Serum levels of interleukin-6, interleukin-1β and human chorionic gonadotropin in pre-eclamptic and normal pregnancy

Studies in placentas from the first trimester and in vitro models indicate that interleukin (IL)-1β and IL-6 induce the release of human chorionic gonadotropin (hCG). During pre-eclampsia there is an increase of pro-inflammatory cytokines; however, its relationship with hCG levels during the third trimester of pregnancy has not been determined. The aim of the present study was to evaluate the relationship between blood levels of IL-6, IL-1β and hCG in normal pregnancy and pre-eclampsia. Blood samples during the third trimester of pregnancy from women with severe pre-eclampsia (n = 20) or normal pregnancy (n = 20) were assayed for hCG by immunoassay, IL-6 and IL-1β by enzyme-linked immunosorbent assay. Serum level of IL-6 was significantly higher in pre-eclamptic than in normal women (16.5 ± 2.1 vs. 4.9 ± 1.1 pg/ml); however, IL-1β was similar in both groups. Although hCG was higher in pre-eclampsia than normal pregnancy, the difference was not statistically significant. Furthermore, IL-1β in normal pregnancy was correlated negatively with hCG (r = ?0.69, p < 0.001). In conclusion, serum levels of IL-6 were increased in pre-eclampsia but were not correlated with hCG or IL-1β; however, in normal pregnancy there was a negative correlation between IL-1β and hCG. The interaction between IL-1β and hCG at the third trimester needs to be investigated.     

Do alterations in follicular fluid proteases contribute to human infertility?

Purpose

Cathepsin L and ADAMTS-1 are known to play critical roles in follicular rupture, ovulation, and fertility in mice. Similar studies in humans are limited; however, both are known to increase during the periovulatory period. No studies have examined either protease in the follicular fluid of women with unexplained infertility or infertility related to advanced maternal age (AMA). We sought to determine if alterations in cathepsin L and/or ADAMTS-1 existed in these infertile populations.

Methods

Patients undergoing in vitro fertilization (IVF) for unexplained infertility or AMA-related infertility were prospectively recruited for the study; patients with tubal or male factor infertility were recruited as controls. Follicular fluid was collected to determine gene expression (via quantitative polymerase chain reaction), enzyme concentrations (via enzyme-linked immunosorbent assays), and enzymatic activities (via fluorogenic enzyme cleavage assay or Western blot analysis) of cathepsin L and ADAMTS-1.

Results

The analysis included a total of 42 patients (14 per group). We found no statistically significant difference in gene expression, enzyme concentration, or enzymatic activity of cathepsin L or ADAMTS-1 in unexplained infertility or AMA-related infertility as compared to controls. We also found no statistically significant difference in expression or concentration with advancing age.

Conclusions

Cathepsin L and ADAMTS-1 are not altered in women with unexplained infertility or AMA-related infertility undergoing IVF, and they do not decline with advancing age. It is possible that differences exist in natural cycles, contributing to infertility; however, our findings do not support a role for protease alterations as a common cause of infertility.     

Increased levels of interleukin-10 and transforming growth factor-β in the plasma and ascitic fluid of patients with advanced ovarian cancer

Objectives To assess expression of the immunosuppressive cytokines IL-10 and TGF-β in the ascitic fluid and plasma of advanced ovarian cancer patients.Setting The Department of Obstetrics and Gynaecology at the University of Arkansas for Medical Sciences.Population Twenty-eight women diagnosed with advanced ovarian cancer and ten normal female controls.Methods Plasma and ascitic samples were collected at the time of surgery and analysed for the presence of IL-10 and TGF-β using a sensitive enzyme-linked immunosorbent assay.Results Elevated levels of IL-10 were detected in the plasma [mean (SD) = 12 (5) pg/mL; range 8 to 23 pg/mL] and in the peritoneal fluid [mean (SD) = 165 (137) pg/mL; range 50 to 556 pg/mL] of ovarian cancer patients, while no detectable IL-10 was found in any of the normal control plasma samples tested. Similarly, plasma levels of TGF-β in ovarian cancer patients were significantly higher [mean (SD) = 1506 (246) pg/mL; range 1020 to 2070 pg/mL] compared with controls [mean (SD) = 937 (187) pg/mL; range 770 to 1140 pg/mL](P < 0.001). Surprisingly, however, although elevated TGF-β levels were also detected in the peritoneal fluid of all ovarian cancer patients [mean (SD) = 407 (158) pg/mL; range 140 to 770 pg/mL], these levels were significantly lower than those seen in matched plasma samples (P < 0.001).Conclusions Local and systemic secretion of immunosuppressive cytokines may play an important role in the impaired anti-tumour immune function commonly observed in advanced ovarian cancer. However, the observation that plasma levels of TGF-β are significantly higher than those detected in the ascitic fluid raises the possibility that cells other than tumour cells are responsible for TGF-β release in the bloodstream of these patients.     

Do follicular fluid gelatinase levels affect fertilization rates and oocyte quality?

Background

Matrix metalloproteinase-2 and -9, known as gelatinases, are considered to be essential for tissue remodelling during the reproductive process. However, their role in reproduction is unclear.

Aims

In the present study, we aimed to investigate the relationship between follicular fluid gelatinase levels and oocyte quality and fertilization, and to compare the activities of gelatinase levels with different drug stimulation protocols.

Methods

We evaluated 60 women with unexplained infertility who underwent in vitro fertilization (IVF) treatment. Thirty patients underwent a gonadotropin-releasing hormone (GnRH) agonist protocol and 30 underwent a GnRH antagonist protocol. Follicular fluid was obtained during oocyte retrieval. Oocyte quality was determined using light microscopy, and oocytes were considered to be fertilized when two pronuclei were present. Gelatinase activities were measured using commercial enzyme-linked immunosorbent assay kits. The study was partially supported by the Scientific Research Unit of Suleyman Demirel University (Protocol Number: 3620-TU1-13), and all procedures were conducted with the approval of the Suleyman Demirel University Local Ethics Board. Statistical analyses of the data were performed using the independent t test, Fisher exact test, Mann–Whitney U test, one-way ANOVA, and posthoc least significant difference.

Results

Follicular fluid gelatinase levels were significantly higher for agonist drug administration (p = 0.001), and a positive correlation was observed between matrix metalloproteinase-9 levels and oocyte grade (p = 0.01). Moreover, a positive relationship between matrix metalloproteinase-9 levels and fertilization was observed (p = 0.02).

Conclusion

Follicular fluid gelatinase activities, particularly MMP-9 activity, might be a predictor of oocyte quality and IVF success.     

Maternal and umbilical serum levels of interleukin-6, interleukin-8, and tumor necrosis factor-α in normal pregnancies and in pregnancies complicated by preeclampsia

Objectives.?The aim of this study was to investigate the relationship of maternal and umbilical cord interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α) serum levels with the existence and severity of preeclampsia. A particular objective was the comparison of normal umbilical serum levels to preeclamptic values.

Materials and Methods.?The study group consisted of 24 patients with third trimester singleton pregnancies complicated by preeclampsia (15 severe and 9 mild preeclampsia). The gestational age-matched 19 healthy pregnant women were compared by study group. Maternal and umbilical serum IL-6, IL-8, and TNF-α were calculated by using enzyme-linked immunosorbent assay.

Results.?Significantly increased maternal and umbilical serum levels of IL-6, IL-8, and TNF-α were found in preeclamptic patient group in comparison with the control group. Maternal serum IL-8 and TNF-α concentration were significantly higher in patients with severe preeclampsia than in mild preeclampsia. Increased umbilical serum levels of IL-6 and IL-8 were found in severe preeclampsia than in mild preeclampsia. There were significantly higher levels of maternal serum IL-8 and TNF-α in patients with preeclampsia with IUGR than in patients with preeclampsia with normal fetal growth.

Conclusion.?Our findings suggest that increased concentrations of IL-6, IL-8, and TNF-α in the maternal and umbilical serum play a significant role in pathogenesis of preeclampsia. Alterations in maternal and umbilical serum levels of IL-6, IL-8, and TNF-α may also play role in preeclampsia complicated by intrauterine growth retardation. These associations may offer insight into the etiology and pathogenesis of preeclampsia.     

Evidence that granulosa cells inhibit interleukin-1α and interleukin-2 production from follicular lymphomonocytes

Objective Relationships between immune and endocrine systems seem to occur in ovarian follicular fluids (FF). Lymphomonocytes have been found in preovulatory follicles and their specific products, cytokines [interleukin-1 (IL1), IL2], were demonstrated to inhibit steroidogenesis. Ovarian steroids, in turn, reduce the cytokine production from immune-competent cells. In the present study we evaluated whether lymphomonocytes are present in FF, and if both their subset distribution and their IL1 and IL2 secretions, after activation with phytohemagglutinin (PHA), are similar to those of peripheral blood. Interferences of IL1 and IL2 production by FF lymphomonocytes caused by isolated granulosa cells were also evaluated.Participants The study was performed on 86 FFs obtained from follicles containing mature oocytes that were aspirated at the time of ovum pickup from 27 women undergoing controlled ovarian hyperstimulation with exogenous gonadotropins for an in vitro fertilization (IVF) program [IVF—embryo transfer (IVF-ET) or gamete intrafallopian transfer (GIFT)].Results Lymphocytes were found in FF. The distribution of CD8+ and CD3+ lymphocyte subsets is equal to that in peripheral blood, but the percentage of CD11b+, CD16+, and CD4+ cells (its trend) is higher in FF than in peripheral blood. The amount of IL2 and IL1 deriving from PHA-activated FF lymphomonocytes is similar to that of peripheral blood PHA-activated lymphomonocytes. Granulosa cells significantly blunt IL2 and IL1 production by FF lymphomonocytes.Conclusions These results suggest that preovulation, a migration of lymphomonocytes from the peripheral compartment to the follicle occurs. However, unfavorable effects of IL2 and IL1, cytotoxic and antisteroidogenetic activities, are counteracted by the products of granulosa cells. The higher amounts of CD11b+, CD16+, and CD4+ in FF suggest that they could be involved in other immune processes.     

Apoptosis, reactive oxygen species and follicular anti-Müllerian hormone in natural versus stimulated cycles

The aim of this study was to compare granulosa cell apoptosis, reactive oxygen species (ROS) and anti-Müllerian hormone (AMH) production in natural and stimulated cycles, and to determine their effect on IVF outcome. Of 60 women with tubal factor infertility, 30 underwent natural cycle IVF and 30 underwent ovarian stimulation with gonadotrophin-releasing hormone antagonist and gonadotrophin. Apoptosis and ROS production was measured in isolated granulosa cells from each aspirated follicle using flow cytometry. Follicular AMH concentration was measured using enzyme immunoassay. A total of 188 follicles were analysed. A lower incidence of apoptotic granulosa cells and higher mean follicular AMH concentration was observed in the natural cycle than in the ovarian stimulation group (20.2 +/- 15.6% versus 35.6 +/- 27.7%, P = 0.007; 6.9 +/- 7.1 ng/ml versus 2.5 +/- 1.7 ng/ml, P < 0.001). Ovarian stimulation increases the incidence of apoptotic granulosa cells, but has no effect on their ROS production. Ovarian stimulation reduces follicular AMH concentrations, presumably due to a negative effect of exogenous FSH on AMH production. This study was unable to explain the low pregnancy rate in natural cycle IVF using the data on apoptosis, ROS and AMH production in granulosa cells.     

Sex hormone levels, genetic androgen receptor polymorphism, and anxiety in ≥50-year-old males

IntroductionWhile associations between somatic changes and sex hormone levels in aging men have been explored in many studies, the association of testosterone and estradiol with psychic symptoms other than depression and the role of the genetically determined CAG repeat (CAGn) polymorphism of the androgen receptor (AR) have received much less attention.AimThe purpose of this article is to investigate the associations between general anxiety, phobic anxiety and panic with sex hormone levels and the genetic androgen receptor polymorphism in aging males.MethodsThis cross-sectional study of males aged ≥50 years included 120 consecutive patients of the Department of Psychosomatics and Psychotherapy, 76 consecutive patients of the Andrology Clinic, and 100 participants from the general population; all of them completed the Brief Symptom Inventory (BSI), the Aging Males' Symptoms (AMS) Scale, and the Patient Health Questionnaire (PHQ-9). Morning blood samples were analyzed for total and free testosterone, estradiol, sex hormone-binding globulin (SHBG), and the CAGn AR polymorphism. Psychosomatic patients also underwent psychiatric assessment.Main Outcome MeasuresScores on the Anxiety subscales of the BSI and PHQ, Anxiety disorders according to International Classification of Diseases, 10th revision (ICD-10).ResultsThe two clinical samples had significantly longer CAGn of the AR and higher anxiety levels compared to the population sample. Anxiety scores were positively correlated with CAGn in psychosomatic patients and in andrological patients, in the latter also with estradiol levels, while the population sample showed no significant correlations between anxiety scores, CAGn and sex hormones. Anxiety cases according to BSI, PHQ, and ICD-10 had significantly longer CAGn of the AR when compared to the other participants, but there were no significant differences in testosterone or free testosterone levels.ConclusionsOur results indicate that genetically determined long CAGn of the AR is an independent risk factor for higher anxiety, panic and phobic anxiety levels. Schneider G, Nienhaus K, Gromoll J, Heuft G, Nieschlag E, and Zitzmann M. Sex hormone levels, genetic androgen receptor polymorphism and anxiety in ≥50-year-old males. J Sex Med **;**:**–**.     

Vascular endothelial growth factor in the plasma,follicular fluid and granulosa cells of women with endometriosis submitted to in vitro fertilization – a pilot study

Vascular endothelial growth factor (VEGF), a potent angiogenic factor that is altered in endometriosis, supports the immunological mechanism involved in this disease. The aim of the present study was to assess VEGF concentration in the plasma, follicular fluid (FF) and culture medium (CM) of granulosa cells from patients with endometriosis submitted to in vitro fertilization (IVF). A case–control study was conducted on 14 patients with endometriosis and 14 women without endometriosis submitted to IVF. Peripheral blood samples were collected before and after administration of human chorionic gonadotropin (hCG), in addition to FF and CM samples. Plasma VEGF levels increased after hCG administration in women with endometriosis and in controls, but were significant only in controls. VEGF levels were lower in FF but were significantly increased in the CM of patients with endometriosis. There was no correlation between VEGF and age, response to ovarian stimulation, oocyte or embryo quality, and pregnancy result. The increase of VEGF levels after hCG in both groups demonstrated a positive effect of this hormone on VEGF. VEGF in the FF and CM presented opposite results in endometriosis, suggesting that granulosa cells may show a different behavior in vivo and in vitro.     

The role of interleukin-1β in human trophoblast motility

The pleiotropic cytokine interleukin-1β (IL-1β) can promote physiological cell migration, as well as cancer cell invasion and metastasis. Its role in human trophoblast invasion, however, has not been satisfactorily answered since direct, indirect as well as no effects on trophoblast motility have been published. Therefore, the role of IL-1β has been re-evaluated by exclusively using human primary trophoblast model systems. Immunofluorescence of first trimester placentae indicated IL-1 receptor 1 (IL-1R1) protein expression in first trimester villous cytotrophoblasts (vCTB) and extravillous trophoblasts (EVT). The latter expressed higher mRNA levels of the receptor as shown by comparative gene chip data of vCTB and EVT. Similarly, Western blot analyses and immunofluorescence revealed a time- and differentiation-dependent increase of IL-1R1 in primary EVT seeded on fibronectin. IL-1β dose-dependently elevated migration of isolated first trimester EVT through fibronectin-coated transwells, which was inhibited in the presence of IL-1R antagonist (IL-1Ra), whereas proliferation of these cells was not affected. Similarly, the interleukin did not alter proliferation of vCTB and cell column trophoblasts in floating villi of early pregnancy, but promoted migration in villous explant cultures seeded on collagen I. Western blot analyses of supernatants of primary EVT and first trimester villous explant cultures revealed IL-1β induced secretion of urokinase plasminogen activator (uPA), plasminogen activator inhibitor (PAI)-1 and PAI-2, which was diminished upon combined IL-1β/IL-1Ra treatment. In conclusion, these data suggest that IL-1β directly promotes trophoblast motility of first trimester EVT involving the uPA/PAI system.     

Transforming growth factor-β1 in the human endometrium

Transforming growth factor-β ₁ (TGF-β₁) is a polypeptide involved in a variety of important physiological and pathophysiological processes such as the implantation of the embryo into the endometrium. Many factors seem to be related to this event. TGF-β₁ is involved in many mechanisms both in endometrial and in embryonic tissues: it induces proliferation and differentiation, it regulates proteolytic activity and it modulates the maternal immune response. Tins study evaluated the presence of TGF-β₁ in the endometrium during normal menstrual cycles and in the uterine fluids during induction of ovulation in the framework of an in vitro fertilization program. Immuno-histochemistry was used to identify TGF-β₁ in the endometrium and immunodot-blot to auantitate TGF-β₁ in the uterine cavity fluid. The study shows that TGF-β₁ is present in the endometrial tissue and its secretion is modulated during the menstrual cycle, as demonstrated immunohistochemically; its production seems to be controlled by ovarian steroids. In conclusion, TGF-β₁ influences the growth and differentiation of the embryo, as well as the activation of embryonic proteolytic enzymes, and it modulates the maternal–embryonic immune response. Its variability in the uterine cavity is demonstrated in this study, and the underexpression of TGF-β₁ in the uterine cavity might be responsible for failed implantation.     

Interleukin-1β regulates vascular endothelial growth factor and soluble fms-like tyrosine kinase-1 secretion by human oviductal epithelial cells and stromal fibroblasts

The aim of the present study was to evaluate the effects of interleukin (IL)-1β on the production of vascular endothelial growth factor (VEGF) and soluble fms-like tyrosine kinase-1 (sFlt-1) in the human fallopian tube. Human oviductal epithelial cells (OECs) and oviductal stromal fibroblasts (OSFs) were isolated from ten premenopausal patients. The secretion of VEGF and sFlt-1 by cultured OECs and OSFs in response to IL-1β and IL-1 receptor antagonist (IL-1RA) was measured using an enzyme-linked immunosorbent assay. The secretion of VEGF and sFlt-1 was detected in cultured OECs and OSFs under untreated conditions; secretion of these angiogenic modulators was significantly stimulated with IL-1β administration in these cells. IL-1β-induced production of VEGF and sFlt-1 by these cells was significantly inhibited by the addition of IL-1RA. The present findings suggest that IL-1β in the local environment may stimulate oviductal vascular permeability by inducing the production of VEGF by oviductal cells via both autocrine and paracrine mechanisms. Simultaneous upregulation of sFlt-1 secretion by these cells after IL-1β stimulation may prevent an excessive upregulation of vascular permeability. The modulation of the ratio of VEGF and sFlt-1 in the fallopian tube may contribute to the normal and pathological processes of oviductal fluid secretion by regulating oviductal vascular permeability during the menstrual cycle and during the peri-implantation period.