Inflammation related to high-mobility group box-1 in endometrial ovarian cyst.

Endometriosis is a chronic inflammatory disease often associated with dysmenorrhea, infertility, adenomyosis, and endometrial ovarian cyst (EOC). In particular, EOC can sometimes become malignant in a longitudinal follow-up. This study aimed to investigate the involvement of high-mobility group box-1 (HMGB1) in an inflammatory milieu and the characteristics of immune cells in EOC. The samples were obtained from patients who underwent ovarian cystectomy for benign ovarian cyst. The participants were divided into two groups: patients with EOC (EOC group) and those without EOC (nEOC group). We divided a part of the removed ovary into small sections and isolated the tissue cells. Thereafter, the cytoplasmic HMGB1 levels in DCs, macrophages, and non-immune cells were analyzed by flow cytometry. We also evaluated the proportions of immune, T, NK, iNKT, NK, and regulatory T (Treg) cells. Results showed that the DCs, macrophages, and non-immune cells of EOC had significantly higher cytoplasmic HMGB1 levels than those of nEOC. The expression of CD69 and CD107a on CD8+ T and CD4+ T cells of EOC was also more enhanced than that of nEOC. Furthermore, the M2 macrophages and Tregs highly accumulated in EOC. These results indicate that HMGB1 may aggravate chronic inflammation related to T-cell activation and simultaneously facilitate development of the immunosuppressive milieu in EOCs.

Impact of lipopolysaccharide administration on luteinizing hormone/choriogonadotropin receptor (Lhcgr) expression in mouse ovaries.

Lipopolysaccharide (LPS) is isolated from the genital tract of animals suffering from uterine damage and ovarian dysfunction. This study provides direct molecular evidence about the mechanism through which endotoxins cause reproductive disorders. Granulosa cells and ovaries were collected from immature mice treated with eCG or with eCG and LPS injection intraperitoneally. Normal large antral follicles were observed in ovaries obtained from eCG and LPS coinjected mice, and the morphology of the ovaries was similar to that observed in the control group. These antral follicles were not deemed atretic because few TUNEL-positive cells were observed. However, the granulosa cells of large antral follicles did not acquire the ability to respond to hCG stimulation. The number of ovulated oocytes was significantly lower in LPS-injected mice after superovulation compared to mice that were not exposed to LPS. The low reactivity was caused by the limited expression of the Lhcgr gene, which encodes the LH receptor in granulosa cells as well as an LPS-induced increase in the level of Dnmt1 expression. The methylation rate of the Lhcgr promoter region was significantly higher in granulosa cells obtained from the LPS treatment group compared with the control group. Together, these findings demonstrated that the decrease in the expression of Lhcgr due to LPS was a result of the epigenetic regulatory action of LPS. Our studies suggest that ovarian follicular cysts that is characterized by bacterial infection in humans and animals, is closely connected to the level of methylation of the Lhcgr promoter region.

NATURALLY-OCCURRING AUTOANTIBODIES TO HUMAN CHORIONIC GONADOTROPIN AND ITS SUBUNITS IN OVARIAN CYST PATIENTS.

Growing evidence supports the existence of immune-surveillance mechanisms in ovarian tumour patients, including autoantibodies to tumour associated and tumour specific antigens, tumour growth factors. Glycoprotein hormone human chorionic gonadotropin (hCG) and its hormone-specific hCGß subunit have been associated with epithelial tumours such as bladder, lung, oral/facial, breast, cervical, ovarian, vaginal, prostate, renal and pancreatic carcinomas. It is believed that hCG plays a role of an autocrine growth factor for tumor cells. Here we have demonstrated that sera of patients with ovarian cyst contain naturally-occurring autoantibodies, predominantly of IgG2 isotype, that bind to hCG and its subunits with high affinity. Titration of blood sera from 36 female patients, aged 22-61 after ethical permission and informed consent, diagnosed with ovarian cyst and healthy age-matched controls (n=12) was performed using a classical enzyme-linked immunosorbent assay (ELISA). Binding of the sera to the following antigens was tested: hCGαß, hCGß, hCGα, hCGß C-terminal peptide (hCGßCTP) and hCGß core fragment (hCGßCF). The same type of ELISA (with necessary modifications) was used for further investigation of subclass usage and assessment of binding affinity of the detected autoantibodies. Our data indicates that the sera of the majority of patients with ovarian cyst contain significantly higher levels of the natural IgG antibodies binding to hCGαß, hCGß, hCGα, hCGßCTP and hCGßCF, than those of the healthy controls. Natural IgG antibodies to hCGαß heterodimer were detected in 78% of cases, to hCGß in 61% of cases, to hCGα in 78% of cases, to hCGßCTP in 69% of cases, to hCGßCF in 83% of cases. These autoantibodies predominantly belonged to the IgG2 subclass and were characterized by the high binding affinity. It is plausible that they cross- bind to sugar side chains of hCG and its subunits. Our data demonstrated that sera of patients with the ovarian cyst contains elevated levels of naturally-occurring IgG antibodies, which bind to hCG and/or its subunits. The overwhelming majority of these autoantibodies belong to the IgG2 isotype thus indicating that they might be directed against carbohydrate antigens within highly glycosylated hCG.

Role of Intracystic Cytokines and Nitric Oxide in Ovarian Neoplasms.

The development of new biomarkers for the diagnosis and prognosis of ovarian cancer may provide an opportunity for new therapies. In this study, we aimed to compare cytokines (interleukin [IL]-2, IL-5, IL-6, IL-8, IL-10 and tumour necrosis factor [TNF]-α) and nitric oxide (NO) metabolite levels in non-neoplastic tumours, benign primary ovarian tumours and malignant primary ovarian neoplasms. The secondary aim was to relate cytokine and intracystic NO metabolite levels to clinical, laboratory and pathologic characteristics for patients with primary ovarian malignancies. We evaluated 110 patients with adnexal masses. Cytokine concentrations were quantified by enzyme-linked immunosorbent assay and nitrate concentrations by enzymatic reduction of nitrite by nitrate reductase. Patients with malignant neoplasms had higher IL-6, IL-8 and NO levels compared to patients with benign neoplasms. Histologic grade 1 tumours were associated with elevated IL-2 levels, whereas anaemia was associated with elevated IL-6 levels. On average, those patients with elevated IL-8 levels also had a neutrophil/lymphocyte ratio (NLR) greater than 2.6 and less than 36 months of disease-free survival (DFS). Patients with normal CA 19-9 levels had elevated IL-10 levels. TNF-α was elevated in patients with two carcinogenesis and those with a platelet/lymphocyte ratio (PLR) less than 300. NO levels were higher in patients with an NLR less than 2.6 and CA 19-9 greater than 35 U/ml. Elevated intracystic cytokine levels, especially IL-6 and IL-8, are associated with worse prognosis in ovarian cancer.

Altered Expression of Pro-inflammatory Cytokines in Ovarian Follicles of Cows with Cystic Ovarian Disease.

A growing body of evidence suggests that ovulation shares many of the features of an inflammatory reaction and that cytokines play many diverse and important roles in reproductive biology. The aim of this study was to examine the expression of the pro-inflammatory cytokines interleukin (IL)-1α, IL-6 and tumour necrosis factor (TNF)-α in ovarian cells from cows with cystic ovarian disease (COD) as compared with that in ovarian structures from regularly cycling cows. Expression of genes encoding IL-1α, IL-6 and TNF-α was detected by real-time polymerase chain reaction in follicular cells from ovaries from healthy cows and cows with COD with no significant differences. However, immunohistochemistry showed increased expression of IL-1α, IL-6 and TNF-α in cystic follicles, suggesting that this expression may be related to the persistence of follicular cysts. The effect of COD was evident for IL-1α and TNF-α, and a follicular structure-disease interaction was observed in the expression of all the cytokines evaluated. Thus, altered expression of these proinflammatory cytokines may be related to ovulation failure and development of follicular cysts.

A tumor necrosis factor-α inhibitor reduces the embryotoxic effects of endometriotic peritoneal fluid.

OBJECTIVE: To determine whether a tumor necrosis factor-α (TNF-α) inhibitor can reduce the embryotoxicity of the peritoneal fluid (PF) of women with endometriosis. DESIGN: Experimental clinical study. SETTING: University hospital. PATIENT(S): Twelve women with chocolate cysts and 12 control women without endometriosis. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): We collected the PF from patients with chocolate cysts (CH-PF) and patients without endometriosis (N-PF) during laparoscopic surgery. For the in vitro studies, development and apoptosis were evaluated in two-cell stage mouse embryos after incubation with CH-PF and N-PF, with or without a TNF-α inhibitor. RESULT(S): We found that CH-PF significantly decreased the rate of blastocyst development and increased the percentage of apoptotic cells in the embryos. Cytokine assays showed that the concentrations of several cytokines, including TNF-α, were higher in embryos incubated with CH-PF than in those incubated with N-PF. Furthermore, the treatment of embryos with TNF-α retarded development and induced apoptosis. Important, adalimumab, a TNF-α inhibitor, effectively abrogated the embryotoxicity that was induced by CH-PF. CONCLUSION(S): These data collectively highlight the crucial role of TNF-α in CH-PF-induced embryotoxicity and suggest that TNF-α inhibitors may be potential therapeutic agents for treating endometriosis-induced infertility.

Immunoexpression of the PTEN protein and matrix metalloproteinase-2 in endometrial cysts, endometrioid and clear cell ovarian cancer.

OBJECTIVES: Endometrioid and clear cell ovarian adenocarcinomas are suspected to derive from ectopic endometrial foci. The aim of the study was to determine PTEN and MMP-2 immunoexpression in endometrial ovarian cysts, endometrioid and clear cell ovarian carcinomas and to assess the relationship between the abovementioned values and clinical data of patients in order to find the marker of increased risk of malignant proliferation based on ovarian endometriotic lesions. Detailed analysis of the collected data was conducted to investigate the correlation between immunohistochemical expression of the examined antigens, histopathological diagnosis and clinical condition of patients. MATERIAL AND METHODS: 20 endometrial adenocarcinomas, 21 clear cell ovarian cancers and 26 endometrial cysts were included in the study The control group consisted of 29 specimens of physiological endometrium: 16 samples of the proliferative phase and 13 samples of the secretory phase. Protein expression of PTEN and MMP-2 was evaluated by immunohistochemistry Protein immunoexpression in the collected specimens was estimated with the use of light microscope and MultiScan software. Immunoreactivity of the PTEN antigen was assessed by the quantitative method, whereas MMP-2 immunoexpression was evaluated by the semi-quantitative method. Two-sided tests were used for statistical inference. Generalized linear models were used to compare the studied groups. Error distributions were selected using the Akaike criterion (AIC). Statistical analysis was conducted with the use of the R Statistical Package. RESULTS: MMP-2 immunoreactivity differed significantly between the study groups and controls (p<0.001). PTEN immunoexpression was the strongest in endometrial cysts (53.7 %), lower in clear cell cancers (50.2%) and the lowest in endometrioid adenocarcinomas (43.88%), but the differences were not statistically significant (p=0.17). PTEN reactivity in the group of endometrioid carcinomas was significantly higher (p=0.02), while MMP-2 expression had a falling tendency (p=0.076) in obese women. CONCLUSIONS: Increased MMP-2 expression in the successive groups may imply a rising invasive potential of the epithelial cells in endometrial cysts, endometrioid and clear cell adenocarcinomas. Strong immunoreactivity for PTEN in proliferative endometrium implies its role in the regulation of endometrial proliferation. PTEN activity may reduce MMP-2 expression in insulin resistant women suffering from endometrial ovarian cancer Simultaneous evaluation of PTEN and MMP-2 immunoexpression in ectopic endometrial foci cannot be used to identify women with an increased risk of neoplastic transformation.

Increased levels of oxidative stress markers in the peritoneal fluid of women with endometriosis.

OBJECTIVE: To evaluate 8-hydroxy-2-deoxyguanosine (8-OHdG) and 8-isoprostane levels in the peritoneal fluid (PF) of women with endometriosis. STUDY DESIGN: One hundred and ten women with laparoscopically and histopathologically confirmed endometriosis and, as reference groups, 119 patients with simple serous (n=78) and dermoid (n=41) ovarian cysts were studied. Peritoneal fluid 8-OHdG and 8-isoprostane concentrations were evaluated by enzyme-linked immunosorbent assays. RESULTS: 8-OHdG and 8-isoprostane levels in peritoneal fluid were significantly higher in patients with endometriosis compared with the reference groups. Higher PF 8-OHdG and 8-isoprostane concentrations were observed in patients with advanced stages of endometriosis. A statistically significant positive correlation was found between 8-OHdG and 8-isoprostane levels in peritoneal fluid. CONCLUSION: Endometriosis induces greater oxidative stress and frequent DNA mutations in peritoneal fluid than nonendometriotic ovarian cysts. The most severe oxidative stress occurs in the peritoneal cavity of women with more advanced stages of the disease.

CD4⁺ CD25⁺ FOXP3⁺ regulatory T cells in peripheral blood and peritoneal fluid of patients with endometriosis.

STUDY QUESTION: Is endometriosis associated with changes in CD4⁺ CD25⁺ FOXP3⁺ regulatory T cells (Treg cells)? SUMMARY ANSWER: Endometriosis is associated with disturbed compartmentalization of CD25(high) FOXP3⁺ Treg cells. WHAT IS KNOWN ALREADY: Endometriosis is associated with an abrogated immune response and displays some features of an autoimmune disorder. Treg cells play a part in the development of autoimmune reactions; however, their role in pathogenesis of endometriosis is still poorly recognized. STUDY DESIGN, SIZE AND DURATION: Case-control study comparing 17 women with laparoscopically and histopathologically confirmed ovarian endometriosis with 15 control women without visible endometriosis foci, pelvic inflammation or related pathology who were subjected to laparoscopic surgery between 2010 and 2011. PARTICIPANTS/MATERIALS, SETTING AND METHODS: Peripheral blood and peritoneal fluid were collected during laparoscopy and T cell subpopulations were analysed by flow cytometry using specific monoclonal antibodies recognizing CD4⁺, CD25⁺ and FOXP3⁺ markers. MAIN RESULTS: The percentage of CD25(high) FOXP3⁺ Treg cells was significantly decreased in the peripheral blood of women with ovarian endometriosis compared with control women. On the other hand, the proportion of these cells was significantly increased in the peritoneal fluid of women with endometriosis. LIMITATIONS, REASONS FOR CAUTION: The present study is limited to patients with ovarian endometrioma and further investigations are needed, including patients with lower grade of endometriosis. WIDER IMPLICATIONS OF THE FINDINGS: The present results suggest that Treg cells may play a part in immunopathogenesis of endometriosis being responsible for abrogated local cellular immune responses and facilitation and development of autoimmune reactions. Treg cells may be thus a potential target in the treatment of endometriosis. STUDY FUNDING/COMPETING INTEREST(S): This study was supported by 1M15/N/2011 and NK1W grants from the I Faculty of Medicine, Warsaw Medical University. None of the authors has any competing interests to declare.

Clinical importance of determination of cytokines in patients with ovary tumours.

Benign ovary tumours share about 75-87% of true ovary tumours. Authentically high indicator of IL-6 and the TNF-alpha have all types of ovary tumours. Benign ovary formations are accompanied by lower maintenance of IL-6 and the TNF-alpha in blood serum, than at malignant tumours.

Esophageal cyst producing CA19-9 and CA125.

The patient was a 59-year-old woman in whom computed tomography revealed a posterior mediastinal cyst and ovarian cystoma at a medical check-up in March 2007. Blood tests showed high CA19-9 and CA125 levels. She underwent left adnexectomy for ovarian cystoma in July 2008 and histopathological examination led to a diagnosis of dermoid cyst. The postoperative levels of CA19-9 and CA125 remained high. She developed dysphagia in February 2009, and the posterior mediastinal cyst showed a tendency to enlarge. Therefore, she underwent tumorectomy through a small thoracotomy. The cyst contained greenish fluid with CA19-9 and CA125 contents of 65,000 and 78,000 U/ml, respectively. Histologically, the cyst had a thickened wall, which contained two muscle layers, and was lined by squamous and pseudostratified ciliated epithelium. No cartilage or bronchial glands were identified. These findings led to a diagnosis of esophageal cyst. On immunohistochemical staining, the cyst-lining epithelial cells were positive for CA19-9 and CA125. The serum CA19-9 and CA125 levels returned to normal two months after surgery. We report a resected case of esophageal cyst producing CA19-9 and CA125.

TNFalpha gene silencing reduced lipopolysaccharide-promoted proliferation of endometriotic stromal cells.

PROBLEM: We previously reported that lipopolysaccharide (LPS)-promoted endometriotic stromal cell (ESC) proliferation by inducing TNFalpha production. The aim of this study was to investigate the efficacy of TNFalpha gene silencing on LPS-treated ESCs. METHOD OF STUDY: Endometriotic stromal cells (ESCs) and endometrial stromal cells (ESCs) (EMSCs) were obtained from ovarian chocolate cysts and uterine myoma, respectively. Using PCR array, LPS-induced gene expression profiling after transfection of TNFalpha siRNA into ESCs was performed. Down-regulated genes by TNFalpha silencing were examined using real-time RT-PCR. Effect of TNFalpha silencing was examined using ELISA and BrdU incorporation, respectively. RESULTS: In PCR array, TNFalpha silencing in ESCs repressed LPS-induced expression of cIAP2 and IL-8, NFkappaB pathway responsive genes. After adding LPS, the levels of cIAP2 and IL-8 expression in ESCs were higher compared with those in EMSCs. TNFalpha silencing attenuated the LPS-induced ESC proliferation. CONCLUSION: Tumor necrosis factor alpha may be involved in cell proliferation of endometriotic tissues.

Expression of sialyl Lex, sialyl Lea, Lex and Ley glycotopes in secreted human ovarian cyst glycoproteins.

Human blood group A, B, H, Ii, Le(a) and Le(b) antigens and their determinants expressed on ovarian cyst glycoproteins have been studied for over five decades. However, little is known about sialyl Le(x) and sialyl Le(a) glycotopes, which play essential roles in normal immunity, inflammation, and cancer cell metastasis. Furthermore, Le(x) and Le(y) were classified as glycotopes of unknown genes. Identification of these Lewis epitopes was hampered by the lack of specific antibodies. In this study, the occurrence of sialyl Le(x), sialyl Le(a), Le(x) and Le(y) reactivities in cyst glycoproteins was characterized by enzyme-linked immunosorbent assays. The results indicated that most human ovarian cyst glycoproteins carried Le(x) (8/25) and/or Le(y) (17/25) glycotopes. The expression (epitopes) of the new genes described in previous reports are Le(x) and Le(y) glycotopes; the reactivities of sialyl Le(x) and sialyl Le(a) glycotopes in secreted cyst glycoproteins may be affected by the conditions of purification; the relationship between Le(y) and human blood group ABH was confirmed; recognition profiles of sialyl Le(x), sialyl Le(a), Le(x) and Le(y) present in the carbohydrate chains of water-soluble cyst glycoproteins were illustrated; possible attachments of glycotopes to the internal carbohydrate complex of cyst glycoproteins have been reconstructed; proposed biosynthetic pathways for the formation of sialyl Le(a), sialyl Le(x), Le(x), Le(y), ALe(y) and BLe(y) determinant structures on Type I and Type II core structures of human ovarian cyst glycoproteins are also included in this study.

Massive ascites as a presentation in a young woman with endometriosis: a case report.

OBJECTIVE: To report a case of endometriosis associated with massive ascites and an elevated CA-125 level. DESIGN: Case report. SETTING: Tertiary care center. PATIENT(S): A 26-year-old woman presented with massive ascites and an increased CA-125 level suggestive of ovarian cancer. INTERVENTION(S): Ultrasonography, laparotomy, and bilateral ovarian cystectomy and reconstruction. Endometriosis was diagnosed postoperatively on the basis of histopathology. The patient received 6 months of treatment with a GnRH analogue. MAIN OUTCOME MEASURE(S): Ultrasound examination 6 months after surgery to evaluate for ascites or recurrent ovarian cysts. RESULT(S): Frozen sections obtained at laparotomy and ovarian cystectomy ruled out a malignancy. The final histologic report was compatible with a diagnosis of endometriosis. After 6 months of treatment with the GnRH analogue, the patient experienced a progressive reduction of the ascitic fluid and full remission after 2 years. CONCLUSION(S): Endometriosis associated with massive bloody ascites is an unusual occurrence. This report draws attention to this condition as a complication of endometriosis. For this reason, endometriosis should be included in the differential diagnosis of reproductive-age women presenting with an apparent ovarian malignancy.

Immunohistochemical evaluation of canine ovarian cysts.

To clarify the immunohistochemical characteristics of canine ovarian cysts, 109 canine ovarian cysts (57 cysts of subsurface epithelial structures: SES, 26 graafian follicle cysts, 12 cystic rete ovarii and 14 cysts difficult to classify morphologically) were examined regarding their lining cells immunohistochemically using antibodies against placental alkaline phosphatase (PLAP), S100, inhibin alpha, desmin and AE1/AE3. Both cysts of SES and cystic rete ovarii had a positive immunoreaction to desmin and AE1/AE3, whereas all cysts all but graafian follicle cysts were negative for inhibin alpha. PLAP-positive immunoreaction was observed only in cysts of SES. Graafian follicle cysts had a positive immunoreaction to inhibin alpha, but were negative for PLAP, desmin and AE1/AE3. Fourteen cysts were difficult to classify morphologically because these cysts had single-squamous lining cells and lacked other morphological characteristics. However, these unclassified cysts were immunohistochemically divided into two groups, including positive and negative cysts, by the reactivity of PLAP. The PLAP-positive cysts were considered large cysts of SES. These results suggest that PLAP was a useful marker for classification of cysts of SES, although cysts originating from SES are not always positive for this antigen.

Macrophages in human fallopian tube and ovarian epithelial inclusion cysts.

Epithelial inclusion cysts (EICs) are considered a preferential site for ovarian carcinogenesis. Local inflammation, associated to ovulatory wound repair and epithelial inflammatory conditions, facilitates EIC formation and involves activation of macrophages. The aim of this study was to analyse the presence and numbers of macrophages in the ovarian surface epithelium (OSE), in EICs, and in the fallopian tubes, as tubal metaplasia is a common finding in EICs. Immunohistochemical analysis of macrophages was performed in 25 fallopian tubes in different phases of the menstrual cycle, and in 30 ovaries showing EICs from cycling and postmenopausal women. In the fallopian tube, macrophages were abundant and underwent cyclic changes during the menstrual cycle, being particularly abundant within the epithelium at early and mid-luteal phases. Macrophages were not found in the normal OSE. However, OSE areas and EICs showing tubal metaplasia were invariably associated with infiltration by abundant macrophages. Macrophages were present among epithelial cells, infiltrating the cyst wall, as well as free in the cyst lumen. No significant differences existed between follicular and luteal phases of the cycle, or between cycling and postmenopausal women. This study has demonstrated that macrophages are associated with metaplastic EICs, and raises the possibility that these cells contribute to the particular microenvironment of EICs through secretion of cytokines and growth factors that may reach bioactive concentrations in the confined space of the EICs.

Laparoscopic excision of ovarian cysts does not result in antiovarian humoral autoimmunity.

To determine whether the ovarian trauma consequent to the laparoscopic removal of a cyst could result in the development of a humoral immunity, antiovarian antibodies were assayed in serum samples obtained from 40 women before and after cystectomy.

[Assessment of soluble intracellular adhesion molecule-1 (sICAM-1) in women with benign ovarian tumors]. / Ocena stezen rozpuszczalnych miedzykomórkowych molekul adhezyjnych typu 1 (sICAM-1) u kobiet z guzami niezlosliwymi jajników.

OBJECTIVES: Overexpression of intracellular adhesion molecule-I (ICAM-1) was observed in many benign and malignant tumors. The aim of our study was to evaluate its serum concentrations as well as CA-125 in women with benign ovarian tumors. MATERIALS AND METHODS: Forty-five women treated surgically because of benign ovarian mass. RESULTS: Mean concentrations of sICAM-1 in benign tumors was 241.8+/-74.1 ng/ml and 195.6+/-68.7 ng/ml in healthy controls. No correlations between sICAM-1 concentrations and leukocyte count, tumor volume, BMI and obstetrical history. Efficiency in tumor differentiation was higher for CA-125 than sICAM-1 (area under Receiver Operating Characteristic curve 0.78 and 0.63 respectively). We observed higher sICAM-1 concentrations in fibrothecomas and lower in endometrial and dermoid cysts. CONCLUSIONS: Serum ICAM-1 concentrations correlate with some histological types of benign tumors, but not with tumor volume. Levels of CA-125 are more effective than ICAM-1 in ovarian tumors differentiation.

Effects of dehydroepiandrosterone on ovarian cystogenesis and immune function.

The purpose of the present report was to study the possible relationship between ovarian functionality and the immune response during cystogenesis induced by androgenization with dehydroepiandrosterone (DHEA). Daily injection of DHEA (6 mg/kg body weight) for 20 consecutive days induced ovarian cysts in BALB/c mice. As markers of ovarian function, serum estradiol (E) and progesterone (P) and the ovarian inmunomodulator prostaglandin E (PGE) were analyzed. In order to know how the integrity of the tissue was altered after induction of cystogenesis, the oxidative status was also evaluated. Serum E and P levels, and ovarian PGE concentration, were increased in animals with cysts compared with healthy controls. The oxidant status (quantified by malondialdehyde (MDA) formed after the breakdown of the cellular membrane by free radical mechanisms) was augmented, meanwhile the antioxidant (evaluated by the glutathione (GSH) content) diminished during the induction of cystogenesis. Both immunohistochemical and flow cytometry assays demonstrated that DHEA treatment increased the number of T lymphocytes infiltrating ovarian tissue. Therefore, while ovarian controls showed equivalent expression of CD4+ and CD8+ T cell subsets, injection of DHEA yielded a selective ovarian T cell infiltration as demonstrated by enhanced CD8+ and diminished CD4+ T lymphocyte expression. These results show that the development of cysts involves changes in ovarian function and an imbalance in the oxidant-antioxidant equilibrium. We observed also both an increased and selective T lymphocyte infiltration.