Ascitic Interleukin-6 Research Articles

The biology of hope: Inflammatory and neuroendocrine profiles in ovarian cancer patients

IntroductionAlthough the concept of hope is highly relevant for cancer patients, little is known about its association with cancer-relevant biomarkers. Here we examined how hope was related to diurnal cortisol and interleukin-6 (IL-6), a pro-inflammatory cytokine previously associated with tumor biology and survival in ovarian cancer. Secondly, we examined whether hope and hopelessness are distinctly associated with these biomarkers. MethodParticipants were 292 high-grade ovarian cancer patients who completed surveys and provided saliva samples 4x/daily for 3 days pre-surgery to assess diurnal cortisol. Blood (pre-surgery) and ascites were assessed for IL-6. Hope and hopelessness were assessed using standardized survey items from established scales (Center for Epidemiological Studies Depression Scale; Profile of Mood States, Functional Assessment of Cancer Therapy). Two hopeless items were z-scored and combined into a composite for analysis. Regression models related these variables to nocturnal cortisol, cortisol slope, plasma and ascites IL-6, adjusting for cancer stage, BMI, age, and depression. ResultsGreater hope was significantly related to a steeper cortisol slope, β = −0.193, p = 0.046, and lower night cortisol, β = −0.227, p = 0.018, plasma IL-6, β = −0.142, p = 0.033, and ascites IL-6, β = −0.290, p = 0.002. Secondary analyses including both hope and hopelessness showed similar patterns, with distinct relationships of hope with significantly lower nocturnal cortisol β = −0.233,p = 0.017 and ascites IL-6, β = −0.282,p = 0.003, and between hopelessness and a flatter cortisol slope, β = 0.211, p = 0.031. ConclusionsThese data suggest a biological signature of hope associated with less inflammation and more normalized diurnal cortisol in ovarian cancer. These findings have potential clinical utility but need replication with more diverse samples and validated assessments of hope.

Relative Depletion of Soluble Interleukin 6 Receptors Abolished the Development of Cytokine Release Syndrome after CART19/22 and Lenalidomide Treatment for Lymphoma

Cytokine release syndrome (CRS) is among the most representative toxicities of chimeric antigen receptor T (CART) cell therapies against non-Hodgkin's lymphoma (NHL). However, the mechanism of CRS is not fully elucidated. Interleukin-6 (IL-6) has been reported as a key factor in the development of CRS. We here present a novel mechanism of CRS emphasizing a balance between IL-6 and soluble IL-6 receptors (sIL-6Rs), essential amplifiers for IL-6-mediated inflammatory responses. A 46-year-old male patient with transformed diffuse large B-cell lymphoma (Bcl-2+/C-Myc+) refractory to 3 lines of prior therapies was enrolled in our CART trial (NCT 03196830). Baseline findings included extensive disease, multiple lesions in the abdomen, bone marrow infiltration, and a large volume of chylous ascetics with malignant cells (ascitic IL-6 baseline level: 6,691 pg/ml). And the patient received a combination of CD19-BBζ CART (8 x 106 CAR+ cells/kg), CD22-BBζ CART (1 x 106 CAR+ cells/kg), and lenalidomide (25 mg, qod, Wang X, et al., Clin Cancer Res. 2018). The patient developed mild toxicities including fever, febrile, and rash on Day 3 which were completed reversed on Day 8 after supportive care without tocilizumab or steroids. Quantitative PCR analysis of the CAR transgene demonstrated rapid expansion of the CART cells in both peripheral blood and ascites. An efficacy assessment on Day 27 revealed that most lesions disappeared or significantly shrank, the ascites nearly completely disappeared, and no malignant cell in ascites or bone marrow was detected. The patient remained in remission for 10 months. The patient was graded as CRS grade 1 according to the ASBMT scale, and cytokine assays of peripheral blood at various time points before and after CART infusions revealed expected stable levels of cytokines, including IL-6, IL-1β, IL-12, IL-15, interferon-γ, monocyte chemoattractant protein 1 (MCP1), macrophage inflammatory protein 1-alpha (MIP1α), IL-8 and granulocyte-macrophage colony-stimulating factor (GM-CSF). In the ascites, however, soaring IL-6 levels were detected with a peak value of 18,516 pg/ml on Day 7, one of the highest IL-6 levels on our CART trials, whereas no compartmental CRS or other manifestation in local abdomen was observed. The trend of ascitic IL-6 coincided with that of CAR transgene numbers. Furthermore, sIL-6R levels in ascites were lower than in serum (p < 0.0001). Importantly, the ratios of IL-6:sIL-6R were 400 to 1000 fold higher in ascites compared with in serum, highlighting that sIL-6Rs were relatively depleted in the ascites. Indeed, the absolute levels of ascitic IL-6 were close to, or exceeded at some time points, those of sIL-6R, indicating that the bottleneck of the formation of IL-6/sIL-6R signaling complex might switch from IL-6 to sIL-6R in the ascites. Flow analysis demonstrated neutrophils account for 15-20% of the total cells in ascites compared with 40-75% in serum, indicating that lack of neutrophils might be a potential cause of the low sIL-6R levels. Additionally, although sgp130 levels in ascites were also lower than in serum, the absolute values were still overwhelming to those of IL-6 and sIL-6R. Lastly, the levels of MCP1, IL-8 and angiopoietin 2 were significantly elevated, and flow analysis revealed that approximately 25% of the ascitic cells were CD14+CD11b+, indicating monocytes/macrophages might be recruited and/or activated. The levels of MIP1α, GM-CSF, IL-1β, IL-12, IL-15, interferon-γ were low in ascites during the whole period. In conclusion, our data show that relative depletion of sIL-6R in ascites may completely abolish the development of local CRS in the presence of high IL-6. This is the first report to demonstrate the features of biomarkers in ascites after CART therapy, and to describe the impacts of disproportionately low sIL-6R on IL-6 signaling without drug intervention. These results imply that the unique microenvironments in exudates such as ascites may lead to distinct pathophysiological events following CART therapy, and IL-6:sIL-6R ratio should be considered as an indicative parameter in the toxicity management of CRS. Figure Disclosures No relevant conflicts of interest to declare.

Diurnal cortisol and survival in epithelial ovarian cancer

IntroductionHypothalamic-pituitary-adrenal (HPA) deregulation is commonly observed in cancer patients, but its clinical significance is not well understood. We prospectively examined the association between HPA activity, tumor-associated inflammation, and survival in ovarian cancer patients prior to treatment.Materials and MethodsParticipants were 113 women with ovarian cancer who provided salivary cortisol for three days prior to treatment for calculation of cortisol slope, variability, and night cortisol. Cox proportional hazard regression analyses were used to examine associations between cortisol and survival in models adjusting for disease stage, tumor grade, cytoreduction and age. On a subsample of 41 patients with advanced disease ascites fluid was assayed for levels of interleukin-6 (IL-6) and correlated with cortisol variables.ResultsEach cortisol measure was associated with decreased survival time, adjusting for covariates (all p<.041). A one standard deviation increase in night cortisol was associated with a 46% greater likelihood of death. Patients in the high night cortisol group survived an estimated average of 3.3 years compared to 7.3 years for those in the low night cortisol group. Elevated ascites IL-6 was associated with each cortisol measure (all r >.36, all p<.017).DiscussionAbnormal cortisol rhythms assessed prior to treatment are associated with decreased survival in ovarian cancer and increased inflammation in the vicinity of the tumor. HPA abnormalities may reflect poor endogenous control of inflammation, dysregulation caused by tumor-associated inflammation, broad circadian disruption, or some combination of these factors. Nocturnal cortisol may have utility as a non-invasive measure of HPA function and/or disease severity.

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-1β (IL-1β), interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α), 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-1β, IL-8, and TNF-α, 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.

Elevated interleukin-6 levels in the ovarian hyper stimulation syndrome: Ovarian immunohistochemical localization of interleukin-6 signal

To examine the production and immunolocalization of interleukin-6 (IL-6) in patients with the ovarian hyperstimulation syndrome. The study group consisted of patients with ovarian hyperstimulation syndrome (n = 9) from whom serum and ascites samples were obtained. The control samples used were serum (n = 10) and peritoneal (n = 16) and follicular fluids (n = 8) from healthy individuals. Follicular fluid (n = 40) and serial serum samples were also obtained from patients undergoing menotropin stimulation for in vitro fertilization (IVF) before (n = 10) and after ovulation (n = 34). Interleukin-6 measurements were performed with a sensitive immunoassay and confirmed with a bioassay. Immunohistochemical localization of IL-6 was performed with a mouse monoclonal antibody in normal premenopausal (n = 5) and postmenopausal ovaries (n = 5), as well as with cells from stimulated follicular fluid aspirates (n = 3). We found significantly higher serum and ascites IL-6 levels in ovarian hyperstimulation syndrome (mean 18.8 +/- 1.1 and 810.8 +/- 60.7 pg/mL, respectively) compared with postovulatory serum and peritoneal fluid from normal controls (mean 4.4 +/- .69 and 44.7 +/- 7.5 pg/mL, respectively) (P < .001) or serum after menotropin stimulation (13.1 +/- 1.1 pg/mL) (P < .001). At the time of ovulation, follicular fluid IL-6 levels (normal controls, mean 9 +/- 2.1 pg/mL; menotropin stimulation, mean 10.1 +/- 4 pg/mL) were higher than in preovulatory serum (normal controls, mean 4.5 +/- .8 pg/mL; menotropin stimulation, mean 6.3 +/- 1.4 pg/mL) (P < .001). Immunohistochemical localization of IL-6 revealed intense staining in corpora lutea and theca cells from large antral follicles and luteinized granulosa cells in follicular aspirates after menotropin stimulation. Interleukin-6 levels are markedly elevated in the ovarian hyperstimulation syndrome when compared with controls. The higher follicular fluid IL-6 levels seen suggest local secretion of this cytokine. Immunohistochemical correlation demonstrated IL-6 within ovarian theca cells. These findings suggest a local role for IL-6 both in normal and stimulated ovarian function. Whether IL-6 is directly responsible for the clinical manifestations of this syndrome is unclear. However, when produced in massive amounts, the pro-inflammatory effects of IL-6 may contribute to its pathogenesis and perhaps serve as a marker for the disease.

Interleukin-6 level in serum and ascites as a prognostic factor in patients with epithelial ovarian cancer.

Interleukin-6 (IL-6) is a multifunctional cytokine that can be produced by human ovarian cancer cells. Elevated IL-6 levels have been found in the serum and ascites of patients with ovarian cancer, but its role in this disease has not been clearly established. The authors studied the relationship between IL-6 levels in serum and ascites, various tumor parameters, and survival in 70 patients with newly diagnosed, untreated epithelial ovarian cancer. Ascites and serum specimens were obtained at the time of initial surgery, and IL-6 levels were determined using the B9 bioassay. All patients underwent platinum-based chemotherapy after initial surgery. The median age of the group was 62 years (range, 28-87 years), and the median follow-up time was 13 months (range, 12-59 months). Significantly higher IL-6 levels were detected in patients' ascites (median, 49,612 pg/ml [range, < 1 to 680,330 pg/ml]) compared with serum (median, 10 pg/ml [range, < 1 to 1221 pg/ml]) (P < 0.0001). IL-6 levels in ascites correlated significantly with the volume of ascites (P < 0.0001) and nearly so with the size of tumor found at initial surgery (P = 0.05). Serum and ascites IL-6 levels did not correlate statistically with overall survival time, tumor stage, grade, histologic findings, residual tumor volume after debulking, and serum CA 125 levels. Although not statistically significant, patients who responded to chemotherapy tended to have lower ascites IL-6 levels (median, 21,102 pg/ml) compared with patients who did not respond to chemotherapy (median, 40,200 pg/ml). IL-6 is present in very high amounts in the ascites of patients with epithelial ovarian cancer. IL-6 levels in ascites correlated significantly with ascites volume and initial tumor size. IL-6 levels in ascites and serum did not correlate statistically with other tumor parameters or with survival time.

Elevated levels of interleukin-6 in ascites and serum from women with ovarian hyperstimulation syndrome

To examine the role of pro-inflammatory cytokines in ovarian hyperstimulation syndrome (OHSS). In vitro laboratory study of serum, peritoneal cells isolated and fluid obtained from ascites removed in the therapeutic management of four patients with OHSS. Tertiary care referral teaching hospital. Four patients with OHSS comprised the study population. Five healthy women at the time of elective laparoscopic tubal ligation served as controls. Control serum was also obtained from healthy adult volunteers, and control peritoneal fluid (PF) was obtained from patients on peritoneal dialysis. Therapeutic paracentesis was performed on four patients with OHSS. Peritoneal cells were isolated and fluid obtained from ascites removed in the therapeutic management of the women with OHSS. Peritoneal cells were obtained by intraoperative lavage from the control women. The cells were incubated with various concentrations of Escherichia coli lipopolysaccharide (LPS) for 20 and 48 hours. Interleukin-1 beta (IL-1 beta), tumor necrosis factor (TNF), interleukin-6 (IL-6), and prostaglandin E2 (PGE2) were assayed in the incubation supernatants. The release of the three cytokines and PGE2 in response to LPS by peritoneal cells from women with OHSS was not different from the controls. However, both serum and ascitic fluid from women with OHSS contained significantly greater levels of IL-6 than control serum and PF. No significant differences in TNF levels in serum, ascitic fluid, or PF could be found by ELISA or bioassay. Increased production of IL-6, released into the peritoneal cavity and the circulation, may mediate OHSS.

Serum and ascitic fluid levels of interleukin-1, interleukin-6, and tumor necrosis factor-alpha in patients with ovarian epithelial cancer

Recent studies have shown that multiple cytokines are secreted by ovarian epithelial cancer cells. Previous studies have shown that the cancer cell lines secrete macrophage colony-stimulating factor (M-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-1 (IL-1), interleukin-6 (IL-6), and transforming growth factor-alpha (TGF-alpha). Concomitantly, the serum levels of one of the growth factors (M-CSF) was found to be significantly elevated in patients with primary ovarian cancer and in second-look patients. The authors evaluated the serum levels of IL-1 alpha, IL-1 beta, IL-6, and tumor necrosis factor-alpha (TNF-alpha) in patients with primary ovarian epithelial cancer. These levels were then compared with cytokine concentration found in normal peritoneal fluid. Enzyme-linked immunosorbent assay (ELISA) was used to determine the levels of cytokines in normal peritoneal fluid, ascites, and serum. In serum, TNF-alpha and IL-6 were significantly increased in primary ovarian cancer patients when compared with control subjects (P < 0.0001 for both cytokines). TNF-alpha and IL-6 were also significantly higher than the levels found in second-look patients (P < 0.007 for TNF-alpha, and P = 0.0002 for IL-6). The levels of IL-1 alpha and beta were not elevated in ovarian cancer. TNF-alpha in the ascites was higher when compared with normal peritoneal fluid and was statistically significantly different when a cut-off point between 71-110 pg was selected (P < 0.005). The levels of IL-6 in ascites from patients with primary ovarian cancer also showed a marked increase (P < 0.0001) when compared with peritoneal fluid from control subjects. Levels of IL-1, IL-6, and TNF-alpha were determined in normal peritoneal fluid, ovarian malignant ascites, normal serum, and serum from patients with ovarian cancer. This study showed that the patients with ovarian cancer have elevated levels of IL-6 and TNF-alpha in serum and ascitic fluid. A larger study would help in evaluating the potential use of cytokines as tumor markers in ovarian cancer.

Evidence for the production of high amounts of interleukin-6 in the peritoneal cavity of patients with ascites

The ascites and serum concentrations of interleukin-6 (IL-6) and interleukin-1 (IL-I) were determined in 21 patients with hepatic ascites and in 9 patients with malignancy-associated ascites. There was no evidence for bacterial peritonitis in any patients. All ascites samples contained high amounts of immunoreactive IL-6 [hepatic ascites 1730 ± 2130 pg/ ml (mean ± SD), 1160 pg/ml (median); malignant ascites 4020 ± 1510 pg/ml (mean), 3820 pg/ml (median)] but no IL-1. The mean ascites to serum ratios of IL-6 were 96 (median 49) in patients with hepatic ascites and 587 (median 480) in patients with malignant ascites. Ascites IL-6 was biologically active as determined by the B9 cell bioassay. The results indicate that even in the absence of infection IL-6 is produced in high amounts in the peritoneal cavity of patients with hepatic or malignant ascites.