|Year : 1999 | Volume
| Issue : 6 | Page : 254-258
Antiovarian antibody in premature ovarian failure
D Chattopadhyay1, MR Sen2, P Katiyar3, LK Pandey4
1 Department of Immunology, Institute of Medical Sciences, Banares Hindu University, Varanasi - 221 005., India
2 Department of Microbiology, Institute of Medical Sciences, Banares Hindu University, Varanasi - 221 005., India
3 Department of Gynaecology, , Institute of Medical Sciences, Banares Hindu University, Varanasi - 221 005., India
4 Department of obstetrics and Gynaccology, Institute of Medical Sciences, Banares Hindu University, Varanasi - 221 005., India
Department of Immunology, Institute of Medical Sciences, Banares Hindu University, Varanasi - 221 005.
|How to cite this article:|
Chattopadhyay D, Sen M R, Katiyar P, Pandey L K. Antiovarian antibody in premature ovarian failure. Indian J Med Sci 1999;53:254-8
| ¤ Introduction|| |
The ovary was first documented as target of autoimmunity over three decades ago, yet today the aetiology and pathogenesis of autoimmune mediated premature ovarian failure (POF) are poorly understood. Autoimmune factors may play an etiological role in patients. , POF is associated with a variety of autoimmune disorders such as Addison's disease,  thyroid disorder. , Works of these authors have shown antiovarian autoantiboies presence in women with premature ovarian failure. ,, Ovarian failure occurs in high proportion of women with autoimmune diseases  The demonstration of circulating antibodies to ovarian tissue of women with premature ovarian failure was explained.  The incidence of antiovarian antibodies has varied in different studies according to the types of patient studied and the technique used to detect the antiovarian antibody. Indirect immunofluorescence technique was used on human ovarian tissue. The incidence of the antiovarian antibodies was 35 to 51% in patients with Addison's disease who showed positive antiadrenal antibody.  These findings suggests that, antiovarian antibody may play a role in or refect a n autoimmune process responsible for the development of ovarian failure only in the presence of a profound or generalized altered immunologic reactivity to homologous tissue antigens. The presence of antiovarian antibodies in sera of women with premature ovarian failure by an indirect fluorescent antibody assay using human ovarian tissue is seen.  24 patients positive for non-organ specific and organ specific autoantibodies are studied in order to establish a relationship between POF and autosensitization.  In our present study the presence of antiovarian antibody in the test sera was detected by immunoperoxidase technique  with slight modification.
| ¤ Material and Method|| |
Test sera, ovarian tissue cryostat sections, antimouse gammaglobulin, horseradish peroxidase, 1% glutaldehyde, 3-3' DAB, 3%, H2 O2 , 0.05 mol/L trio HCI buffer, dialysis membrane bag, graded alcohol, xylene haernatoxylene, DPX and coverglasses. Preparation of conjugated gammaglobulin by one step glutaldehyde method. Five of enzyme was dissolved in 1 ml of antibody (2.5 mg/ml) solution dialysed against 0.1 M PBS pH 7.2). 50 ml of 1% gluttraldehyde was added to the solution with gentle sterring and kept for 2-3 hrs at room temp. Solution was dialysed against PBS overnight at 40C and centrifuged for 30 mins at 10,000 rpm at 4oC. Supernatent (conjugate) was stored at 4oC.
Preparation of DAB Solution
(DAB) Diaminobenzedine tetrahydrochloride = 25 mg., tris - HCl/ Buffer = 50 ml., 3%, H2O2 = 0.15 ml, DAB was dissolved in buffer and H2O2 was added immediately fore staining. Immunoperoxidase Staining : Cryostat sections were prewashed for one hour in PBS (pH 7.2). 50 ml of test serum was added to cover cryostat sections and incubated at room temperature in a moist chamber for 30 mins. The sections were washed twice for 5 rains in PBS. Excess PBS was wiped off and 50 ml of conjugate diluted 1 : 20 was added to the sections which then incubated at room temperature in moist chamber of 30 mins. The sections were washed three timse in PBS. The slides were transfered into the substrate solution (freshly prepared) and left for 5 mins. Slides were then washed for 5 mins in distilled water. One batch of slides were tilted with 70%, 90%, 100% alcohol with 2 washes of xylene and mounted under coverglass using u PX. Other batch of slides was stained with conterstain haematoxylene for 1 min. Slidse were washed for 5 mins in distilled water and then treated for about 1 min with each of 70%, 90% 100% alcohol and 2 washes of xylene and mounted over converglass with DPX. Slides were studied under low power and high power objectives for presence of any dark brown pigment sections with dark brown pigment deposition at different antigenic sites were taken as positive.
| ¤ Result|| |
In the present study total 38 patients including 18 patients with POF (study group), 10 regularly menstruating (control group I) and 10 post menopausal women (control group II) were studied. Study group covers 47.4% while control group I and II covers 26.3%. in the study group (16.67%) patients showed antiovarian antibodies while in controls group none were positive [Table 1] shows relation of LH and FSH in presence of antovarian antibodies in study group.
| ¤ Discussion|| |
Premature ovarian failure (POF), is a syndrome consisting of hypergonadotropic amenorrhoea and hypoestrogenism in women under the age of 40 years. There is a close association of autoimmune disorders with POF. ,[ 7] Evidence of autoimmunity is present in 12-18% of patients with POF. , The incidence of presence of antiovarian antibody of POF patients has been reported by many authors in previous studies. Immune disturbances were noted in upto 35% of women with POF. Extraovarian endocrine (specially adrenocortical) involvement is frequent in patients with premature ovarian failure, 49°x, patients had extraovarian endocrine abnormalities, 22% had abnormal thyroid function tests and 41% patients tested for adrenocortical regime showed impared response of plasma cortisol to adrenocorticotropic hormone stimulation. None of the patients had overall clinical evidence of thyroid or adrenocortical disease. The range of antiovarian antibody in sera of POP patients varied between 2.2% to 51.8%. In present study we have studied 18 cases of POF without any overt manifestation of autoimmune disorder. The antiovarian antibody was detected with the idea that this autoantibody might be the cause of ovarian dysfunction which is evident in POF. Sera from 18 patients with 10 regularly menstrutaing women and 10 post menopausal women were examined for presence of antiovarian antibody by immunoperoxidase technique using cryostat section from ovary as substrate. Only 3 cases out of 18 (16.67%) showed positive result for antiovarian antibody in test sera, rest 15 cases were negative for antiovarian antibody. Both the control group cases did not show a single positive result. In our study the percentage of positivity is on the lower side of the range which may be firstly due to non-inclusion of any known autoimmune disorder in patients studied and secondly due to use of immunoperoxidase technique for the detection of antiovarian antibody. In our study, although the FSH, LH levels were elevated in POF patients shown in [Table 1], no significant association was found between their levels and presence of antiovarian antibody. In conclusion though the number of cases in our study group is not large enough still the role of autoimmunity can be revealed by the presence of antiovarian antibody, which may be one of the probable cause of premature ovarian failure.
| ¤ Summary|| |
Premature ovarian failure is a syndrome consisting of primary or secondary amenorrhoea, hypergonodotropiremia and hypoestrogenemia in women under the age of 40. An autoimmune mechanism was suggested as possible etiology when Vallolton and Forbes in 196667 found antibodies to the cytoplasm of rabbit ova in 29 of 232 tested sera. Immune mechanism in the pathogenesis of premature ovarian failure (POF) is suggested by association of autoimmune phenomenon with POF in some cases and demostration of circulating antibodies to ovary in serum samples from women with POF.  The incidence of presence of antiovarian antibody of POF patients has been reported earlier. Evidence of autoimmunity is present in 18-92%, of patients with POF.  In the present study we have studied 18 cases of POF wits out any overt manifestation of autoimmune disorder but the anti: ovarian antibody was detected, with the idea that this auoantibody might be the cause of ovarian dysfunction which is evident in POF. Presence of antiovarian antibody in 16.67%, cases with POF in our study that ovarian antibodies may play a role in or reflect an autoimmune process responsible for the development of POF.
| ¤ References|| |
|1.||Coulam CB, Ryn RJ. Premature Menopause Etiology, Am J Obstet Gynecol 1979;133:639-42. |
|2.||Alper MM, Jolly F.F,, Garner PR. Pregnancies after premature ovarian failure, Obstet Gynecol 1985; 67:59:61. |
|3.||Irvine WJ, Chan MMV, Scarth L Immunological aspects of premature ovarian failure associated with addison disease. Lancet 1968;II: 883-887. |
|4.||Vasquez AM, Kenny FM. Ovarian failure and antiovarian antibodies in association with hypoparathyroidism, moniliasis and Addison's and Hashimoto's disease. Obsetet Gynecol 1973;41:414-418. |
|5.||Vaidya RA, Aloorkar SD, Reje NR. Premature ovarian failure. T Reprod Med 1977;19:348-351. |
|6.||Damewood MD, Zacur HA, Hoffman G. Circulating antibodies in premature ovarian failure. Obstet Gynecol 1986;71:622-628. |
|7.||Pekonen F, Sieberg R, Makinen T. Immunological disturbances in patients premature ovarian failure. C7in Endocrinal 1986:25:1. |
|8.||Ho PC. Tany GW, Fu KH, Fan MC, Lawton TW. Immunological studies in patients with premature ovarian failure, Obstet Gynecol 1988;71: 622-625. |
|9.||Elder M, Maclaren N, Rilay W. Gonadal autoantibodies in patients with hypogonadism and/or Addison's disease. J Clin Endocrinol 1981;52:1137-1142. |
|10.||Williamson HO, Pharse SA, Mathur RS, Baker ER, Rundenberg AH. Myathema gravis : Premature menopause and thyroid autoimmunity. Am J Obstet Gynecol 1980): 137:893-896. |
|11.||Mignot HM Schoemaker J, Kleingeld M, Ramnath Rao B, Orexhage HA. Premature ovarian failure : The association with antiommunity. Europ J Obst Gynecol Reprobiology, 1989;3-:59-66. |
|12.||Valloton MB, Forbes AP. Immunological disturbances in patients premature ovarian failure. Lancet 1966;2:269-271. |
|13.||Coulam CB. Autoimmune ovarian failure. Semin Report Ehdcrino 1983:1:161-164. |
|14.||Anderson JA, Gondie RB, Gry K, Smith DA. Immunological features of idiopathic Addison's disease : An antibody to cells producing steroid hormone. Clin Exp Immunol 1968;3:107-110. |
|This article has been cited by|
||Premature ovarian failure | [Insuficiencia ovárica prematura]
| ||Scucces, M. |
| ||Revista de Obstetricia y Ginecologia de Venezuela. 2008; 68(4): 263-276 |
||Local lymphocytic and epithelial activation in a case of autoimmune oophoritis
| ||Bats, A.-S., Barbarino, P.M., Bene, M.-C., Faure, G.C., Forges, T. |
| ||Fertility and Sterility. 2008; 90(3): 849,e5-849,e8 |