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Year : 1999  |  Volume : 53  |  Issue : 6  |  Page : 254-258

Antiovarian antibody in premature ovarian failure

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

Correspondence Address:
D Chattopadhyay
Department of Immunology, Institute of Medical Sciences, Banares Hindu University, Varanasi - 221 005.
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Source of Support: None, Conflict of Interest: None

PMID: 10776506

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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

How to cite this URL:
Chattopadhyay D, Sen M R, Katiyar P, Pandey L K. Antiovarian antibody in premature ovarian failure. Indian J Med Sci [serial online] 1999 [cited 2016 May 31];53:254-8. Available from:

 ¤ Introduction Top

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. [1],[2] POF is associated with a variety of autoimmune disorders such as Addison's disease, [3] thyroid disorder. [4],[5] Works of these authors have shown antiovarian autoanti­boies presence in women with pre­mature ovarian failure. [6],[7],[8] Ovarian failure occurs in high proportion of women with autoimmune disea­ses [9] The demonstration of circulat­ing antibodies to ovarian tissue of women with premature ovarian fai­lure was explained. [10] 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. [6] 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. [6] 24 patients positive for non-organ specific and organ specific autoantibodies are studied in order to establish a relationship between POF and autosensitiza­tion. [11] In our present study the presence of antiovarian antibody in the test sera was detected by immunoperoxidase technique [6] with slight modification.

 ¤ Material and Method Top

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 Top

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 Top

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. [2],[ 7] Evidence of autoimmunity is present in 12-18% of patients with POF. [8],[11] 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 Top

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. [13] 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. [14] 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 Top

1.Coulam CB, Ryn RJ. Premature Menopause Etiology, Am J Obstet Gynecol 1979;133:639-42.  Back to cited text no. 1    
2.Alper MM, Jolly F.F,, Garner PR. Pregnancies after premature ova­rian failure, Obstet Gynecol 1985; 67:59:61.  Back to cited text no. 2    
3.Irvine WJ, Chan MMV, Scarth L Immunological aspects of prema­ture ovarian failure associated with addison disease. Lancet 1968;II: 883-887.  Back to cited text no. 3    
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.  Back to cited text no. 4    
5.Vaidya RA, Aloorkar SD, Reje NR. Premature ovarian failure. T Re­prod Med 1977;19:348-351.  Back to cited text no. 5    
6.Damewood MD, Zacur HA, Hoffman G. Circulating antibodies in prema­ture ovarian failure. Obstet Gyne­col 1986;71:622-628.  Back to cited text no. 6    
7.Pekonen F, Sieberg R, Makinen T. Immunological disturbances in pa­tients premature ovarian failure. C7in Endocrinal 1986:25:1.  Back to cited text no. 7    
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.  Back to cited text no. 8    
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.  Back to cited text no. 9    
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.  Back to cited text no. 10    
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.  Back to cited text no. 11    
12.Valloton MB, Forbes AP. Immuno­logical disturbances in patients pre­mature ovarian failure. Lancet 1966;2:269-271.  Back to cited text no. 12    
13.Coulam CB. Autoimmune ovarian failure. Semin Report Ehdcrino 1983:1:161-164.  Back to cited text no. 13    
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.  Back to cited text no. 14    


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