Endometrial Cytokine Expression Research Articles

O-056 Can we tailor ART to endometriosis patients?

Abstract ART remains an effective treatment for endometriosis-associated infertility, although there is evidence that pregnancy rates are diminished in women with endometriosis compared with other etiologies of infertility. In this lecture, the literature relating to endometriosis-associated infertility will be evaluated and recommendations will be made on the management of patients both pre-ART and during ART in order to improve ART outcomes in women with endometriosis. Surgery as an adjuvant to ART appears to have a favorable effect in patients with minimal to mild endometriosis prior to IVF. However, this evidence is based on only one study and more research is needed. As deep endometriosis (DE) is usually accompanied by advanced intra-abdominal disease resulting in distortion of the pelvic anatomy and tubal dysfunction, it is not surprising that IVF is considered as first line treatment. Observational data are available on the outcomes of DE surgery regarding conception rates in infertile patients with endometriosis suggesting that surgery may increase natural pregnancy rates as well as improve IVF outcomes. However, randomized trials comparing IVF to DE surgery are non-existent, as DE surgery is still mainly performed for pain and reduced quality of life rather than for treating infertility. In order to improve IVF success rates in endometriosis, various pre-ART treatments have been suggested. The oldest one relates to the use of gonadotropin releasing hormone (GnRH) agonist prior to IVF. Currently there is uncertainty as to whether long-term GnRH agonist therapy is beneficial when compared to standard IVF/ICSI in endometriosis. In addition, there is no evidence to support the use of oral GnRH antagonists and oral contraceptives as pre-treatment prior to ART, but results of ongoing studies will determine further. More recent studies support defects in endometrial receptivity as a cause of IVF failure. In recent years a new marker for endometrial receptivity in endometriosis emerged: BCL6, a biomarker for endometrial inflammation as it stimulates endometrial cytokine expression. Prospective cohort data provide a proof of concept that high BCL6 expression is associated with adverse IVF outcomes in women with endometriosis and that patients with high BCL6 expression may benefit from medical and surgical treatment prior to IVF. In order to improve IVF success rates in endometriosis, various pre-ART treatments have been suggested. The oldest one relates to the use of gonadotropin releasing hormone (GnRH) agonist prior to IVF. Contrary to previous findings, there is currently uncertainty as to whether long-term GnRH agonist therapy is beneficial when compared to standard IVF in endometriosis. In addition, there is also no evidence to support the use of oral GnRH antagonists and oral contraceptives as pre-treatment prior to ART in endometriosis, but results of ongoing studies will determine further. It has been hypothesized that applying local endometrial injury might induce a beneficial effect on endometrial receptivity prior to ART. However, scratching the endometrium as well as infusing fluids (Lipiodol and ExEm gel) into the uterine cavity (uterine bathing) in endometriosis patients prior to ART did not improve IVF success. During ART, a specific protocol for ART in women with endometriosis cannot be recommended. Both GnRH agonist and antagonist protocols can be used. For ovarian stimulation, a higher dose of gonadotropins may be required in stage III-IV endometriosis due to a poorer ovarian response. There is insufficient evidence for the routine use of prophylactic antibiotics at the time of oocyte retrieval in patients with endometriosis, but their use is recommended in patients with endometrioma. Considering fertilization, there appears no added benefit in using ICSI over IVF in endometriosis unless male-factor infertility is present. Finally, there is insufficient evidence to recommend any particular luteal support in women with endometriosis undergoing ART.

Endometrial cytokine expression from clinically suspected genital tuberculosis patients at tertiary care hospitals in Dhaka

ObjectiveThe objective of this study was to measure gamma interferon (IFN-γ) and tumor necrosis factor alpha (TNF-α) expression in endometrial tissue and/or aspirate from suspected genital tuberculosis patients with ectopic pregnancy and infertility in Bangladesh. MethodologyA total 78 women of clinically suspected genital tuberculosis patients were enrolled as study population. These patients underwent manual vaccum aspiration (MVA) procedure, and endometrial tissues and/or aspirates were collected. Ziehl -Neelsen staining (Z-N staining) and Lowen–Stein Jensen (L-J) culture were done to detect Mycobacterium. The study participants were categorized as genital tuberculosis positive cases, genital tuberculosis negative cases and presumptive for tuberculosis cases based on the case definition used in this study. TNF-α and IFN-γ were measured by ELISA. Statistical analysis was done using SPSS (version-22). ResultsOut of 78 participants, pro-inflammatory cytokines IFN-γ and TNF-α were significantly increased in TB positive patients than TB negative patients (p < 0.05). IFN-γ value of TB positive patients (41.26 ± 41.05) was higher than TB negative (22.94 ± 44.51) patients. TNF-α value (44.31 ± 64.22) of TB positive patients was higher than TB negative (15.86 ± 41.45) patients. IFN-γ and TNF-α value of presumptive for tuberculosis cases were not statistically significant. According to ROC analysis, cut off value for IFN-γ was 23.5 and for TNF-α was 10 with highest sensitivity and specificity of 66.7%, 89.3%, and 66.7% and 73.1% respectively. ConclusionIFN-γ and TNF-α were significantly higher in TB positive patients and it may act as a potential biomarker for diagnosis of genital tuberculosis.

Our current understanding of the pathophysiology of equine endometritis with an emphasis on breeding-induced endometritis

Equine endometritis is characterized by the expression of pro- and anti- inflammatory cytokines and an influx of polymorphonuclear neutrophils (PMNs) into the uterus. Mares resistant to persistent endometritis clear the inflammation within 24–36h after exposure to microorganisms or semen. These mares have a rapid increase of pro-inflammatory cytokines and an upregulation of inflammatory modulating cytokines within 6h after exposure to inflammatory challenge. In conjunction with effective uterine contractions, these events are believed to be responsible for the transient nature of the inflammation. In contrast, mares that are susceptible to persistent endometritis fail to clear the inflammation in a timely fashion. They have an imbalanced endometrial mRNA expression of pro- and anti-inflammatory cytokines, and have also been shown to suffer from an accumulation of intraluminal nitric oxide, which may be related to impaired myoelectrical activity and delayed uterine clearance. As a consequence, these mares establish a chronic inflammation, which interferes with the establishment of pregnancy. Recent studies on endometrial cytokine expression in resistant and susceptible mares have revealed information that suggest an underlying immunologic basis for susceptibility to persistent endometritis. However, the inflammatory pathways have yet not been fully studied, and a relationship between cytokine expression, nitric oxide, and myometrial contractions has not been established. In addition, seminal plasma has been shown to modulate breeding induced inflammation, but the molecular basis of the modulation is not understood. A holistic approach appears to be needed to better understand the characteristics of inflammatory pathways and ultimately the pathophysiology of the disease.

The effect of treatment with immune modulators on endometrial cytokine expression in mares susceptible to persistent breeding‐induced endometritis

Research has shown that 6 h after breeding is a critical time during the uterine innate immune response, and the failure to respond appropriately will result in persistent breeding-induced endometritis. This presents a potential opportunity to modulate the course of inflammation towards a timely resolution. To evaluate the effects of immune modulation on endometrial mRNA expression of inflammatory genes in susceptible mares 6 h after breeding. The hypothesis was that immune modulation alters endometrial cytokine expression in susceptible mares. A randomised controlled study to evaluate the effects of mycobacterial cell wall extract and dexamethasone on endometrial gene expression after insemination in 6 mares susceptible to persistent breeding-induced endometritis. Six susceptible mares were selected based on their uterine inflammatory response to insemination. Mares were inseminated during 3 oestrous cycles with 1 × 10(9) nonviable spermatozoa and 1) no additional treatment (control), or in combination with 2) dexamethasone (50 mg i.v.) at the time of insemination, or 3) with mycobacterial cell wall extract (1.5 ml i.v.) administered 24 h prior to insemination. Mares received one treatment per cycle in randomised order, and each mare served as her own control. Endometrial biopsies were collected 6 h after breeding, and quantitative polymerase chain reaction analysis for interleukin (IL)1β, IL6, interferon γ, IL10 and IL1RA was performed. Relative quantification values reported fold changes in mRNA expression from the control. Data were analysed using an ANOVA and significance was set at P<0.05. Expression of IL1β mRNA was lower after treatment with dexamethasone (P<0.001) and mycobacterial cell wall extract (P<0.05) when compared with control. No differences were detected in the mRNA expression of the other cytokines after any of the treatments. Treatment with immune modulators alters endometrial mRNA expression of IL1β after insemination. A better understanding of the mechanisms of immune modulation in the equine uterus can help to improve treatments for persistent breeding-induced endometritis.

Influence of dietary algal N-3 fatty acids on breeding induced inflammation and endometrial cytokine expression in mares bred with frozen semen

Influence of dietary algal N-3 fatty acids on breeding induced inflammation and endometrial cytokine expression in mares bred with frozen semen

Endometrial Cytokine Expression in Mares with Different Resistance to Persistent Breeding Induced Endometritis (PBIE) at Multiple Time Points after Insemination.

Endometrial Cytokine Expression in Mares with Different Resistance to Persistent Breeding Induced Endometritis (PBIE) at Multiple Time Points after Insemination.

Association between endometritis and endometrial cytokine expression in postpartum Holstein cows

The endometrium regulates the inflammatory response after infection by production and release of cytokines and chemokines. The objective was to compare gene expression of important pro-inflammatory (TNFα, IL-1β, IL-6) and anti-inflammatory (IL-10) cytokines, and the main neutrophil chemokine (IL-8), from calving to Week 7 after calving, in cows that developed endometritis and healthy control cows. Uterine biopsies were obtained at calving and at Weeks 1, 3, 5 and 7. Endometritis was evaluated at Week 5 by uterine lavage and cytology; cows with ≥ 10% neutrophils were considered to have endometritis. Real-time RT-PCR threshold values (Ct) were used to calculate the fold difference in gene expression, using the 2−ddCt method, normalized to GAPDH and calibrated to the average dCt for all cows at calving. Serum IL-8 concentrations were measured with ELISA. The analysis included 28 cows (11 had endometritis) for the PCR data and 44 cows (20 had endometritis) for ELISA. Expression of the TNFα gene in uterine tissue was decreased in cows with endometritis compared to control cows at calving (P = 0.09) and at Week 1 (P = 0.05). Iterleukin-1β gene expression tended to be decreased (P = 0.08) in cows with endometritis compared to control cows at Week 1, but tended to be increased (P ≤ 0.10) at Weeks 5 and 7. Cows with endometritis had increased (P < 0.05) IL-6 gene expression at calving and at Week 7 compared to control cows. Interleukin-8 gene expression was increased (P = 0.03) in endometritic cows compared to control cows at Week 7. Uterine disease was not significantly associated with IL-10 gene expression. A lower local level of expression of pro-inflammatory cytokines in the endometrium soon after calving might impair activation of inflammation and clearance of bacteria, and lead to development of endometritis.

Cyclic regulation of T-Bet and GATA-3 in human endometrium.

Endometrial cytokine expression is poorly understood. T-Bet and GATA-3 regulate cytokine expression in T-lymphocytes. Previous work has demonstrated expression of T-Bet in human endometrium. Changes in human endometrial T-Bet and GATA-3 mRNA and protein expression during the normal menstrual cycle were characterized. Human endometrium from each phase of the menstrual cycle underwent real-time reverse-transcriptase polymerase chain reaction and immunohistochemistry to examine expression and localization. T-Bet and GATA-3 mRNA were increased in the late secretory phase. Progesterone receptor (PR) mRNA was increased during the proliferative and early secretory phases. T-Bet and GATA-3 proteins localized cytoplasmically in the late secretory phase. PR protein displayed nuclear localization and maximal immunostaining during the early secretory phase. T-Bet and GATA-3 are expressed in endometrial epithelium cyclically during the menstrual cycle. T-Bet and GATA-3 are both upregulated during the late secretory phase and in the same cell types. The expression patterns of T-Bet and GATA-3 oppose PR, suggesting antagonistic function and/or regulation between PR and T-Bet/GATA-3.

Seminal plasma regulates endometrial cytokine expression, leukocyte recruitment and embryo development in the pig

In pigs, uterine exposure to the constituents of semen is known to increase litter size but the underlying physiological mechanisms remain undefined. Studies in rodents and humans implicate immune modulating moieties in seminal plasma as likely candidates, acting through enhancing the receptivity of the female tract. In this study, the acute and longer term effects of seminal plasma on cytokine expression and leukocyte abundance in the pig endometrium during early pregnancy have been characterised. The reproductive tracts of gonadotrophin-primed pre-pubertal gilts treated with intrauterine infusions of either pooled seminal plasma or phosphate-buffered saline (PBS) were retrieved at 34 h, or on day 5 and day 9 after treatment. Seminal plasma elicited an endometrial inflammatory infiltrate comprised of predominantly macrophages and major histocompatibility complex class II+-activated macrophages and dendritic cells. The abundance of these cells was greatest at the pre-ovulatory (34 h) time-point and their increase relative to PBS-treated tissues was maintained until day 9 after seminal plasma treatment. Seminal plasma induced the expression of the cytokines, granulocyte macrophage colony-stimulating factor, interleukin-6 and monocyte chemoattractant protein-1, and the eicosanoid-synthesising enzyme cyclo-oxygenase-2. Expression was maximal 34 h after treatment but altered expression patterns as a consequence of seminal plasma induction persisted through early pregnancy. These changes were accompanied by altered dynamics in pre-implantation embryo development with an increase in the number of embryos and in their viability after seminal plasma treatment. Together, these findings implicate factors in seminal plasma in programming the trajectory of uterine cytokine expression and leukocyte trafficking during early pregnancy and in regulating pre-implantation embryo development in the pig.

Seminal plasma induces mRNA expression of IL-1beta, IL-6 and LIF in endometrial epithelial cells in vitro.

The influence of seminal plasma on the mRNA expression of cytokines in human endometrial epithelial and stromal cells and the cytokine production of spermatozoa were investigated in vitro. Seminal plasma and spermatozoa were collected from healthy volunteers and were screened by enzyme-linked immunosorbent assay for cytokines. Epithelial and stromal cells from fertile women were cultured on matrigel or polystyrol and incubated with pooled seminal plasma or with transforming growth factor beta1 (TGF-beta1), interleukin 8 (IL-8) and vascular endothelial growth factor (VEGF), which were found to be significantly concentrated in seminal plasma. Endometrial cytokine expression was analysed by RNase protection assay and supported by RT-PCR. Supernatants of highly purified spermatozoa did not contain detectable levels of IL-1beta, IL-6 and VEGF. Screening of seminal plasma revealed concentrations >10-fold above the serum level for TGF-beta1, IL-8 and VEGF. Incubation of epithelial cells with 0.1, 1 and 10% seminal plasma resulted in concentration-dependant stimulation of IL-1beta, IL-6 and LIF mRNA expression. Maximum stimulation was found in epithelial cells from tissue samples taken in the mid secretory phase. Epithelial mRNA expression of IL-1beta, IL-6 and LIF increased by stimulation with TGF-beta1 and IL-8, but not with VEGF. In conclusion, seminal plasma stimulates expression of pro-inflammatory cytokines in endometrial epithelial cells in vitro. This effect might at least in part be exerted by TGF-beta1 and IL-8, abundantly present in seminal plasma. The in-vivo physiological relevance of these in-vitro studies remains to be determined.

Effect of intrauterine contraceptive devices on cytokine messenger ribonucleic acid expression in the human endometrium

To compare cytokine messenger RNA (mRNA) expression in the endometrium exposed to an intrauterine contraceptive device (IUD) and in normal cycling endometrium. Quantitative reverse-transcribed polymerase chain reaction was used to assess interleukin (IL) 1 beta, IL-6, tumor necrosis factor alpha (TNF-alpha), and colony-stimulating factor 1 (CSF-1) mRNA expression in endometrial tissue samples obtained by curettage or at hysterectomy from 11 women using a copper-releasing IUD, 10 women having a levonorgestrel-releasing IUD, and 13 fertile women during different phases of the menstrual cycle. The mRNAs encoding the studied cytokines were detected in all endometrial samples. The mean IL-1 beta, IL-6, and TNF-alpha mRNA levels were higher in the late secretory menstrual phase compared with the proliferative early secretory phase of the menstrual cycle both in endometria exposed to the copper IUD and in the control samples. The IL-1 beta and TNF-alpha mRNA levels were significantly higher in endometria exposed to the copper IUD compared with the control endometria in the late secretory menstrual phase, whereas no difference was found in the expression of these cytokine mRNAs during the proliferative early secretory phase. In contrast, the mean IL-6 mRNA level was higher in the copper IUD-exposed endometria than in the control endometria in the proliferative early secretory but not in the late secretory menstrual phase. In the levonorgestrel IUD-exposed endometria, the mean levels of IL-1 beta, TNF-alpha, and CSF-1 mRNA were similar to those in normal endometrium in the late secretory menstrual phase. The mean IL-6 mRNA level in levonorgestrel IUD-exposed endometria was lower than that in late secretory menstrual phase of controls but, because of a great individual variation in the control samples, the difference did not reach significance. No significant cyclic change nor any difference in relative CSF-1 mRNA levels between the three study groups was detected. These data provide the first evidence that the use of IUDs is associated with alterations in endometrial cytokine expression and that the alterations differ depending on the cytokine and the type of IUD. We speculate that cytokines are involved in intrauterine contraceptive effects of IUDs.