Cytokine profiles in serum and peritoneal fluid from infertile women with and without endometriosis

OBJECTIVE: To study the serum and peritoneal fluid cytokine profiles in infertile women with minimal/mild active endometriosis. METHODS: Fifty-seven consecutive infertile women undergoing laparoscopy for unexplained infertility had peritoneal fluid and serum samples obtained at the time of laparoscopy. The levels of interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-1 beta (IL-1 beta), vascular endothelial growth factor (VEGF), tumor necrosis factor-alpha (TNF-alpha), monocyte chemotatic protein-1 (MCP-1), RANTES, platelet derived growth factor (PDGF), soluble Fas (sFas), and soluble Fas Ligand (sFasL) in peritoneal fluid and serum were measured to compare the concentration in both biological fluids, in women who have minimal/mild red endometriosis using women with no endometriosis as controls. RESULTS: Peritoneal fluid levels of MCP-1, IL-8 and IL-6 were significantly higher in the endometriosis group (P < 0.012, P = 0.003, and P = 0.015, respectively). There was no significant difference in the peritoneal fluid levels of IL-1 beta, TNF-alpha, RANTES, VEGF, PDGF, sFas and sFasL in the two groups. Although serum levels of IL-8 were higher in women with endometriosis, the difference was not significant (P = 0.07). Serum levels of PDGF, IL-6, RANTES, IL-1 beta, TNF-alpha, and sFas, were not significantly different in the two groups. CONCLUSION: The elevated levels of MCP-1, IL-6, and IL-8 in peritoneal fluid but not serum may indicate the importance of local macrophage activating factors in the pathogenesis of endometriosis.     

Polymorphism of the interleukin-1beta gene and endometriosis

OBJECTIVE: Experimental data indicate that interleukin (IL)-1 plays a role in the pathogenesis of endometriosis. We determined the genotype and the allele frequencies of the IL1B exon 5 polymorphism and the corresponding IL-1beta serum levels in women with endometriosis. METHODS: We genotyped 92 women with surgically and histologically confirmed endometriosis and 69 controls without history of endometriosis. Both groups were of middle European genetic background for the IL1B exon 5 polymorphism (at position +3954). Genotyping was performed by pyrosequencing. IL-1beta serum levels were analyzed by a commercially available enzyme-linked immunosorbent assay. RESULTS: Allele frequencies in women with endometriosis and controls were 76.6% and 76.8%, respectively, for the E1 allele (wild type) and 23.4% and 23.2%, respectively, for the E2 allele (polymorphic) (odds ratio 1.01; P > .99). The investigated polymorphism of the IL-1beta gene was not correlated with IL-1beta serum levels. CONCLUSIONS: We did not find an association between the IL1B exon 5 polymorphism, endometriosis, or increased serum IL-1beta levels.     

Low serum and peritoneal fluid concentration of interferon-gamma-induced protein-10 (CXCL10) in women with endometriosis

OBJECTIVE: To evaluate serum and peritoneal fluid concentrations of interferon-gamma-inducible protein-10 (CXCL10), a chemokine involved in local immune function, in women with endometriosis. DESIGN: Prospective study. SETTING: Division of Obstetrics and Gynecology, University of Siena. PATIENT(S): A total of 147 women were divided in two groups: women with (n = 77) and without (n = 70) endometriosis. INTERVENTION(S): Serum and peritoneal fluid were collected from all patients undergoing laparoscopy. MAIN OUTCOME MEASURE(S): CXCL10 concentrations were measured by a specific ELISA. RESULT(S): Serum CXCL10 concentrations in women with endometriosis were significantly lower than in those without endometriosis. No statistically significant difference between women with early endometriosis and those with advanced endometriosis was found. CXCL10 concentrations in peritoneal fluid of women with advanced endometriosis were significantly lower than in that of women with an early stage of, or without, endometriosis. CONCLUSION(S): The decreased concentrations of CXCL10 in serum and peritoneal fluid of women with endometriosis indicate an impaired immune activity in women with endometriosis.     

Concentrations of interleukin (IL)-1alpha, IL-1 soluble receptor type II (IL-1 sRII) and IL-1 receptor antagonist (IL-1 Ra) in the peritoneal fluid and serum of infertile women with endometriosis

OBJECTIVE: Endometriosis is an immune system-related gynaecological disease, characterised by an increase in number and activation of peritoneal macrophages. One of macrophage-derived factors is interleukin (IL)-1. The effects of IL-1 are inhibited by IL-1 receptor type II (IL-1 RII), soluble forms of IL-1 RII (IL-1 sRII) and IL-1 receptor antagonist (IL-1 Ra). The aim of our work was to study the IL-1alpha, IL-1 sRII and IL-1 Ra levels in the peritoneal fluid (PF) and serum of women with endometriosis in relation to stage of disease. STUDY DESIGN: Concentrations of IL-1alpha, IL-1 sRII and IL-1 Ra were measured by ELISA assay in the PF and serum of 58 women; 43 with and 15 without endometriosis (control group). RESULTS: Elevated PF and serum IL-1alpha and IL-1 Ra levels in the women with endometriosis in comparison with the control group were observed. IL-1 sRII levels in PF and serum were higher in the controls than in the women with endometriosis. Concentrations of IL-1alpha and IL-1 sRII were higher in advanced endometriosis, but higher IL-1 Ra was observed in the early stage of the disease. CONCLUSION: Impairment of regulation IL-1 activity in the peritoneal fluid and serum of women with endometriosis may play an important role in the pathogenesis and development of the disease.     

Elevated soluble Fas ligand levels may suggest a role for apoptosis in women with endometriosis

OBJECTIVE: To evaluate soluble Fas ligand concentrations in serum and peritoneal fluid from women with endometriosis and from fertile controls without endometriosis, and to study levels of soluble Fas ligand in conditioned media of cultured endometrial stromal cells. DESIGN: Prospective, experimental trial. SETTING: Two academic IVF centers. PATIENT(S): Twenty-nine fertile women without endometriosis and 57 infertile women with endometriosis (32 with stage I or II disease and 25 with stage III or IV disease). MAIN OUTCOME MEASURE(S): Enzyme-linked immunosorbent assay was used to measure soluble Fas ligand concentrations in paired samples of serum and peritoneal fluid from women with and without endometriosis. Concentrations were also measured in conditioned media of cultured endometrial stromal cells at basal conditions and after stimulation with interleukin-8 (0.001-10 ng/mL) and tumor necrosis factor-alpha (1-10 ng/mL). RESULT(S): Compared with fertile controls and women with early-stage of endometriosis, women with moderate to severe endometriosis had elevated serum (87.2 +/- 6.4, 88.2 +/- 6.9, and 162.3 +/- 7.8 pg/mL, respectively) and peritoneal fluid (81.0 +/- 6.0, 80.5 +/- 6.8, and 166.2 +/- 10.3 pg/mL, respectively) concentrations of soluble Fas ligand. Serum levels of soluble Fas ligand positively correlated with levels in peritoneal fluid. Comparison of patients in the same menstrual cycle in each group revealed that increased levels of soluble Fas ligand in patients with advanced endometriosis were not attributable to the difference in cycle phases. Soluble Fas ligand was not detected in conditioned media of endometrial stromal cells under baseline conditions or after stimulation. CONCLUSION(S): Serum and peritoneal fluid of women with moderate to severe endometriosis contain elevated concentrations of soluble Fas ligand compared to women with minimal or mild endometriosis and women without endometriosis. These findings suggest a role for apoptotic dysregulation in the pathophysiology of endometriosis.     

Association between endometriosis and polymorphisms in tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), TRAIL receptor and osteoprotegerin genes and their serum levels

Purpose

To evaluate the relationship between endometriosis and polymorphisms in the genes encoding tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL), TRAIL receptor (DR) and osteoprotegerin (OPG) and their serum levels in Korean women.

Methods

A case?Ccontrol study was conducted with 138 women with endometriosis and 214 women without endometriosis in academic medical center. TRAIL c.49G>A, c.592A>G, c.615A>G, and c.662T>C; DR4 c.626G>C and c.1322A>G; DR5 c.95C>T, c.200C>T, and c.72T>G; OPG ?245T>G, c.9C>G, c.788A>C, and c.9938G>T polymorphisms were investigated and circulating levels of TRAIL and OPG were measured.

Results

The TRAIL c.49G>A, c.615A>G, and c.662T>C; the DR4 c.626G>C; the DR5 c.72T>G; the OPG c.788A>C and c.9938G>T polymorphisms were not observed. The genotype distributions and allele frequencies of single or combined polymorphisms of TRAIL, DR4, DR5, and OPG measured in women with endometriosis were not different from those in women without endometriosis, regardless of endometriosis stage. Serum TRAIL and OPG levels were significantly lower in women with endometriosis than in women without endometriosis, but these levels did not show differences between early and advanced endometriosis.

Conclusions

Endometriosis is associated with circulating TRAIL and OPG levels in Korean women but not with the TRAIL, DR, and OPG polymorphisms.     

水通道蛋白1在子宫内膜异位症患者血清和腹腔液中的表达及意义

目的:研究子宫内膜异位症(EMT)患者血清和腹腔液中水通道蛋白1(AQP1)及血管内皮生长因子(VEGF)表达水平的变化及其相关性,探讨其在子宫内膜异位症发病中的作用。方法:采用酶联免疫吸附方法(ELISA),测定36例EMT患者和22例对照组血清和腹腔液中AQP1和VEGF的含量,进行相关性分析。结果:EMT组血清和腹腔液中AQP1、VEGF水平均高于对照组,差异均有统计学意义(P<0.05)。AQP1、VEGF在EMT组血清和腹腔液的表达均有显著正相关性(r=0.776,P=0.000;r=0.771,P=0.000);AQP1、VEGF在对照组血清和腹腔液的表达则均无相关性(r=-0.026,P=0.910;r=-0.040,P=0.860)。结论:AQP1在EMT患者血清和腹腔液中过表达,可能推动异位内膜黏附和侵袭,AQP1与VEGF可能通过促血管生成,共同促进异位子宫内膜种植存活,是EMT形成的基础。     

Serum and peritoneal lavage fluid CA-125 levels in endometriosis

Serum and peritoneal lavage fluid CA-125 levels were assayed in 20 women with endometriosis and 20 control women at the time of laparoscopy. Serum levels of CA-125 were significantly higher in women with endometriosis. Peritoneal lavage fluid CA-125 levels were significantly higher than serum levels but showed no significant difference between control and endometriosis patients. Peritoneal lavage fluid CA-125 did not follow expected dilution curves when attempts were made to validate the assay. Serum CA-125 levels were a more sensitive indicator of endometriosis, than peritoneal lavage fluid CA-125 levels.     

Spontaneous and stimulated secretion of monocyte chemotactic protein-1 and macrophage migration inhibitory factor by peritoneal macrophages in women with and without endometriosis

OBJECTIVE: To assess spontaneous and stimulated secretion of monocyte chemotactic protein-1 (MCP-1) and macrophage migration inhibitory factor (MIF) by peritoneal macrophages in women with and without endometriosis. DESIGN: Macrophages were isolated from the peritoneal fluid and cultured for different periods of time (6, 20, and 44 hours) without any stimulation to determine spontaneous secretion of MCP-1 and MIF. Macrophages were also exposed to 1 microg/mL lipopolysaccharide for 6 hours to evaluate the stimulated secretion of these cytokines. SETTING: Gynecology clinic and human reproduction research laboratory. PATIENT(S): Twelve fertile women and 11 women with endometriosis. INTERVENTION(S): Peritoneal fluid obtained at laparoscopy. MAIN OUTCOME MEASURE(S): Monocyte chemotactic protein-1 and MIF concentrations in the culture medium using ELISA. RESULT(S): Peritoneal macrophages of women with endometriosis demonstrated an increased capacity to secrete MCP-1 either spontaneously or after stimulation with lipopolysaccharide. They also showed a marked tendency for an increased secretion of MIF, but no statistically significant difference was found. CONCLUSION(S): Monocyte chemotactic protein-1 and MIF production by peritoneal macrophages may contribute to paracrine and autocrine activation and to macrophage accumulation in the peritoneal cavity of women with endometriosis. These mechanisms may exacerbate peritoneal inflammation and favor the growth of endometrial implants.     

T helper (Th)1, Th2, and Th17 interleukin pathways in infertile patients with minimal/mild endometriosis

In the present study, interleukin (IL)-10, IL-12, IL-17, and IL-23 levels were measured in serum and peritoneal fluid of women with minimal or mild endometriosis and compared with levels in controls without endometriosis. Higher IL-23 levels were encountered in the peritoneal fluid of women with endometriosis, suggesting a possible role of this cytokine in these women's infertility.     

Genetic polymorphism in the fibrinolytic system and endometriosis

OBJECTIVE: Although most women experience retrograde menses during their reproductive life, endometriosis develops only in a small percentage. We hypothesized that persistence of a fibrin matrix in peritoneal pockets, as a result of hypofibrinolysis, could allow menstrually deposited endometrial fragments to initiate endometriosis. Fibrinolysis is modulated by several factors, and polymorphisms in the plasminogen activator inhibitor-1 (PAI-1) gene are considered to be one of the important determinants. The objective of this study was to evaluate PAI-1 genotypes in a group of women with or without endometriosis. METHODS: In 118 women (75 with laparoscopically confirmed endometriosis and 43 controls), genomic DNA was extracted from blood and the PAI-1 promoter genotype was determined by polymerase chain reaction amplification of DNA using specific primers for the 4G or 5G allele followed by gel electrophoresis. A portion of the polymerase chain reaction product was purified and sequenced to confirm the gel electrophoresis results. RESULTS: Endometriosis was more likely in patients with 4G/5G (odds ratio 38; 95% confidence interval [CI] 6-229) or 4G/4G (odds ratio 441; 95% CI 53-3,694) compared with 5G/5G PAI-1 genotype. Fifty-two of 75 women with endometriosis (69 %, 95% CI 58-79%) had the 4G/4G genotype compared with only 5 of 43 (12%; 95% CI 4-25%) controls. In contrast, the 5G/5G genotype associated with normal fibrinolysis was found in 2 of 75 (3%; 95% CI 0-9%) women with endometriosis compared with 24 of 43 (56%; 95% CI 40-71%) controls. CONCLUSION: Hypofibrinolysis, associated with the 4G allele of the PAI-1 gene, was found significantly more often in women with endometriosis compared with controls. Persistence of fibrin matrix could support the initiation of endometriotic lesions in the peritoneal cavity, explaining why some women with retrograde menstruation develop endometriosis while others do not. LEVEL OF EVIDENCE: II-2.     

Association between human alpha 2-Heremans Schmidt glycoprotein (AHSG) polymorphism and endometriosis in Korean women

OBJECTIVE: To evaluate the relationship between the alpha 2-Heremans Schmidt glycoprotein (AHSG) gene polymorphism and endometriosis. DESIGN: Case-control study. SETTING: Department of Obstetrics and Gynecology, Seoul National University Hospital, Korea. PATIENT(S): Seventy-nine women with endometriosis and 105 women without endometriosis. INTERVENTION(S): Determination of AHSG gene polymorphism. MAIN OUTCOME MEASURE(S): Prevalence of AHSG genotypes or alleles. RESULT(S): The allele frequencies of AHSG 1 and AHSG 2 were found to be 0.69 and 0.31, respectively. The proportion of noncarriers of the AHSG 2 allele was significantly higher in women with endometriosis than in women without (55.7% vs. 39.0%). Women not carrying the AHSG 2 allele were found to have twice the risk of endometriosis than those carrying at least one copy of this allele. No significant difference was noted in the distribution of the AHSG alleles or AHSG genotypes between early stage endometriosis and late stage endometriosis. CONCLUSION(S): Endometriosis is associated with the AHSG gene polymorphism in Korean women.     

CA 125 in serum, peritoneal fluid, active lesions, and endometrium of patients with endometriosis

Ca 125 levels in serum and peritoneal fluid were measured in 39 patients with endometriosis and 18 patients with normal pelvic anatomy at laparoscopy, and the presence of this antigen in endometriotic tissue and endometrial mucosa was also investigated. Serum CA 125 concentrations were elevated in patients with Stage III or IV endometriosis compared with control subjects (32.9 +/- 11.2 versus 16.4 +/- 8.9 U/ml, means +/- SD; p less than 0.001). CA 125 values were greater than 35 U/ml in 36.8% of women with Stage III or IV endometriosis and in none of the control subjects. No significant differences in CA 125 levels in peritoneal fluid were found between patients with endometriosis and control subjects. The immunohistochemical studies found CA 125 in 10% of the endometriotic lesions and 37.5% of the endometrial samples of patients with endometriosis and in 33.3% of the endometrial samples of control subjects.     

Antizona and antisperm antibodies in women with endometriosis and/or infertility

OBJECTIVE: To measure the levels of antigamete antibodies in serum and peritoneal fluid of women with endometriosis and/or infertility. DESIGN: Antibody activity against human sperm and porcine oocytes was analyzed in selected subgroups of women. SETTING: Clinic of reproduction. PATIENT(S): Women with endometriosis and/or infertility. INTERVENTION(S): No treatment was implemented before peritoneal fluid and blood sample collection. MAIN OUTCOME MEASURE(S): Quantitative ELISA. RESULT(S): Four groups of women (n = 98) were analyzed for the presence of antizona and antisperm antibodies: infertile with endometriosis (n = 30), idiopathic infertility (n = 28), fertile with endometriosis (n = 20), and healthy fertile controls (n = 20). Antibodies were analyzed simultaneously in serum and peritoneal fluid. No statistically significant differences in antibody levels were detected in serum samples among the analyzed groups. The median values for antizona and antisperm antibodies in peritoneal fluid were significantly higher in women with idiopathic infertility than in the control group. In women with unexplained infertility, a high degree of correlation (Spearman) was found between the presence of antizona antibodies in peritoneal fluid and serum (r = 0.579). A positive predictive value of 80% was calculated for the presence of antizona antibodies (>5 ng/oocyte) in the peritoneal fluid of patients with infertility. CONCLUSION(S): Antizona antibodies locally produced in the peritoneal fluid have diagnostic value for infertility status; however, they cannot be treated as a marker or prognostic factor for minimal endometriosis and/or its treatment.     

AHSG和IL-6基因多态性与Ⅳ期卵巢子宫内膜异位囊肿的相关性研究

目的:探讨α2-HS-糖蛋白(AHSG)基因和白细胞介素-6(IL-6)基因调控区-174位点多态性与中国河北省育龄妇女Ⅳ期卵巢子宫内膜异位囊肿遗传易感性的关系。方法:采用病例对照研究的方法,以50例Ⅳ期卵巢子宫内膜异位囊肿患者(评分>40)及62例非子宫内膜异位症对照者的外周血白细胞为样本,利用聚合酶链反应(PCR)技术分析AHSG基因和IL-6基因调控区-174位点基因的多态性。结果:Ⅳ期卵巢子宫内膜异位囊肿组中AHSG基因的3种基因型AHSG1*1、AHSG1*2、AHSG2*2各占44%、46%、10%,对照组则分别为71%、25.8%、3.2%。将突变型与杂合型合并与野生型比较,两组间差异有统计学意义(χ2=8.317,P=0.004),其OR为3.111(95%CI为1.422~6.805),携带变异基因的患病风险率高。AHSG1、AHSG2等位基因频率总体分布差异有统计学意义(χ2=8.723,P=0.003),携带AHSG2等位基因的患病危险度高,OR为2.561,(95%CI为1.358~4.831)。在50例试验组与62例对照组中均未发现IL-6突变基因。结论:AHSG2基因可使患Ⅳ期卵巢子宫内膜异位囊肿的遗传易感性升高,未发现Ⅳ期卵巢子宫内膜异位囊肿与IL-6基因调控区-174位点突变有相关性。     

Cyclin D1 polymorphism and the risk of endometrial cancer

OBJECTIVES: The common G to A single nucleotide polymorphism (G870A) in the splice donor region of exon 4 enhances alternate splicing, and produces a longer half-life cyclin D1 (CCND1). This study was aimed at investigating the possible association between the G870A polymorphism in CCND1 and the risk of endometrial cancer. METHODS: We assessed the association between the CCND1 G870A polymorphism and the risk of endometrial cancer in a hospital-based case-control study among 231 Korean women (77 cases; 154 matched controls). Controls were matched to cases with respect to age, menopausal status, and hormone therapy status. RESULT: The allele frequencies of the case subjects (A, 0.45; G, 0.55) were significantly different from those of control subjects (A, 0.58; G, 0.42) (P = 0.008). All case and control subjects were in Hardy-Weinberg equilibrium. The AA genotype was associated with a significantly elevated odds ratio (OR) of 3.18 [95% confidence interval (CI) 1.38-7.37, P = 0.007], and the AG genotype was associated with an OR of 1.38 (95% CI 0.65-2.89). When we combined the GG and AG genotypes as a reference genotype, we found that the OR for the AA genotype was 2.53 (95% CI 1.34-4.80, P = 0.004), supporting a recessive model for the A allele. Conditional logistic regression adjusted for various risk factors of endometrial cancer revealed positive associations between the AA genotype and an increased risk of endometrial cancer (OR 3.16, 95% CI 1.18-8.43, P = 0.022). However, no significant difference in endometrial cancer stage or grade was observed between the CCND1 genotypes. CONCLUSION: Our data suggest that the CCND1 polymorphism is associated with an increased risk of endometrial cancer. To validate this association, a large-scale population-based study is needed.     

Serum and peritoneal fluid levels of interleukin-6 and interleukin-37 as biomarkers for endometriosis

This study aimed to compare the levels of interleukin (IL)-1β, IL-6, IL-10, and IL-37 in the serum and peritoneal fluid of women with and without endometriosis. In addition, it aimed to determine the diagnostic values of the cytokines with significantly different concentrations. The levels of IL-1β, IL-6, IL-10, and IL-37 in the serum and peritoneal fluid samples of 40 women with endometriosis and 32 women without endometriosis were measured using an enzyme-linked immunosorbent assay. The serum and peritoneal fluid levels of IL-1β and IL-10 were not statistically significantly different between the endometriosis and control groups. The IL-6 and IL-37 levels in the serum and peritoneal fluid were higher in the endometriosis group than in the control group, and they were correlated with the stage of endometriosis. The AUC for the IL-37 was 0.897 for the serum and 0.934 for the peritoneal fluid, while the AUC for the IL-6 was 0.905 for the serum and 0.952 for the peritoneal fluid. Our results suggest that the serum and peritoneal fluid IL-6 and IL-37 levels were significantly increased in the endometriosis patients, indicating that these cytokines may serve as biomarkers for the diagnosis of endometriosis.     

Use of serum-soluble intercellular adhesion molecule-1 as a new marker of endometriosis

OBJECTIVE: To investigate the hypothesis that soluble intercellular adhesions molecule-1 (sICAM-1) may be used as a new serum marker of endometriosis. DESIGN: Prospective cohort study. SETTING: An academic department specializing in gynecologic laparoscopy. PATIENT(S): Consecutive series of 120 women of reproductive age who underwent laparoscopy for benign gynecologic conditions. INTERVENTION(S): Data were collected on baseline clinical characteristics, and surgical and histologic diagnosis. MAIN OUTCOME MEASURE(S): Serum concentration of both CA125 and sICAM-1. RESULT(S): Endometriosis was documented in 71 women (stage I to II in 24 cases and stage III to IV in 47 cases). Serum levels of sICAM-1 were only slightly but not significantly increased in women with endometriosis compared with women without the disease. However, serum concentration of sICAM-1 in the 21 women who were found to have deep peritoneal endometriosis was significantly enhanced when compared with both women without the disease and those with other forms of endometriosis. The sensitivity and specificity of sICAM-1 in detecting deep peritoneal endometriosis were 0.19 and 0.97, respectively; whereas those of CA125 were 0.14 and 0.92, respectively. When both parameters were used concomitantly, the sensitivity and specificity were 0.28 and 0.92, respectively. CONCLUSION(S): Although the present study tends to support a role of sICAM-1 in the development of endometriosis, serum concentrations of this molecule do not seem to be an effective indicator for the diagnosis of either the early or advanced stage of endometriosis. However, an integrated clinical and laboratory approach using both CA125 and sICAM-1 may be helpful in specifically identifying women with deep peritoneal endometriosis.