AOA positives in the study groups AOA positive patients' demonstrated immunoreactivity to antigens spanning regions 30-120 kDa in Western blotting as described in details earlier [5]. Of the 570 patients enrolled for this study, 138 were tested AOA positive (24%) by the Western blotting (Fig.…”

“…Although various investigators have demonstrated that there is multiplicity of molecular and cellular targets in human ovarian autoimmunity, there is still a need to identify all specific ovarian targets [29,30]. However, from our earlier findings we have investigated the cause of the non-specificity and were able to demonstrate true molecular and histochemical targets [5,13]. Once the patient tests positive for AOA by Western blot analysis, the serum is screened by us to identity the cell types which are targeted, using IHC which employs a novel blocking recipe [13].…”

“…A total of 570 women (mean age 33 years) presented themselves for infertility treatment at the Department of Assisted Reproduction and Genetics at the Jaslok Hospital and research centre. Inclusion and exclusion criteria for patients enrolled in this on-going study have been described earlier in detail [5]. In brief, women less than 40 years of age at the time of enrollment in the study presenting with infertility and for whom IVF-ET had been determined to be the next treatment for conception.…”

“…The hypothesis of an underlying autoimmune mechanism has been reported in some cases of in vitro fertilization (IVF) failure and in premature ovarian failure (POF) (also known as primary ovarian insufficiency-POI) where the presence of anti-ovarian antibodies (AOA) has been described [1][2][3][4][5][6]. These AOA could affect egg and embryo development and could be responsible for implantation failures.…”

“…This problem of non-specificity has been overcome in a specific, simple and non-invasive diagnostic test developed by us [13]. Using this test we could show new and true molecular and histochemical targets in the ovary in case of ovarian autoimmunity [5,14]. The next step would be to suppress the activity of these autoantibodies before commencing the ovarian stimulation protocol with a standardized dose of corticosteroids.…”