Peritoneal fluid and serum autoantibody levels in patients with endometriosis

Patients with endometriosis demonstrate autoantibody abnormalities in their peripheral blood. Whether such abnormalities can also be detected in the peritoneal cavity, has not been established. We therefore investigated autoantibodies to 6 phospholipid antigens, 5 histones and histone subfractions and 4 polynucleotides in 14 laparoscopically diagnosed endometriosis and 9 control patients, undergoing laparoscopic tubal occlusion, in both serum and peritoneal fluid. Endometriosis patients demonstrated significantly lower total immunoglobulin G (IgG) levels in peritoneal fluid than controls. In contrast, specific IgG autoantibody levels were higher in peritoneal fluids of endometriosis patients. Mean antiphospholipids and antihistones autoantibodies reached significance when serum/peritoneal fluid ratios were compared with controls. Significant differences between endometriosis and control patients were restricted to IgG isotypes and were not observed for either IgM or IgA isotypes. These data suggest an abnormal concentration of IgG antiphospholipids and antihistones antibodies within the peritoneal cavity of endometriosis patients, which may play a contributing role in the peritoneal pathology of this condition.     

Peritoneal fluid plasminogen activator activity in endometriosis and pelvic adhesive disease

Plasminogen activator activity in PF was assayed by the fibrinolysis method described by Strickland and Beers. In 45 patients studied, there were no discernible differences according to whether patients had endometriosis and/or pelvic adhesive disease. No differences were detected according to when in the menstrual cycle the sample of PF was obtained. These data are in concordance with a previous report and taken together suggest that there is no difference in fibrinolytic mechanisms in PF in patients with or without endometriosis and/or pelvic adhesive disease, when compared with control subjects. If such differences exist, they may be present in the tissues, per se, but are not discernible in PF.     

Association of interleukin-6 and estradiol with hepatocyte growth factor in peritoneal fluid of women with endometriosis

BACKGROUND: Different cytokines and ovarian steroid hormones have been reported to regulate the growth and maintenance of endometriosis. We determined the relationship between peritoneal fluid concentrations of interleukin-6, ovarian steroids and hepatocyte growth factor in different revised American Fertility Society (AFS) staging and morphologic appearances of endometriosis. METHODS: Peritoneal fluid was collected from 30 women with endometriosis and 20 women without endometriosis during laparoscopy, and hepatocyte growth factor, interleukin(IL)-6 and ovarian steroids were measured in peritoneal fluid. The concentrations of hepatocyte growth factor and IL-6 in peritoneal fluid were measured by ELISA, and that of estradiol and progesterone by using the immulyze-enzyme amplified luminescence system. Changes in peritoneal fluid concentrations of hepatocyte growth factor, IL-6, estradiol and progesterone in different stages and morphologic appearances of endometriosis were examined to demonstrate their differences in early and advanced endometriosis. RESULTS: Peritoneal fluid levels of hepatocyte growth factor in women with stage I-II endometriosis were significantly higher than in both women with stage III-IV endometriosis and without endometriosis. A similar significant increase in stage I-II endometriosis was also observed for IL-6 and estradiol. When we divided the women according to different morphologic appearances of endometriosis, we found significantly higher concentrations of hepatocyte growth factor (HGF), IL-6, estradiol and progesterone in women containing red lesions compared with other pigments or without endometriosis. A positive correlation was observed between peritoneal fluid levels of IL-6 and hepatocyte growth factor only but not between other markers. Although estradiol levels in peritoneal fluid showed an increased tendency to elevate in the proliferative phase of endometriosis women, hepatocyte growth factor and progesterone displayed higher concentrations in the secretory phase of the menstrual cycle. After adjusting different variables in peritoneal fluid, multiple analysis of covariance indicated that hepatocyte growth factor levels in peritoneal fluid were independently involved in the red morphologic activity of endometriosis. CONCLUSIONS: Early staging and red color appearance of endometriosis are associated with the elevation in peritoneal fluid concentrations of hepatocyte growth factor, IL-6 and estradiol, demonstrating the combined effect of these cytokines and ovarian steroids in the production of hepatocyte growth factor from endometrial tissues in active endometriosis.     

Vascular endothelial growth factor concentrations in the serum and peritoneal fluid of women with endometriosis.

OBJECTIVES: To investigate whether there is an association between vascular endothelial growth factor (VEGF) levels in serum and peritoneal fluid, and the presence of pelvic endometriosis and its clinical symptoms. METHODS: Blood and peritoneal fluid sample levels of VEGF were measured in 46 women undergoing laparoscopy: 32 with suspected endometriosis and 14 with confirmed endometriosis. Data were analyzed according to phase of the menstrual cycle, symptoms, disease stage, and disease site. RESULTS: There were no significant associations between serum and peritoneal fluid levels of VEGF and the presence of endometriosis, even when controlling for the menstrual phase. However, among the women with confirmed endometriosis, there was a significant increase (P=0.002) in the mean peritoneal VEGF level in those in the late secretory phase compared with those in the proliferative and early secretory phases. CONCLUSIONS: Measuring VEGF levels in symptomatic patients is not helpful to differentiate those with endometriosis from those with a different condition. However, in the late secretory and menstrual phases, mean VEGF levels were higher in women with confirmed endometriosis than in those suspected of having the disease.     

Peritoneal fluid volume, estrogen, progesterone, prostaglandin, and epidermal growth factor concentrations in patients with and without endometriosis

Elevated prostaglandin (PG) levels in peritoneal fluid have been implicated as playing a role in infertility associated with endometriosis. This study was designed to measure peritoneal fluid levels of PG and other hormones that may influence PG release. Specific hormones measured included PGF2 alpha, PGE2, TxB2, 6-keto-PGF1 alpha, estrogen, progesterone, and epidermal growth factor. Peritoneal fluid volume and levels of estrogen, progesterone, and epidermal growth factor were significantly (P less than .05) increased during the secretory, as opposed to the proliferative, phase in both groups of patients, but no significant differences in these parameters were found between patients with and without endometriosis during either the proliferative or secretory phases. Although PG levels did not vary during the menstrual cycle in either group of patients, all four prostanoids were present in significantly (P less than .05) higher concentrations in patients with endometriosis as compared with patients without endometriosis. Furthermore, increased PG levels in patients with endometriosis appear to be due primarily to an increase in PG levels during the secretory phase of the cycle.     

Expression of serum human leukocyte antigen and growth factor levels in a Greek family with familial endometriosis

OBJECTIVE: An increased incidence of endometriosis in the first-degree relatives of patients with endometriosis has been reported, suggesting a familial predisposition and possible genetic influence. In this study, we present a family with four members who have histologically proven endometriosis (mother and three daughters) in two generations and one member with suspected endometriosis in the third generation. The aim of this study was to investigate the presence of serum-soluble class I and class II human leukocyte antigen (sHLA) levels, because they have been shown to be reduced in women with endometriosis. We also studied the levels of vascular endothelial growth factor (VEGF) and epidermal growth factor-receptor (EGF-Rc) whose function in angiogenesis implies an active role in endometriosis. METHODS: Apart from the family members under study, the control groups consisted of 38 women with endometriosis and 30 without any pelvic disease. All the soluble factors under investigation were measured by an enzyme-linked immunosorbent assay technique using a specific immunoassay. RESULTS: All the affected family members and the 38 women with endometriosis had very low levels of serum-soluble class I and class II HLA levels compared with healthy subjects. The circulating levels of VEGF were higher in the endometriosis group than the healthy control group, a pattern in accordance with the family members. On the contrary, EGF-Rc was negative in controls and women with endometriosis, with the exception of certain family members in specific stages of endometriosis. CONCLUSION: We studied the association of endometriosis with circulating levels of human leukocyte antigens and VEGF in two generations of a single family (mother and three daughters). These markers were expressed distinctly in women with familial endometriosis.     

Behaviour of cytokine levels in serum and peritoneal fluid of women with endometriosis

Endometriosis is a disorder characterised by presence and growth of endometrial tissue outside the uterus, primarily into the peritoneum. The peritoneal fluid (PF) of women with endometriosis undergoes a number of biological changes, including local inflammatory-reparative phenomena and peripheral blood mononuclear cells (PBMC) involvement. These activated cells as well as the endometriotic cells secrete various cytokines with pleiotropic biological activities. Dynamic interplay among cytokines may contribute to realise a favourable microenvironment for the implantation of endometrial cells and the progression of the disease. In the present study, we evaluated the levels of cytokines, such as the tumour necrosis factor-alpha (TNF-alpha), transforming growth factor-beta (TGF-beta), interleukin-8 (IL-8) and monocyte chemotactic protein-1 (MCP-1) in PF and in serum (S) of women with endometriosis to compare their behaviour in both biological fluids. The patients (n = 26) were women of reproductive age attending our observation centre for infertility, diagnosed endometriosis at laparoscopy. Control group (n = 5) consisted of women affected by non-immunologic infertility, diagnosed by explorative laparoscopy. S samples were obtained from peripheral blood before anaesthesia and laparoscopy. PF samples were collected at the time of laparoscopy. Both biological fluids were examined for cytokine by ELISA assays. Our results showed that S and PF levels of TNF-alpha, not dosable in controls, were very high at the early stage and decreased significantly with the severity of the disease (p < 0.001). TGF-beta levels were significantly (p < 0.001) higher than in controls and increased with the severity of the disease (p < 0.001), particularly in the PF. S and PF IL-8 as well as MCP-1 concentrations at all stages were higher than in controls (p < 0.001), yet showed an opposite behaviour in both biological fluids. In fact, S levels of IL-8 and MCP-1 were significantly (p < 0.001) higher at early stages and decreased with the severity of the disease, whereas we observed a significant (p < 0.001) enhancement of these chemokine levels in PF from stage I to stage II and stage III. These observations showed that TNF-alpha and TGF-beta levels were overlapping in S and PF of women with endometriosis. On the contrary, MCP-1 and IL-8 S concentrations decreased with the severity of the disease, whereas PF levels showed markedly increased at severe stages. Taken together the observed changes may be due both to the increased peritoneal macrophage activity and to the larger recruitment of PBMC and autocrine release by endometriotic cells.     

Peritoneal fluid fractions from patients with endometriosis do not promote two-cell mouse embryo growth

Peritoneal fluid (PF) from 10 infertile patients with endometriosis, obtained during the follicular phase of the cycle during laparoscopy, did not promote two-cell mouse embryo growth to the extent observed by fluid obtained from seven normal controls. Five molecular weight (MW) fractions were obtained by ultrafiltration, and each was used as media supplement in the assay and compared with PF fractions from normal controls. All fractions of PF from patients with endometriosis inhibited mouse embryo growth to a greater extent than did normal controls. However, the MW fractions greater than 100,000 daltons showed greater inhibition of embryo development than did fractions less than 100,000 daltons. This study of cell-free PF suggests the presence of a humoral factor greater than 100,000 daltons that is inhibitory on mouse embryo growth.     

Overexpression of nerve growth factor in peritoneal fluid from women with endometriosis may promote neurite outgrowth in endometriotic lesions

To investigate the role of the nerve growth factor (NGF) in the endometriosis-associated innervation in the development of endometriosis-associated symptoms, 41 peritoneal fluid samples (PF) from patients with surgically and histologically proven endometriosis and 20 PF from patients with other gynecologic conditions were analyzed with Western blot and a novel in vitro model using dorsal root ganglia (DRG) to show neuronal outgrowth; endometrial cells also were analyzed. The results suggest that the PF of endometriosis patients and endometriotic lesions have neurotropic properties, because the Western blot analysis and the cell culture stainings showed NGF expression, and the neurite outgrowth of DRG treated with PF of patients with endometriosis was significantly higher than when treated with PF of patients without endometriosis. Furthermore, blocking NGF with both anti-NGF and K252a leads to a significant decrease in neurite outgrowth.     

Peritoneal fluid leptin levels are increased but adiponectin levels are not changed in infertile patients with pelvic endometriosis

Objective.?Endometriosis is a leading cause of infertility, and recent studies suggest that leptin and adiponectin may have a role in its causation and progression. This study assessed levels of leptin and adiponectin in serum and peritoneal fluid (PF) in patients with endometriosis and infertility.

Design and setting.?This cross-sectional study included women undergoing diagnostic and/or therapeutic laparoscopy for endometriosis with chief complaint of infertility. Following laparoscopy, patients diagnosed with endometriosis served as cases while patients with no endometriosis served as controls. Patients with polycystic ovarian syndrome, diabetes, thyroiditis and patients on prior therapy with danazol or leuprolide were excluded from the study. Leptin and adiponectin levels were analysed in blood and PF using commercially available ELISA kits.

Results.?Of the 50 patients (aged 22–41 years), 15 had endometriosis (cases) while 35 had no endometriosis (controls). The median PF leptin level was significantly higher in cases as compared to controls (27.7 vs. 15.6?ng/ml, p?=?0.019), and this remained significant even when PF leptin was BMI-normalised (p?=?0.004). However, median serum leptin and adiponectin levels remained comparable between the two groups.

Conclusions.?This study confirmed the role of PF leptin in causation and progression of endometriosis. However, this would have been definitive if healthy fertile females were included in this study.     

Reduced levels of serum pigment epithelium-derived factor in women with endometriosis

The authors previously demonstrated decreased levels of pigment epithelium-derived factor (PEDF) in peritoneal fluid of women with endometriosis compared to women without endometriosis. Here, the authors determine whether women with endometriosis have altered levels of PEDF in serum. Peripheral blood samples were collected from 71 women with and without endometriosis (n = 43 and 28, respectively) before laparoscopic surgery. Concentrations of serum PEDF were measured by enzyme-linked immunosorbent assay. We detected lower levels of serum PEDF in women with endometriosis (16.3 ± 6.6 ng/mL) than in those without endometriosis (24.5 ± 7.3 ng/mL; P < .001). In women with endometriosis, the concentrations of serum PEDF were significantly lower in women with pain (n = 11, 12.6 ± 7.1 ng/mL) compared to women without pain (n = 32, 17.5 ± 6.0 ng/mL; P < .05). However, the concentrations of serum PEDF did not correlate with disease stage or site or infertility. In addition, the concentrations of serum PEDF did not show any difference in the phase of the cycle in either group. Our results suggest that reduced levels of serum PEDF in women with endometriosis and disease-related pain may play a role in the pathogenesis of this disease.     

子宫内膜异位症患者腹腔液肿瘤坏死因子与精子活动力的关系

本研究用双抗体夹心法，检测１６例子宫内膜异位症不孕妇女（异位症组）腹腔液肿瘤坏死因子（ＴＮＦ）含量，及其对精子活动能力和质膜完整性的影响，并以１１例其他病因所致不孕妇女和７例正常妇女为对照组进行比较。结果表明，异位症组ＴＮＦ含量明显增高，且重度患者含量高于轻度患者；异位症组腹腔液与供精者精子共同孵育后，后者精子前向运动率、总活动率和低渗肿胀率均显著降低，并与ＴＮＦ水平呈明显负相关。提示：异位症患者腹腔液ＴＮＦ水平增高对精子有抑制作用，可能是导致不孕的因素之一。     

Association between endometriosis and polymorphisms in endostatin and vascular endothelial growth factor and their serum levels in Korean women

The aim of present study was to evaluate the relationship between endometriosis and polymorphisms in the endostatin and vascular endothelial growth factor (VEGF) genes, and their levels in serum in a Korean population. Serum endostatin levels (but not VEGF levels) were negatively correlated with the development of endometriosis, specifically in early-stage endometriosis patients, compared with women without endometriosis, but endometriosis was not associated with the endostatin G(4349)A and VEGF C(936)T polymorphisms.     

Peritoneal fluid concentrations of interleukin-11 in women with endometriosis

Objective: To determine the peritoneal fluid concentrations of interleukin-11 (IL-11) in women with endometriosis as compared with the control group.

Design: A prospective, controlled study.

Setting: The obstetrics and gynecology department of a teaching hospital and a university immunology department.

Patient(s): Sixty consecutive women undergoing laparoscopic surgery for benign gynecological indications.

Intervention(s): Peritoneal fluid was obtained during laparoscopy, and the concentration of IL-11 was measured.

Main Outcome Measure(s): Concentration of IL-11 in correlation with the presence of endometriosis, its stage, and the phase of the menstrual cycle.

Result(s): IL-11 was detectable in the peritoneal fluid of 64% of women tested. Concentrations of IL-11 showed no correlation with the presence of endometriosis, the American Fertility Society stage of the disease, or the phase of the menstrual cycle.

Conclusion(s): We found no evidence to suggest that IL-11 is involved in the pathogenesis of pelvic endometriosis.     

The evaluation of IL-12 levels in peritoneal fluid and serum of women with endometriosis

We studied levels of IL-12 in peritoneal fluid and serum in patients with minimal, advanced and recurrent endometriosis compared to women without endometriotic lesions in pelvis minor. The aim of the study was to determine whether level of IL-12 detected in peritoneal fluid or serum changes with grading of severity of endometriosis. To assess IL-12 levels immunosorbent ELISA was used. There were no statistically significant differences in IL-12 levels in peritoneal fluid nor in serum in any of studied groups. There was higher concentration (without statistical significance) of IL-12 in peritoneal fluid of healthy women compared to women with endometriosis.     

Peritoneal fluid concentrations of ciliary neurotrophic factor,a gp130 cytokine,in women with endometriosis

Ciliary neurotrophic factor (CNTF) is a gp130 neuroregulatory cytokine belonging to the interleukin-6-type cytokine superfamily. Several lines of evidence suggest that the concentrations of interleukin-6 are elevated in the peritoneal fluid of endometriosis patients. To our knowledge, no study on whether this might also be the case for CNTF levels has been published previously. The CNTF concentrations in the peritoneal fluid were measured by ELISA in 51 women with (n = 31) and without (n = 20) endometriosis. Surgery was scheduled during the proliferative or the secretory phase of the menstrual cycle. CNTF was detectable in the peritoneal fluid of 43% of the women tested. The concentrations of CNTF showed no correlation with the presence of endometriosis or the phase of the menstrual cycle. We found no evidence to suggest that CNTF is involved in the pathogenesis of pelvic endometriosis.     

Peritoneal fluid and serum steroids in infertility patients

Peritoneal fluid and serum were collected from 78 patients at the time of laparoscopy. Twenty-two were fertile controls (CTL), and 56 were infertility patients, who were subdivided into three main groups: endometriosis (EMS), pelvic adhesions (ADH), and ovarian dysfunction (OvDF). Based on control group data, biochemical criteria indicative of the presence of a stigma, S(+), were established: (1) serum progesterone (P) greater than or equal to 2 ng/ml, (2) peritoneal fluid P greater than or equal to 50 ng/ml, and (3) peritoneal fluid/serum ratio of P greater than or equal to 3. Direct visualization by laparoscopy showed that 21% CTL, 75% EMS, 69% ADH, and 56% OvDF subjects had luteinized unruptured follicle (LUF) syndrome. Biochemical criteria, however, demonstrated only 7% CTL, 37% EMS, 23% ADH, and 56% OvDF subjects had LUF. Peritoneal fluid estradiol (E2) and P concentrations and total content were significantly lower in LUF than in non-LUF patients, whereas serum E2 and P concentrations were not different between the two groups. Values for testosterone and androstenedione in peritoneal fluid and serum were similar between these two groups. Endometrial dating in LUF versus non-LUF patients were also similar. The usual indicators of ovulation, i.e., serum P, endometrial dating, and basal body temperature, failed to identify LUF. The diagnosis of LUF can be best made by P assay of peritoneal fluid and serum.