Causal relationship between OHSS and immune cells: A Mendelian randomization study.

OBJECTIVE: To confirm the causal relationship between immune cells and Ovarian Hyperstimulation Syndrome. DESIGN: Obtaining data, collecting single nucleotide polymorphisms, detecting instrumental variables heterogeneity, assessing causality, and assessing bidirectional causality. SUBJECTS: A two sample Mendelian study to confirm the causal relationship between immune cells and Ovarian Hyperstimulation Syndrome. EXPOSURE: Immune cell phenotype (including 22 million SNPs from GWAS on 3757 European individuals). MAIN OUTCOME MEASURES: Inverse variance weighting, one-sample analysis, MR-Egger, weighted median and weighted mode are used to assess the causal relationship between 731 immunophenotypes and Ovarian Hyperstimulation Syndrome. The weighted median and Mendelian Randomization multi-effect residuals and Mendelian Randomization multi-effect residuals and outlier tests are used to assess bidirectional causality between this two. RESULTS: After False Discovery Rate correction, 9 immunophenotypes were found to be significantly associated with the risk of Ovarian Hyperstimulation Syndrome. B cell panel: IgD+ AC (OR, 0.90) 、CD19 on CD24+ CD27+ (OR, 0.86) 、BAFF-R on CD20- CD38 (OR, -1.22); Mature T cell group panel: EM DN (CD4 -CD8-) AC (OR, 1.46); Myeloid cell panel: Mo MDSC AC (OR, 1.13) 、CD45 on CD33br HLA-DR+ (OR, 0.87); Monocyte panel: HLA-DR on monocyte (OR, 0.86) 、CCR2 on CD14+ CD16+ monocyte (OR, 1.15) 、cDC panel: HLA-DR on myeloid DC (OR, 0.89). CONCLUSION: This study shows the potential link between OHSS and immune cells by genetic means, providing new ideas for future clinical and basic research.

Association of IL-17 and IL-23 follicular fluid concentrations and gene expression profile in cumulus cells from infertile women at risk for ovarian hyperstimulation syndrome.

This study determined the association between the levels of interleukin (IL)-17 and IL-23 in follicular fluid (FF), as well as their mRNA levels in cumulus cells from infertile women at risk for ovarian hyperstimulation syndrome (OHSS). In this case-controlled study, the control group (n = 40) was infertile women whose partners had male factor infertility, whereas the case group (n = 40) was infertile women at risk of OHSS. IL-17 and IL-23 concentrations in FF were measured using an enzyme-linked immunosorbent assay method, whereas the mRNA expression levels of IL-17 and IL-23 of cumulus cells were determined using RT-PCR. Significantly higher levels of IL-17 were seen in the case group (p = 0.04), whereas there was no significant difference in IL-23 concentrations between the two groups (p = 0.3). The mRNA levels of IL-17 and IL-23 showed no significant differences. In the case group, there was a positive significant correlation between the IL-23 concentration in FF and the oocyte maturation rates (p = 0.01). In the case group, the number of follicles, MII oocytes, immature oocytes, fertilized oocytes and number of embryos were significantly higher than the control group (p < 0.05). Our findings showed that the mRNA expressions of IL-17 and IL-23 were similar in the two groups, and IL-17 was increased in the case group.

Immunological and hemostasiological disorders in women with ovarian hyperstimulation syndrome.

To assess the markers of destabilization of homeostasis in women with ovarian hyperstimulation syndrome (OHSS), the investigation of the levels of cortisol, markers of renin-angiotensin-aldosterone system, endothelin, proinflammatory cytokines, acute phase proteins, and parameters of hemostasis was performed. Our survey involved 105 women who became pregnant after IVF: 21 women with symptoms of the early moderate and severe OHSS, 28 women with the late moderate and severe OHSS, and 56 pregnant women undergoing IVF without symptoms of OHSS. It was found significant increase of levels of cortisol, interleukins, the number of leucocytes, concentration of fibrinogen and D-dimers in patients with early and late OHSS. The development of late OHSS is associated with the lower level of IL-8 and ceruloplasmin. The OHSS is characterized by leukocytosis, higher level of IL-6, TNF-α, fibrinogen, D-dimers, thus reflecting the homeostasis imbalance. The determination of the level of fibrinogen, D-dimers, leukocytes can be an important screening test of the intensity of the inflammatory process in patients with OHSS.

Severe ovarian hyperstimulation syndrome in patients with autoimmune disorders: a report of two cases.

BACKGROUND: Ovarian hyperstimulation syndrome (OHSS) following ovulation induction is common and varies in severity from mild to severe. Severe OHSS can be life threatening and requires hospitalization and procedures to reduce the extracellular fluid accumulation. Ovulation induction regimen and patient characteristics such as young age and low body weight have been identified as risk factors for OHSS, and follicle count >20 may be predictive of increased risk for moderate to severe OHSS. Whether any other patient factors are important in this risk equation is unknown. CASES: We report on 2 patients with type 1 diabetes mellitus (T1DM) and other autoimmune diseases who experienced a total of 4 episodes of OHSS, 2 of which were severe. CONCLUSION: T1DM, with or without other concomitant autoimmune diseases, may increase the risk for severe OHSS. Awareness of the interaction of preexisting conditions and risk for vascular leak disorders is needed for comprehensive counseling prior to embarking on ovarian stimulation. Preventive measures may be important considerations in this population.

Impaired outcome of controlled ovarian hyperstimulation in women with thyroid autoimmune disease.

BACKGROUND: Controlled ovarian hyperstimulation (COH) is a crucial step of assisted reproductive technology (ART). Thyroid dysfunction and autoimmune thyroid disease (ATD) may negatively affect the outcome of ART, but the underlying mechanisms are still poorly understood. Our aim was to evaluate the respective role of ATD and thyroid function, as assessed by serum thyrotropin (TSH), on the early outcome of COH. METHODS: In total, 262 (202 ATD-negative and 60 ATD-positive) euthyroid subfertile women underwent ART. Before COH, serum follicle-stimulating hormone (FSH), luteinizing hormone, and estradiol (E2) were measured at cycle day 3, and progesterone at cycle day 21. At oocyte pickup and at embryo transfer, we evaluated the performance of recombinant FSH (r-FSH), as assessed by serum E2 concentration/total administered r-FSH units (E2/r-FSH) ratio and by oocyte quality. RESULTS: At both oocyte pickup and embryo transfer, the performance of r-FSH was significantly poorer in ATD-positive than in ATD-negative women. In the ATD-positive group, women with a TSH <2.5 mIU/L displayed a higher serum E2 concentration at oocyte pickup, a higher E2/r-FSH ratio, and a greater number of mature metaphase II oocytes than women with a TSH >2.5 mIU/L. When ATD-positive women were divided into quartiles according to their serum TSH level, both the ovarian response to r-FSH and the number of mature metaphase II oocytes significantly increased from the lowest to the highest quartiles of serum TSH concentration. CONCLUSIONS: ATD has a negative effect on the early outcome of COH, but this negative influence may be avoided with adequate levothyroxine therapy aimed at keeping TSH <2.5 mU/L. Thyroid antibodies and serum TSH should be checked in any woman undergoing ART.

The role of IL-6 trans-signaling in vascular leakage: implications for ovarian hyperstimulation syndrome in a murine model.

CONTEXT: The inflammatory cytokine IL-6 is related to ovarian hyperstimulation syndrome (OHSS), although the functional role of IL-6 in OHSS remains largely unknown. OBJECTIVE: A key feature of the IL-6 response is that its regulation is dependent on IL-6 trans-signaling via soluble IL-6 receptor-α (sIL-6Rα). The objective of the study was to elucidate the mechanistic role of IL-6 trans-signaling in the vascular leakage that underlies the pathophysiology of OHSS. DESIGN: Ovarian endothelial cells (ECs) and granulosa-lutein cells were obtained from women undergoing in vitro fertilization. OHSS was induced in mice by administering gonadotropins for 2 days followed by human chorionic gonadotropin. The functional role of IL-6 trans-signaling in OHSS was verified using the designer cytokines Hyper IL-6 and sgp130-Fc. RESULTS: The follicular fluid levels of sIL-6Rα were elevated in women at high risk for OHSS. In the murine OHSS model, stimulation with gonadotropins significantly induces ovarian IL-6 and sIL-6Rα expression. In vitro, FSH induces de novo sIL-6Rα synthesis in granulosa-lutein cells through a protein kinase C-dependent pathway. In addition, sIL-6Rα was released by leukocytes in the presence of conditioned medium from human chorionic gonadotropin-treated granulosa-lutein cells. Ovarian ECs responded to the IL-6Rα-IL-6 complex (Hyper IL-6) but not to IL-6 alone. With activation of signal transducer and activator of transcription 3 (STAT3) and ERK, Hyper IL-6 increased vascular endothelial growth factor expression and the vascular permeability of ECs. Selective blockade of IL-6 trans-signaling by sgp130-Fc significantly inhibited vascular endothelial growth factor expression and prevented OHSS in mice. CONCLUSIONS: IL-6 trans-signaling is activated during the ovarian stimulation process. Our findings provide insight into the biologic effects of IL-6 trans-signaling in OHSS and highlight that IL-6 trans-signaling can induce vascular leakage in this disease.

Vascular endothelial growth factor production by circulating immune cells is elevated in ovarian hyperstimulation syndrome.

BACKGROUND: Ovarian hyperstimulation syndrome (OHSS) is an iatrogenic disease manifesting itself by ovarian enlargement and massive ascites with increased peritoneal capillary permeability. Although vascular endothelial growth factor (VEGF) is considered to play the main role in developing OHSS, its precise mechanism remains unclear. In this study, we examined possible roles of circulating immune cells in the pathogenesis of OHSS. METHODS: Peripheral blood mononuclear cells (PBMC) and plasma were collected from healthy non-pregnant volunteers and from patients receiving ovulation induction for IVF. PBMC were cultured for 48 h. Plasma and/or medium concentrations of VEGF, estradiol and progesterone were measured using enzyme-linked immunosorbent assay and radioimmunoassay kits. RESULTS: VEGF production by cultured PBMC and plasma concentrations of VEGF taken from patients with early onset OHSS (n = 12) were significantly higher than those in non-pregnant volunteers and patients without OHSS whose oocyte retrieval rates were similar to that of OHSS patients. OHSS patients were further classified into a high plasma VEGF concentration group and a high culture medium VEGF group. There was no significant correlation among VEGF production by PBMC and plasma concentration of VEGF, estradiol or progesterone. CONCLUSION: Although mechanistic evidence has not been provided, our study does provide new evidence to suggest that circulating immune cells are involved in the pathogenesis of OHSS via VEGF production.

Catastrophic antiphospholipid syndrome related to severe ovarian hyperstimulation.

Antiphospholipid syndrome (APS) is a cause of infertility and fetal loss. Ovarian stimulation can induce previously unknown APS. Ovarian hyperstimulation syndrome (OHS) is uncommon but potentially life-threatening, as well as catastrophic APS. A woman that simultaneously developed a severe OHS and a catastrophic APS is described in this paper. Both entities produced thrombotic cardiac and brain thrombosis. A peculiar mechanism of cardiac ischemia is also described. In spite of the life-threatening risk of this situation, the indication for preventive anti-aggregation and/or anticoagulation is not clear.

Ectopic mesothelial cell proliferation in cervical lymph nodes after severe ovarian hyperstimulation syndrome.

OBJECTIVE: To report a case of ectopic mesothelial proliferation in cervical lymph nodes after severe ovarian hyperstimulation syndrome. DESIGN: Case report. SETTING: University-affiliated teaching hospital. PATIENT(S): A 42-year-old woman underwent a successful IVF attempt and developed severe ovarian hyperstimulation syndrome and pathologically enlarged cervical lymph nodes. INTERVENTION(S): Paracentesis, cervical lymph node biopsy followed by cytology, histology, and immunohistochemistry. MAIN OUTCOME MEASURE(S): Resolution of symptoms, pregnancy outcome, correct pathological diagnosis. RESULT(S): Paracentesis resulted in resolution of symptoms of ovarian hyperstimulation. The diagnosis of ectopic mesothelial proliferation in cervical lymph nodes was made after immunohistochemical examination of cervical lymph nodes. The pregnancy progressed normally, and at 40 weeks the patient spontaneously delivered a healthy son weighing 3,060 g. CONCLUSION(S): This case describes ectopic mesothelial cell proliferation localized in and around cervical lymph nodes occurring at 9 weeks' gestation in a patient who earlier developed severe ovarian hyperstimulation syndrome 10 days after ET. Ectopic mesothelial cell proliferation in lymph nodes is an extremely rare event, and this is the first case described after ovarian hyperstimulation. Familiarity with this event is important for the clinician as well as for the pathologist in preventing the misdiagnosis of malignancy.

Interleukin-18 levels correlate with severe ovarian hyperstimulation syndrome.

OBJECTIVE: To assess the involvement of interleukin-18 (IL-18) in the pathophysiology of severe ovarian hyperstimulation syndrome (OHSS), and study its use as a marker of disease and its correlation to capillary hyperpermeability. DESIGN: Prospective controlled study. SETTING: An IVF unit in a tertiary medical center. PATIENT(S): Twenty-four patients with OHSS in an IVF program, two control groups: group 1, 40 healthy age-matched women without ovulation-induction treatment; group 2, 19 women who received the same ovulation-induction regimen without experiencing OHSS. INTERVENTION(S): Blood samples were obtained at three times: during acute OHSS, on significant clinical improvement, after complete resolution. Ascitic and pleural fluids were obtained by therapeutic paracentesis. Serum, peritoneal, and pleural fluids were analyzed for IL-18 and IL-6, and blood for hematocrit, white blood cell count, and E(2) levels. MAIN OUTCOME MEASURE(S): Hematocrit white blood cell count, serum, peritoneal, pleural fluid levels of IL-18, IL-6, E(2) in severe OHSS. RESULT(S): Significantly higher IL-18 levels were detected in serum, peritoneal, and pleural fluids of patients with severe OHSS as compared with both control groups. Serum IL-18 dropped significantly on transition to the diuretic phase and resolution. A statistically significant correlation between serum IL-18 and hyperpermeability characteristics (white blood cell count, hematocrit), serum E(2), and IL-6 levels was recorded. CONCLUSION(S): This is the first study suggesting a role of IL-18 as a marker of OHSS, with correlation to capillary hyperpermeability parameters.

Dynamic changes of the immunoglobulins in patients with severe ovarian hyperstimulation syndrome: efficacy of a novel treatment using peritoneo-venous shunt.

PROBLEM: To evaluate the efficacy of continuous auto-transfusion system of ascites (CATSA) for the treatment of patients with severe ovarian hyperstimulation syndrome (OHSS) at the risk of febrile morbidity, the dynamic changes of immunoglobulins in the sera and the peritoneal fluid from patients with severe OHSS treated by CATSA were estimated. METHOD OF STUDY: Ten patients with severe OHSS after superovulation for in vitro fertilization-embryo transfer (IVF-ET) were treated by CATSA. Immunoglobulin concentrations were examined in the serum and in the peritoneal fluid before and after CATSA. As controls, serum samples from 15 infertile women, who did not develop OHSS after the same superovulation protocol, were obtained on the day of mid-luteal period (Control-1). Serum samples from 15 patients with OHSS, who were treated by albumin infusion without paracentesis, were also obtained before and after the treatment (Control-2). RESULTS: Before the treatments, serum immunoglobulin G (IgG) concentrations in patients with severe OHSS treated with CATSA and those in patients of Control-2 were significantly lower than those in patients of Control-1 (P < 0.01). Following CATSA, the concentration of IgG increased in the sera, while it decreased in the peritoneal fluid. CONCLUSIONS: Serum IgG in patients with severe OHSS exuded into their peritoneal cavity, indicating that they might be at the status of immunodeficiency and at the risk of febrile morbidity. However, non-infectious febrile morbidity attributed to endogenous pyrogenic mechanism might be considerable. It is also suggested that CATSA might be effective in improving hypoimmunoglobulinemia of the patients with severe OHSS by the peritoneo-venous shunt.

Low peripheral blood levels of the immunosuppressive cytokine interleukin 10 (IL-10) at the start of gonadotrophin stimulation indicates increased risk for development of ovarian hyperstimulation syndrome (OHSS).

Our hypothesis was that patients developing ovarian hyperstimulation syndrome (OHSS) might have a disturbed responsiveness or delayed activation of the immunosuppresive cytokine system. In a prospective cohort study, women (n=428) undergoing in vitro fertilisation (IVF) treatment were subjected to repeated blood sampling and collection of clinical data. Fifteen patients, who developed severe OHSS, were compared with matched (age, follicle numbers, pregnancy) control patients. Samples of serum and plasma were collected throughout the stimulation and up to 7 days after embryo transfer as well as during hospitalisation for OHSS. Levels of IL-4, IL-10, IL-13, oestradiol and progesterone were measured. Significantly lower levels of IL-10 were seen at the start of gonadotrophin stimulation in OHSS patients, with an increase seen after the development of OHSS. In these OHSS patients, a negative correlation between IL-10 levels and number of follicles at time of aspiration existed, but there were no correlations between steroid and IL-10 levels. Levels of IL-13 and IL-4 were low in both groups and did not change during stimulation. The lower levels of IL-10 at start of stimulation in OHSS patients, as compared with controls, may be of pathophysiological importance by allowing for an enhanced Th-1 type immune response during gonadotrophin stimulation and thereby increased and generalised inflammation. The increase in IL-10 after development of OHSS indicates that IL-10 at that time is induced in a systemic attempt to suppress the inflammation of OHSS.

Prognostic importance of serial cytokine changes in ascites and pleural effusion in women with severe ovarian hyperstimulation syndrome.

OBJECTIVE: To determine the prognostic value of various cytokine levels in ascites and pleural effusion during the evolution of severe ovarian hyperstimulation syndrome (OHSS). DESIGN: A longitudinal study. SETTING: University teaching hospital. PATIENT(S): Twenty patients with severe OHSS who required either paracentesis or thoracentesis or both from whom ascites (n = 56) or pleural effusion (n = 12) samples were obtained. Control peritoneal fluid was obtained from 20 patients undergoing ovarian stimulation for IVF. INTERVENTION(S): Abdominal paracentesis for tense ascites and thoracentesis for massive pleural effusion. Control peritoneal fluid was obtained before oocyte retrieval. MAIN OUTCOME MEASURE(S): Levels of interleukin-1 beta (IL-1 beta), interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor-alpha (TNF-alpha), vascular endothelial growth factor (VEGF), E2, and progesterone concentrations in ascites and pleural effusion. RESULT(S): Levels of VEGF and IL-6 in ascites dropped significantly during the course of OHSS and were not correlated with E2 concentrations. Levels of VEGF were significantly correlated with levels of IL-1 beta, IL-8, and TNF-alpha, as well as progesterone concentrations, hematocrit, and white blood cell counts. None of the cytokine levels measured in pleural effusion were correlated with the course of OHSS. CONCLUSION(S): These results suggest that local cytokines might be involved in the evolution of severe OHSS and possibly serve as prognostic markers for this syndrome.

The pathogenesis of ovarian hyperstimulation syndrome: in vivo studies investigating the role of interleukin-1beta, interleukin-6, and vascular endothelial growth factor.

OBJECTIVE: To evaluate systemic and ovarian changes in levels of interleukin (IL)-1beta, IL-6, and vascular endothelial growth factor (VEGF) in response to hCG administration to determine which may be the potential initiator of vascular effects and to identify the main source of the substance; to evaluate serum and follicular fluid levels of these cytokines as markers of ovarian hyperstimulation syndrome (OHSS), and to compare levels of these cytokines under basal conditions in women with normal ovulation and those with polycystic ovary syndrome (PCOS). DESIGN: Prospective controlled study. SETTING: In vitro fertilization program at the Instituto Valenciano de Infertilidad, Valencia, Spain. PATIENT(S): Women undergoing IVF, in whom the first two study objectives were analyzed, and women with normal ovulation and patients with PCOS undergoing retrieval of immature oocytes in natural cycles or cycles stimulated for IUI but cancelled during induction of ovulation, in whom the third study objective was analyzed. INTERVENTION(S): Serum was collected before and after hCG administration, and follicular fluid was collected at ovum pick-up. MAIN OUTCOME MEASURE(S): Serum and follicular fluid levels of IL-1beta, IL-6, and VEGF. RESULT(S): There was a significant increase in serum VEGF levels after hCG administration in patients who were at risk for OHSS compared with those who were not at risk for OHSS. Significantly lower VEGF levels were found in the follicular fluid of patients who were at risk; this decrease was the only useful marker to discriminate between the two groups. Moreover, both groups had similar cytokine production under basal conditions. An increase in serum E2 occurred coincident with a decrease in IL-1beta, IL-6, and VEGF in patients with PCOS. CONCLUSION(S): Vascular endothelial growth factor seems to be the mediator of hCG on the vascular tree. There was an early systemic increase in VEGF that may have significance in the development of OHSS. A decrease in the follicular fluid VEGF concentration is a valid marker to identify women in whom OHSS will develop. The pattern of cytokine release in patients with PCOS under basal conditions was not different from that in women with normal ovulation.

Controlled ovarian hyperstimulation in assisted reproduction: effect on the immune system.

OBJECTIVE: To determine how controlled ovarian hyperstimulation (COH) in assisted reproduction affects the immune system. DESIGN: A prospective, nonrandomized, case-control study. SETTING: Academic research setting. PATIENT(S): Women with regular menstrual cycles undergoing COH in an assisted reproduction program. INTERVENTION(S): Blood samples were collected in the early and late follicular phase, at the time of ovulation, and in the luteal phase during a natural cycle, and at four times during the next cycle, which included COH and IVF. MAIN OUTCOME MEASURE(S): Lymphocyte subpopulations and the differential blood count. RESULT(S): In the natural cycles, a significant increase in the total numbers of lymphocytes, B cells, natural killer cells, and CD3+HLADR+ cells was observed in the late follicular phase, whereas the T helper/T suppressor cell ratio declined. In the hyperstimulated cycles, increases were seen in the total numbers of leukocytes and neutrophils on the day of hCG administration; the number of lymphocytes, monocytes, and neutrophils was increased on the day of oocyte retrieval, and the total number of leukocytes and neutrophils increased during the luteal phase. CONCLUSION(S): Controlled ovarian hyperstimulation with hMG and simultaneous administration of a GnRH antagonist did not affect the immune system.

Ovarian hyperstimulation syndrome: a new insight into an old enigma.

OBJECTIVE: To relay the current knowledge on the interaction between cytokines, growth factors, and ovarian hyperstimulation syndrome (OHSS) and to propose a new insight into the pathogenesis of OHSS. METHODS: Major studies that have reported on the association between cytokines, growth factors, and OHSS were identified through MEDLINE searches and through the published literature. RESULTS: Several cytokines, as well as vascular endothelial growth factor, were found to have a role, either in the prediction or pathogenesis of OHSS. Moreover, intravenous immunoglobulin, which has an anticytokine nature and inhibits the secretion of various cytokines, was shown to prevent the development of OHSS. CONCLUSION: Current knowledge indicates a close interaction between cytokines, growth factors, and OHSS. Further understanding of these interactions may lead to different understanding of ovarian hyperstimulation syndrome.

The possible role of the immune system in the aetiopathogenesis of ovarian hyperstimulation syndrome.

This review examines recent evidence suggesting a role for the immune system, in particular cytokines, in the pathogenesis of ovarian hyperstimulation syndrome (OHSS). Ovarian tissue is known to contain cells capable of producing a range of immunological mediators and the concentrations of these have been shown to be elevated in serum and ascitic fluid from women with established OHSS. Available evidence points to a role for vascular endothelial growth factor and interleukin-2, possibly acting through other intermediary cytokines, in the pathogenesis of OHSS. However, each individual has a unique cytokine profile and several cytokines may share biological actions, making it difficult to interpret data on isolated cytokine concentrations from relatively small numbers of patients. Improved understanding of the role of the immune system in the development of OHSS may have implications for the prediction, prevention and management of this iatrogenic condition.

The immune system in severe ovarian hyperstimulation syndrome.

Recently, evidence has accumulated concerning the interaction between the immune and reproductive systems that results from sharing certain lymphohematopoietic cytokines and their receptors. Furthermore, several cytokines have been implicated as having a role in the prediction and pathophysiology of ovarian hyperstimulation syndrome (OHSS). The hitherto known studies concerning the role of the immune system in OHSS, together with a hypothesis regarding the role of interleukin-2 as a possible culprit of this complication, are presented.