0021-972X/90/7001 -0069$02.00/0 Journal of Clinical Endocrinology and Metabolism Copyright © 1990 by The Endocrine Society
Vol. 70, No. 1 Printed in U.S.A.
Ovarian Antibodies Detected by Immobilized Antigen Immunoassay in Patients with Premature Ovarian Failure* J. L. LUBORSKY, I. VISINTIN, S. BOYERS, T. ASARI, B. CALDWELL, AND A. D E C H E R N E Y
Departments of Obstetrics and Gynecology and Biology (J.L.L.), Yale University School of Medicine, New Haven, Connecticut 06510
gonadotropin antibodies alone do not appear to account for POF. In addition, 2 patients were treated by immunosuppression and became pregnant coincident with a decline in the serum concentration of ovarian antibodies. In summary, the results of this study are consistent with previous immunohistochemical data which indicate that ovarian and oocyte antibodies are common in patients with POF. This supports the concept that some forms of POF are associated with an autoimmune process. Furthermore, detection of ovarian and oocyte antibodies by ELISA may permit routine diagnosis of autoimmune POF and provide a basis for therapy. (J Clin Endocrinol Metab 70: 69, 1990)
ABSTRACT. An enzyme-linked immunosorbent assay (ELISA) was used to detect ovarian and oocyte antibodies in serum from 45 patients with premature ovarian failure (POF). Control sera were obtained from a similar group of normally cycling women without POF. A specific antibody reaction was found when POF sera were tested against human ovary (47%) or oocytes (47%). A combined total of 69% of the sera were positive for either ovary or oocytes. Fewer sera were positive for antibodies against human thyroid (18%) or human placenta (22%), and virtually no reaction with human liver (4%) was seen. LH antibodies were detected by ELISA against LH in only 3 POF sera that also contained ovarian antibodies. Therefore,
P
difficult to assess POF serum against a uniform antigen preparation. For the same reason, comparison between different samples and among samples from the same patient is difficult, and the assessment of multiple POF sera against single tissue sections is laborious. Furthermore, determination of positive staining is a subjective decision of the observer. Finally, it has been suggested that antibodies against gonadotropins could simulate POF (12), but these antibodies would not be detected by immunohistochemical methods. A standardized immunoassay for ovarian antibodies would be useful, but has not been reported previously. The purpose of this study was to develop a routine immunoassay for detection of ovarian and related tissue antibodies.
REMATURE ovarian failure (POF) is generally defined as the loss of ovarian, and therefore reproductive, function before age 40 yr. Several factors, such as irradiation, chemotherapy, and chromosomal anomalies, are known to contribute to this premature loss of function. In addition, there are women who are apparently otherwise healthy and whose ovarian failure is not readily explained. An autoimmune process involving the ovary has been suggested as a basis for POF (1,2) because it is often associated with both endocrine and nonendocrine autoimmune disease (1-4). Furthermore, a lymphocytic infiltrate has been observed in the ovaries of women with POF (1), and ovarian antibodies have been detected in the sera of some women with POF (4-10). The presence of specific tissue autoantibodies is a characteristic feature of autoimmune disease (11). In previous studies of POF, ovarian antibodies were detected by immunohistochemistry. Unfortunately, antigens are not uniformly represented in individual tissue sections, since ovarian follicles in different areas of the ovary are in various functional stages. This makes it
Materials and Methods Human subjects Sera from 45 women with POF were assessed for the presence of ovarian, oocyte, and gonadotropin antibodies. Criteria for POF included persistent high serum gonadotropin levels (LH and FSH, >50 IU/L) or low serum estrogen levels (estradiol,
Vol. 70, No. 1 Printed in U.S.A.
Ovarian Antibodies Detected by Immobilized Antigen Immunoassay in Patients with Premature Ovarian Failure* J. L. LUBORSKY, I. VISINTIN, S. BOYERS, T. ASARI, B. CALDWELL, AND A. D E C H E R N E Y
Departments of Obstetrics and Gynecology and Biology (J.L.L.), Yale University School of Medicine, New Haven, Connecticut 06510
gonadotropin antibodies alone do not appear to account for POF. In addition, 2 patients were treated by immunosuppression and became pregnant coincident with a decline in the serum concentration of ovarian antibodies. In summary, the results of this study are consistent with previous immunohistochemical data which indicate that ovarian and oocyte antibodies are common in patients with POF. This supports the concept that some forms of POF are associated with an autoimmune process. Furthermore, detection of ovarian and oocyte antibodies by ELISA may permit routine diagnosis of autoimmune POF and provide a basis for therapy. (J Clin Endocrinol Metab 70: 69, 1990)
ABSTRACT. An enzyme-linked immunosorbent assay (ELISA) was used to detect ovarian and oocyte antibodies in serum from 45 patients with premature ovarian failure (POF). Control sera were obtained from a similar group of normally cycling women without POF. A specific antibody reaction was found when POF sera were tested against human ovary (47%) or oocytes (47%). A combined total of 69% of the sera were positive for either ovary or oocytes. Fewer sera were positive for antibodies against human thyroid (18%) or human placenta (22%), and virtually no reaction with human liver (4%) was seen. LH antibodies were detected by ELISA against LH in only 3 POF sera that also contained ovarian antibodies. Therefore,
P
difficult to assess POF serum against a uniform antigen preparation. For the same reason, comparison between different samples and among samples from the same patient is difficult, and the assessment of multiple POF sera against single tissue sections is laborious. Furthermore, determination of positive staining is a subjective decision of the observer. Finally, it has been suggested that antibodies against gonadotropins could simulate POF (12), but these antibodies would not be detected by immunohistochemical methods. A standardized immunoassay for ovarian antibodies would be useful, but has not been reported previously. The purpose of this study was to develop a routine immunoassay for detection of ovarian and related tissue antibodies.
REMATURE ovarian failure (POF) is generally defined as the loss of ovarian, and therefore reproductive, function before age 40 yr. Several factors, such as irradiation, chemotherapy, and chromosomal anomalies, are known to contribute to this premature loss of function. In addition, there are women who are apparently otherwise healthy and whose ovarian failure is not readily explained. An autoimmune process involving the ovary has been suggested as a basis for POF (1,2) because it is often associated with both endocrine and nonendocrine autoimmune disease (1-4). Furthermore, a lymphocytic infiltrate has been observed in the ovaries of women with POF (1), and ovarian antibodies have been detected in the sera of some women with POF (4-10). The presence of specific tissue autoantibodies is a characteristic feature of autoimmune disease (11). In previous studies of POF, ovarian antibodies were detected by immunohistochemistry. Unfortunately, antigens are not uniformly represented in individual tissue sections, since ovarian follicles in different areas of the ovary are in various functional stages. This makes it
Materials and Methods Human subjects Sera from 45 women with POF were assessed for the presence of ovarian, oocyte, and gonadotropin antibodies. Criteria for POF included persistent high serum gonadotropin levels (LH and FSH, >50 IU/L) or low serum estrogen levels (estradiol,
