Levels Of sTNFRs Research Articles

Abstract 4426: Removal of soluble TNF receptors as a novel form of immunotherapy for patients with advanced solid tumors

Abstract Immunicom, Inc. has developed a novel subtractive immunotherapeutic apheresis method (Immunopheresis®) to selectively remove soluble tumor necrosis factor receptors, sTNF-R1 and sTNF-R2 (sTNF-Rs) generated by tumors. sTNF-Rs block the tumoricidal activities, hence their removal restores the anticancer immunotherapeutic activity of native TNF-α. Past use of systemic administration of TNF-α to treat solid tumors resulted in unacceptable toxicities. Immunicom’s LW-02 Column contains a bead matrix covalently bonded to a single-chain, TNF-α capture ligand (scTNF) with high affinity and selectivity to remove sTNF-Rs from plasma. Three clinical trials (NCT04004910, NCT04142931, NCT04690686) have been conducted in patients with various advanced-stage breast cancers (BC), melanoma (M), renal cell carcinoma (RCC) and non-small cell lung cancers (NSCLC). Patients were assigned to receive LW-02 Column Immunopheresis® monotherapy or combined with various chemo- or immunotherapies. Immunopheresis® was administered 3x/week for each trial’s 12 or 16-week Primary Treatment period with each procedure processing 2 plasma volumes (2PV). Patients with (at least) clinical stability and/or tumor responses could receive additional treatment. Compared to healthy persons, patients in all 3 trials were noted as having significantly elevated baseline blood concentrations of sTNF-R1 and sTNF-R2. For instance, sTNF-R1 plasma levels were 3.5 ng/mL, 3.5 ng/mL, 2.2 ng/mL and 2.1 ng/mL for BC, M, RCC and NSCLC, respectively, which were 5-8-fold higher than normal values. Likewise, sTNF-R2 plasma concentrations (which can be 4-fold, or more, greater than sTNF-R1) were 4-9-fold higher for BC (16.5 ng/mL), M (19.1), RCC (22.7) or NSCLC (11.1 ng/mL) than normal values. Through the data cutoff, >1,600 LW-02 Column Immunopheresis® procedures were performed. On average, 95% of sTNF-R1 and 80% of sTNF-R2 are removed from plasma at the 30-minute procedure timepoint. At the 2PV timepoint, overall mean reductions of sTNF-R1 and sTNF-R2 in whole blood were 51 and 52%. Further, minimal leaching (mean of 113 ng per procedure), of the scTNF capture ligand was detected, a quantity that is insufficient to cause any meaningful clinical effects. In vitro testing determined that the LW-02 Column has no discernable evidence of unintended or “off-target” removal of other cytokines or blood constituents or any non-specific binding. Moreover, the trial results demonstrate Immunopheresis® is safe with minimal side effects determined to be related to treatment with the column. Results from 3 trials indicate that cancer patients with advanced solid tumors present with significantly upregulated levels of immune inhibitory sTNF-Rs. LW-02 Column Immunopheresis® can safely and selectively remove sTNF-Rs thus allowing endogenous TNF-α to instigate its multiple anticancer pathways. Citation Format: Piotr J. Wysocki, Piotr Tomczak, Tomasz Jankowski, Tomasz Nowikiewicz, Gal Markel, Ronnie Shapira, Gokhan Demir, Mustafa Bozkurt, Sameera Bilgrami, Annette Marleau, Adam Ostrowski, Lawrence Florin, Victoria Manax, Robert Segal. Removal of soluble TNF receptors as a novel form of immunotherapy for patients with advanced solid tumors. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4426.

Clinical and molecular correlates of the ASPECTS in the acute phase of stroke.

The Alberta Stroke Program Early CT Score (ASPECTS) scale was developed for monitoring early ischemic changes on CT, being associated with clinical outcomes. The ASPECTS can also associate with peripheral biomarkers that reflect the pathophysiological response of the brain to the ischemic stroke. To investigate the association between peripheral biomarkers with the Alberta Stroke Program Early CT Score (ASPECTS) in individuals after ischemic stroke. Patients over 18 years old with acute ischemic stroke were enrolled in this study. No patient was eligible for thrombolysis. The patients were submitted to non-contrast CT in the first 24 hours of admission, being the Alberta Stroke Program Early CT Score and clinical and molecular evaluations applied on the same day. The National Institutes of Health Stroke Scale (NIHSS), modified Rankin scale and the Mini-Mental State Examination for clinical evaluation were also applied to all subjects. Plasma levels of BDNF, VCAM-1, VEGF, IL-1β, sTNFRs and adiponectin were determined by ELISA. Worse neurological impairment (NIHSS), cognitive (MEEM) and functional (Rankin) performance was observed in the group with changes in the NCTT. Patients with NCTT changes also exhibited higher levels of IL-1β and adiponectin. In the linear multivariate regression, an adjusted R coefficient of 0.515 was found, indicating adiponectin and NIHSS as independent predictors of ASPECTS. Plasma levels of adiponectin are associated with the ASPECTS scores.

Analysis of biomarker serum levels in IVIG and infliximab refractory Kawasaki disease patients.

Infliximab (IFX) is effective for treatment of refractory Kawasaki disease (KD). However, the precise mechanisms and biomarkers for IFX efficacy are unknown. We tried to evaluate the effect and response to IFX therapy by measuring serum cytokine levels. Twenty-nine children with KD who had been resistant to two courses of high-dose intravenous immunoglobulin were enrolled and treated with IFX. Plasma samples were analyzed for cytokines before and after IFX administration. Serum levels of interleukin-6, granulocyte colony-stimulating factor (G-CSF), interferon-gamma-induced monokine, interferon-gamma inducible protein 10 (IP-10), monocyte chemotactic protein 1, and soluble tumor necrosis factor-alpha receptor (sTNFR) 1 and 2 were significantly elevated before IFX treatment, but promptly decreased after the administration. The pre-treatment G-CSF and sTNFR1 levels in non-responders to IFX were significantly higher than in responders, who were defined as patients who defervesce (< 37.5°C). After IFX administration, elevated cytokines declined to normal ranges in responders, but in non-responsive group, G-CSF and sTNFR1 remained elevated without failing to normal levels. IFX treatment significantly reduced the levels of serum cytokines, chemokines, and sTNFRs in refractory KD. G-CSF and sTNFR1 may be indicators predictive of poor response to IFX.

Correlation of higher levels of soluble TNF-R1 with a shorter survival, independent of age, in recurrent glioblastoma.

The circulating levels of soluble tumor necrosis factor receptor-1 (sTNF-R1) and sTNF-R2 are altered in numerous diseases, including several types of cancer. Correlations with the risk of progression in some cancers, as well as systemic manifestations of the disease and therapeutic side-effects, have been described. However, there is very little information on the levels of these soluble receptors in glioblastoma (GBM). Here, we report on an exploratory retrospective study of the levels of sTNF-Rs in the vascular circulation of patients with GBM. Banked samples were obtained from 112 GBM patients (66 untreated, newly-diagnosed patients and 46 with recurrent disease) from two institutions. The levels of sTNF-R1 in the plasma were significantly lower in patients with newly-diagnosed or recurrent GBM than apparently healthy individuals and correlated with the intensity of expression of TNF-R1 on the tumor-associated endothelial cells (ECs) in the corresponding biopsies. Elevated levels of sTNF-R1 in patients with recurrent, but not newly-diagnosed GBM, were significantly associated with a shorter survival, independent of age (p = 0.02) or steroid medication. In contrast, the levels of circulating sTNF-R2 were significantly higher in recurrent GBM than healthy individuals and there was no significant correlation with expression of TNF-R2 on the tumor-associated ECs or survival time. The results indicate that larger, prospective studies are warranted to determine the predictive value of the levels of sTNF-R1 in patients with recurrent GBM and the factors that regulate the levels of sTNF-Rs in the circulation in GBM patients.

Associations Between Serum Inflammatory Markers and Hippocampal Volume in a Community Sample.

To quantify associations between inflammatory biomarkers and hippocampal volume (HV) and to examine effect modification according to sex, race, and age. Cross-sectional analyses using generalized estimating equations to account for familial clustering; standardized β-coefficients adjusted for age, sex, race, and education. Community cohorts in Jackson, Mississippi and Rochester, Minnesota. The Genetic Epidemiology Network of Arteriopathy study. C-reactive protein (CRP), interleukin-6 (IL-6), and soluble tumor necrosis factor receptors 1 (sTNFR-1) and 2 (sTNFR-2) from peripheral blood were measured in a sample of 773 non-Hispanic whites (61% women, aged 60.2 ± 9.8) and 514 African Americans (70% women, aged 63.9 ± 8.1) who also underwent brain magnetic resonance imaging. Biomarkers were standardized and compared according to sex, race and age with HV. In the full sample, higher sTNFR-1 and sTNFR-2 were associated with smaller HV. Each standard deviation (SD) increase in sTNFR-1 was associated with 59.1 mm(3) (95% confidence interval (CI) = -101.4 to -16.7 mm(3) ) smaller HV and each SD increase in sTNFR-2 associated with 48.8 mm(3) (95% CI = -92.2 to -5.3 mm(3) ) smaller HV. Relationships were stronger for sTNFR-2 in men (HV = -116.6 mm(3) for each SD increase, 95% CI = -201.0 to -32.1) than women (HV = -26.0 per SD increase, 95% CI = -72.4-20.5) and sTNFR-1 in non-Hispanic whites (HV = -84.7 mm(3) per SD increase, 95% CI = -142.2 to -27.1) than African Americans (HV = -14.1 mm(3) per SD increase, 95% CI = -78.3-50.1). Associations between IL-6 or CRP and HV were not supported. Higher levels of sTNFRs were associated cross-sectionally with smaller hippocampi. Longitudinal data are needed to determine whether these biomarkers may help to identify risk of late-life cognitive impairment.

Soluble tumor necrosis factor receptor 1 is associated with diminished estimated glomerular filtration rate in colombian patients with type 2 diabetes

AimsThe tumor necrosis factor α (TNF-α) family of inflammatory molecules plays a crucial role in the pathogenesis of type 2 diabetes mellitus (DM2) complications. TNF-α soluble receptors 1 (sTNFR1) and 2 (sTNFR2) have been associated with chronic kidney disease in DM2 patients. This cross-sectional study intended to determine serum concentrations of sTNFR1 and sTNFR2 in Colombian patients and correlated them with various clinical variables, especially kidney function. Methods92 Colombian patients with DM2 were recruited. Anthropometric variables, glycemic control parameters, lipid profile and renal function were assessed for each patient. Levels of sTNFR1 and sTNFR2 were determined using ELISA. Patients were stratified in two groups according to reduced estimated glomerular filtration rate (eGFR) (<60ml/min/1.73m2) and normal eGFR (≥60ml/min/1.73m2). ResultsSignificantly elevated levels of sTNFR1 and sTNFR2 were observed in the diminished versus normal eGFR group. Also, significant differences were noticed between both groups in haemoglobin A1c (HbA1c) values, percentage of hypertensive subjects treated with angiotensin receptor blocker (ARB) and subjects treated with metformin. No differences were observed regarding body mass index (BMI), albuminuria and lipid profile. Multivariable linear regression analysis revealed that sTNFR1 alone showed a significant association with low eGFR (p=0.009). However, after adjusting for age, the association weakens. Moreover, sTNFR1 and sTNFR2 showed a linear negative correlation with eGFR (r=–0.448, p<0.001 and r=–0.376, p<0.001, respectively). A positive correlation was also seen between sTNFR1 and HbA1c, whereas a negative correlation between both sTNFRs and high-density lipoprotein (HDL) cholesterol was found. ConclusionElevated levels of sTNFRs, especially sTNFR1, are associated with loss of kidney function in Hispanic patients with DM2. Future studies should focus on social and genetic determinants of inflammation and their association with CKD in this ethnicity.

Association of soluble tumor necrosis factor receptors 1 and 2 with nephropathy, cardiovascular events, and total mortality in type 2 diabetes

Aims/hypothesisSoluble tumor necrosis factor receptors 1 and 2 (sTNFR1 and sTNFR2) contribute to experimental diabetic kidney disease, a condition with substantially increased cardiovascular risk when present in patients. Therefore, we aimed to explore the levels of sTNFRs, and their association with prevalent kidney disease, incident cardiovascular disease, and risk of mortality independently of baseline kidney function and microalbuminuria in a cohort of patients with type 2 diabetes. In pre-defined secondary analyses we also investigated whether the sTNFRs predict adverse outcome in the absence of diabetic kidney disease.MethodsThe CARDIPP study, a cohort study of 607 diabetes patients [mean age 61 years, 44 % women, 45 cardiovascular events (fatal/non-fatal myocardial infarction or stroke) and 44 deaths during follow-up (mean 7.6 years)] was used.ResultsHigher sTNFR1 and sTNFR2 were associated with higher odds of prevalent kidney disease [odd ratio (OR) per standard deviation (SD) increase 1.60, 95 % confidence interval (CI) 1.32–1.93, p < 0.001 and OR 1.54, 95 % CI 1.21–1.97, p = 0.001, respectively]. In Cox regression models adjusting for age, sex, glomerular filtration rate and urinary albumin/creatinine ratio, higher sTNFR1 and sTNFR2 predicted incident cardiovascular events [hazard ratio (HR) per SD increase, 1.66, 95 % CI 1.29–2.174, p < 0.001 and HR 1.47, 95 % CI 1.13–1.91, p = 0.004, respectively]. Results were similar in separate models with adjustments for inflammatory markers, HbA1c, or established cardiovascular risk factors, or when participants with diabetic kidney disease at baseline were excluded (p < 0.01 for all). Both sTNFRs were associated with mortality.Conclusions/InterpretationsHigher circulating sTNFR1 and sTNFR2 are associated with diabetic kidney disease, and predicts incident cardiovascular disease and mortality independently of microalbuminuria and kidney function, even in those without kidney disease. Our findings support the clinical utility of sTNFRs as prognostic markers in type 2 diabetes.

Plasma levels of soluble TNF receptors 1 and 2 after tDCS and sertraline treatment in major depression: Results from the SELECT-TDCS trial

BackgroundThe cytokine hypothesis of depression postulates that the pathophysiology of this illness incorporates an increased production of pro-inflammatory cytokines, which leads to an over-activation of the hypothalamic–pituitary–adrenal axis as well as monoaminergic disturbances. Nevertheless, it remains unclear whether the amelioration of depressive symptoms could decrease cytokine levels. Notwithstanding antidepressant drug therapy might exert anti-inflammatory effects, the effects of non-invasive neuromodulatory approaches like transcranial direct current stimulation (tDCS) on pro-inflammatory cytokine networks are largely unknown. MethodsWe evaluated, in the Sertraline vs. Electric Current Therapy for Treating Depression Clinical Study (SELECT-TDCS) trial, whether the plasma levels of the soluble TNF receptors 1 and 2 (sTNFRs) changed after antidepressant treatment in a sample of 73 antidepressant-free patients with unipolar depressive disorder in an episode of at least moderate intensity. ResultsAlthough both tDCS and sertraline exerted antidepressant effects, the plasma levels of sTNFRs did not change over time regardless of the intervention and clinical response. Also, baseline sTNFRs levels did not predict antidepressant response. LimitationsOur negative findings could be a type II error, as this trial did not use an equivalence design. ConclusionsTo conclude, in this novel placebo-controlled trial prospectively evaluating the changes of sTNFRs in depressed patients, we found that these molecules are not surrogate biomarkers of treatment response of tDCS, whose antidepressant effects occurred regardless of normalization of immunological activity.

High Levels of Soluble Tumor Necrosis Factor Receptors 1 and 2 and Their Association with Mortality in Patients Undergoing Hemodialysis

Objective: Circulating soluble tumor necrosis factor receptors 1 and 2 (sTNFR1 and sTNFR2) are associated with chronic kidney disease (CKD) progression in patients with CKD or diabetes, and with higher mortality. However, data in patients with end-stage renal disease are scarce. Therefore, we analyzed serum levels of sTNFR1 and sTNFR2 and investigated their association with inflammatory markers and mortality in dialysis patients. Research Design and Methods: This was a longitudinal cohort study of 207 prevalent patients (median age 66 years, 56% men) undergoing hemodialysis in Stockholm, Sweden. Demographics, clinical characteristics, including comorbidities and laboratory data, were obtained at baseline, together with prospective follow-up for mortality. Results: The median sTNFR1 and sTNFR2 levels were 17,680 ng/l [95% confidence interval (CI) 17,023-18,337] and 24,450 ng/l (95% CI 23,721-25,179), respectively. During a follow-up of 31 months (interquartile range, 21-38), 77 patients died. There was no association between the levels of sTNFRs and mortality in Cox regression models, and no consistent trend towards higher or lower mortality was seen in Laplace regression models. sTNFR1 and sTNFR2 levels were highly associated with other inflammatory markers including interleukin-6, pentraxin 3 and TNF-α. Conclusions: Prevalent hemodialysis patients have several-fold higher levels of sTNFRs compared to previous studies in CKD stage 4 patients. As no consistent association between TNFR and mortality was observed, clinical implications of measuring these receptors to predict outcome end-stage renal disease patients provide limited results.

Soluble endoglin, transforming growth factor-Beta 1 and soluble tumor necrosis factor alpha receptors in different clinical manifestations of preeclampsia.

BackgroundDespite intensive research, the etiopathogenesis of preeclampsia (PE) remains uncertain. Inflammatory and angiogenic factors are thought to play considerable roles in this disease. The objective of this study was to investigate the association between soluble endoglin (sEng), transforming growth factor beta-1 (TGF-β1) and tumor necrosis factor alpha soluble receptors (sTNF-Rs) and the clinical manifestations of PE.MethodsPlasma levels of sEng, TGF-β1 and sTNF-Rs were determined by ELISA in 23 non-pregnant, 21 normotensive pregnant and 43 PE women. PE women were stratified into subgroups according to the severity [mild (n = 12) and severe (n = 31)] and onset-time of the disease [early (n = 19) and late (n = 24)].ResultsPregnancy was associated with higher levels of sEng, sTNF-R1 and sTNF-R2 than the non-pregnant state. Moreover, PE women had higher levels of sEng and sTNF-R1 than normotensive pregnant women. No difference was found in TGF-β1 levels, comparing the three study groups. Late PE had higher levels of sTNF-R1 and sTNF-R2 than early PE. No significant differences were found in sEng and TGF-β1 comparing early and late PE. sEng levels were higher in severe PE than in mild PE and no difference was found for TGF-β1, sTNF-R1 and sTNF-R2 levels. There was a positive correlation among sEng, TNF-R1 and sTNF-2 levels. Logistic regression analysis revealed that primiparity and sEng levels are independently associated with the development of PE. Furthermore, sEng levels are independently associated with the disease severity.ConclusionsThese results suggest that pregnancy is a condition associated with higher levels of anti-angiogenic and pro-inflammatory factors than the non-pregnant state and that PE is associated with an imbalance of these factors in the maternal circulation.

Effect of six-month lifestyle intervention on adiponectin, resistin and soluble tumor necrosis factor-α receptors in obese adolescents.

The aim of this study was to evaluate the effect of a six-month lifestyle intervention on adiponectin, resistin, and two soluble forms of tumor necrosis factor-α receptor (sTNFR) in obese adolescents. A total of 54 obese adolescents aged 10 to 16 years completed the program. Twenty-four adolescents with normal weight at baseline were used as a control group. Our results demonstrated that obese adolescents had abnormal lipid profile, homeostasis model assessment (HOMA) index, adiponectin level (5.6 ± 2.7 vs. 7.6 ± 2.9 μg/mL, p = 0.005) as well as resistin level (31.0 ± 9.0 vs. 24.3 ± 8.5 ng/mL, p = 0.003), whereas levels of both sTNFRs were similar to those in normal weight subjects. After the six-month lifestyle intervention, obese adolescents had a slight but significant drop in standard deviation score-body mass index (SDS-BMI), a significant decrease in waist circumference, total cholesterol, triglycerides, HOMA index, as well as resistin, and a significant increase in adiponectin and high-density lipoprotein-cholesterol. In adolescents without decreased SDS-BMI, no change was observed in adipokines. Changes in adiponectin correlated negatively with changes in waist circumference (r = -0.275, p = 0.044). Changes in resistin correlated positively with changes in triglycerides (r = 0.302, p = 0.027). The study demonstrated the increase of resistin and the decrease of adiponectin in obese adolescents. Lifestyle intervention improved adipokine abnormalities in obese subjects.

Soluble Tumor Necrosis Factor Receptors Shed by Breast Tumor Cells Inhibit Macrophage Chemotaxis

Breast tumor cells alter their microenvironment in part through the expression of protumor molecules that influence macrophages during tumor progression and metastasis. Macrophage recruitment is stimulated by chemotactic factors, including tumor necrosis factor alpha (TNF-α), which also stimulates the cytotoxic/tumor cell killing macrophage phenotype. Through TNF-α converting enzyme (TACE/ADAM17) activities, breast tumor cells shed membrane-bound proteins, including their TNF receptors (sTNFR1/2), which serve as decoys sequestering TNF-α and preventing TNF-α-driven apoptosis of tumor cells, thereby decreasing TNF-α bioavailability. Here we investigated the levels of sTNFRs shed by breast tumor cells and determined the effects of shed sTNFRs on macrophage migration toward TNF-α. TNF-α and sTNFRs concentrations were measured in murine normal epithelial, stromal, and mammary tumor cells. The migration of murine macrophages towards TNF-α in the presence of tumor derived soluble factors (TDSFs) shed by TACE was determined. TNF-α concentrations secreted by tumor and normal epithelial cells were below the detection limit contrasting with stromal cells, especially macrophages, which expressed higher levels of TNF-α (P<0.001). Regardless of the cell tested, treatment with the TACE inhibitor TAPI-0 led to a significant decrease in sTNFR2 shed (P<0.05). The dose-dependent macrophage migration toward TNF-α prevented by incubation with TDSFs was not observed with TDSFs collected following TAPI-0 treatment (P<0.05). Furthermore, the TNF-α-driven increased pAkt expression in macrophage was inhibited by TACE shed TDSFs (P<0.05). These results highlight the role of tumor-shed sTNFRs in TNF-α -driven macrophage chemotaxis.

Combination of plasma tumor necrosis factor receptors signaling proteins, beta-amyloid and apolipoprotein E for the detection of Alzheimer's disease

Activation of inflammatory processes has been observed within the brain as well as periphery of subjects with Alzheimer's disease (AD). Among several putative neuroinflammatory mechanisms, the tumor necrosis factor-α (TNF-α) signaling system plays a central role. TNF-α converting enzyme (TACE) does not only cleave pro-TNF-α but also TNF receptors, however, whether the TACE activity and soluble TNF receptors (sTNFRs) were changed in the plasma were not clear. The aim of this study was to determine whether the levels of TACE activity and sTNFRs are sufficiently altered in the plasma of AD patients to be helpful in AD diagnosis. We examined TACE levels in the plasma of 153 patients with AD, 98 patients with amnestic mild cognitive impairment (aMCI), 53 patients with vascular dementia (VaD), and 120 age-matched healthy control subjects, and found TACE activity and sTNFRs were significantly higher in patients with AD and aMCI compared with control subjects (TACE: P<0.001, P<0.01; sTNFR1: P<0.001, P<0.001; sTNFR2: P<0.001, P<0.01, respectively). The TACE activity and sTNFRs levels in VaD patients were significantly higher than the levels observed in AD patients (TACE activity: P<0.001, sTNFR1: P<0.01, sTNFR2: P<0.01). In the plasma of AD patients, the levels of both TACE activity and sTNFRs positively correlated with the levels of Aβ40 and negatively correlated with the ratio of Aβ42/Aβ40. AD patients with at least one copy of the ApoEε4 allele showed higher TACE activity and sTNFR plasma levels compared with patients without the ApoEε4 allele. We then combined the data on plasma TACE activity, sTNFRs, and Aβ with the presence of the APOEε4 allele and found that this biomarker panel exhibited a high sensitivity and specificity for discriminating AD patients from non-demented control subjects and VaD patients.

Increased Serum Concentrations of Circulating Glycocalyx Components in HELLP Syndrome Compared to Healthy Pregnancy: An Observational Study

Severe inflammation has been shown to induce a shedding of the endothelial glycocalyx (EGX). Inflammatory cytokines, such as tumor necrosis factor α (TNF-α), impede the thickness of the EGX. While a controlled inflammatory reaction occurs already in normal pregnancy, women with hemolysis, elevated liver enzymes and low platelets (HELLP) syndrome had an exaggerated inflammatory response. This study investigates the shedding of the glycocalyx during normal pregnancy and in women with HELLP syndrome. Glycocalyx components (syndecan 1, heparan sulfate, and hyaluronic acid) were measured in serum of healthy women throughout pregnancy (4 time points, n = 26), in women with HELLP syndrome (n = 17) before delivery and in nonpregnant volunteers (n = 10). Serum concentrations of TNF-α and soluble TNF-α receptors (sTNF-Rs) were assessed once in all 3 groups. Syndecan 1 serum concentrations constantly rose throughout normal pregnancy. Immediately before delivery, a 159-fold increase was measured compared to nonpregnant controls (P < .01). Even higher amounts were observed in patients with HELLP prior to delivery (median 12252 ng/mL) compared to healthy women matched by gestational age (median 5943 ng/mL; P < .01). Relevantly, increased serum levels of heparan sulfate, hyaluronic acid, and sTNF-Rs were only detected in patients with HELLP (P < .01). These findings suggest that considerable amounts of syndecan 1 are released into maternal blood during uncomplicated pregnancy. The HELLP syndrome is associated with an even more pronounced shedding of glycocalyx components. The maternal vasculature as well as the placenta has to be discussed as a possible origin of circulating glycocalyx components.

Combination of Indoleamine 2,3-Dioxygenase Expression and L-Kynurenine Level Can Identify Truly Poor Prognosis Patients with Diffuse Large B-Cell Lymphoma

Abstract 2639 INTRODUCTION:Recent studies have suggested that one mechanism of immune escape could be catabolism of the essential amino acid tryptophan, via indoleamine 2,3-dioxygenase (IDO) along the kynurenine pathway. Some metabolites derived from tryptophan by IDO, such as L-kynurenine, block antigen-driven specific T-cell proliferation and induce T-cell death. IDO activity thus plays an important role in regulating immune responses exerted by antigen-presenting cells (APCs) and also provides transformed cells with a potent tool to help escape from assault by the immune system. We have recently been able to clarify the utility of either serum L-kynurenine or tissue IDO expression as prognostic factors for DLBCL patients treated using R-CHOP. The present study therefore examined the serum level of L-kynurenine and tissue IDO expression simultaneously in DLBCL patients in an attempt to identify patients showing truly poor prognosis. MATERIALS AND METHODS:The study protocol used a retrospective case series design that was approved by our institutional review board. We investigated DLBCL patients who had been histologically diagnosed according to the WHO classification of hematopoietic tumors between December 2003 and November 2006. All serum samples were obtained on admission before starting any treatments. Informed consent was obtained from all patients for the use of samples in this study. Tumor tissue was obtained from every patient by lymph node biopsy before initiation of therapy. Patients were assigned to receive six to eight cycles of R-CHOP or R-THP-COP therapy. Each regimen consisted of rituximab (375 mg/m2 on day 1), cyclophosphamide (750 mg/m2 on day 3), doxorubicin (DOX) or tetrahydropyranyl-adriamycin (THP) (50 mg/m2, given as a 30-min infusion on day 3), vincristine (1.4 mg/m2 on day 3, with a maximal dose of 2.0 mg), and prednisolone (100 mg/day on days 3–7). We measured serum concentrations of L-kynurenine by high-performance liquid chromatography (HPLC), and serum levels of sTNF-Rs (p55, TNF-R1; p75, TNF-R2) by ELISA. And we examined IDO expression in DLBCL tissues by immunohistochemical staining in a total of 119 patients. RESULTS:Median serum levels of L-kynurenine were significantly higher in DLBCL patients (1.598 mM; range, 0.489–5.066 mM) than in healthy volunteers (1.133 mM; range, 0.752–1.388 mM). And we found that lymphoma cells expressed IDO in 32% cases. We therefore classified cases as IDO-positive DLBCL or IDO-negative DLBCL based on whether lymphoma cells themselves expressed IDO. No significant correlation was identified between serum L-kynurenine level and IDO expression. Serum L-kynurenine level showed a significant positive correlation with concentrations of sTNF-R1 and sTNF-R2 in IDO-negative DLBCL patients. CR rates with IDO-positive and IDO-negative DLBCL were 42% and 75%, respectively (p=0.012). Three-year OS rates were 55% and 78% for patients with L-kynurenine ≥1.5 and <1.5 mM, respectively (p=0.027), and 44% and 75% for patients with IDO-positive and IDO-negative DLBCL, respectively (p=0.0017). Univariate Cox analysis identified the following prognostic factors: IDO expression, serum kynurenine, PS, and multiple site extranodal involvement. Multivariate analysis of these four parameters showed that positive IDO expression and serum kynurenine ≥1.5 mM were significant prognostic factors. We analyzed patients’ outcome according to serum concentration of L-kynurenine and IDO expression of lymphoma cells at diagnosis. Patients with low kynurenine and IDO-negative showed the best outcomes, while patients with high kynurenine and IDO-positive showed the poorest outcome. This combination distinguishes 3 separate prognostic groups of “very good”, “good”, and “poor”, with 3-year OS ranging from 84.1% to 31.1% (p<0.001, Figure). CONCLUSION:IDO activity might play an important role in the disease activity of DLBCL. The combination of tissue IDO expression and serum level of L-kynurenine is a useful tool to predict the prognosis of patients with DLBCL. [Display omitted] Disclosures:No relevant conflicts of interest to declare.

Circulating levels of TNF-α and its soluble receptors in the plasma of patients with epithelial ovarian cancer

The significance of circulating levels of TNF-alpha and its soluble receptors (sTNF-Rs) in the plasma of patients with epithelial ovarian cancer (EOC) has not been fully elucidated. The present study was to investigate the relationship of pretreatment plasma levels of TNF-alpha, sTNFR-1 and sTNFR-2 with outcome in 126 patients with EOC. Concentrations of TNF-alpha and sTNF-Rs were determined by enzyme-linked immunosorbent assay (ELISA). Median TNF-alpha and sTNF-Rs levels were significantly higher in EOC patients than in healthy controls. High plasma levels of TNF-alpha and sTNF-Rs were correlated with tumor stage and with reduced mean survival time (MST). The results of the present study suggested that preoperative plasma TNF-alpha and sTNF-Rs levels in EOC patients correlated with the highest risk of cancer progression. Thus, the clinical value of an activated TNF system in EOC needs to be further investigated.

Increased levels of immunological markers in the respiratory tract but not in serum correlate with active pulmonary mycobacterial infection in mice

Immunological tests for the diagnosis of tuberculosis (TB) have relied mostly on detection of immune markers in serum or release of cytokines by mononuclear cells in vitro. These tests, although useful, sometimes fail to discriminate between active infection and contact with mycobacteria or vaccination. TB is primarily a disease of the lung, and therefore identification of immunological markers in the respiratory tract will be more likely to reflect the infection status or disease activity. In this study, it is demonstrated that active infection of mice with Mycobacterium bovis bacille Calmette-Guérin (BCG), but not exposure to heat-killed BCG, induced production of interleukin-12 (IL-12), interferon-γ (IFN-γ) or soluble tumour necrosis factor receptors (sTNFRs) locally in the lungs, as detected in bronchoalveolar lavage (BAL) fluid. There was a strong correlation between bacterial growth in the lung and levels of sTNFRs, and to some extent IL-12 and IFN-γ, in BAL fluid. Furthermore, sTNFR levels increased significantly in BAL fluid after reactivation of controlled infection with dexamethasone, and this correlated with increased bacterial growth in the lungs. Finally, infection, but not exposure to non-replicating mycobacteria, induced specific IgG and IgA in BAL fluid. Elevated levels of all biomarkers measured were also detected in the serum, but correlation with infection was not as clear as in the case of BAL fluid. Taken together, the detection of sTNFRs and mycobacterium-specific antibodies, especially IgA, locally in the lungs could be used as immunological markers for the diagnosis of TB.

Soluble TNF receptors are associated with Aβ metabolism and conversion to dementia in subjects with mild cognitive impairment

There is evidence supporting that tumor necrosis factor receptor (TNFR)-signaling can induce production of beta-amyloid (Aβ) in the brain. Moreover, amyloid-induced toxicity has been shown to be dependent on TNFR-signaling. However, it is still unclear whether TNFRs are involved in the early stages of dementia. We analyzed soluble TNFR1 and TNFR2 levels in plasma and cerebrospinal fluid (CSF) at baseline in 137 patients with mild cognitive impairment (MCI) and 30 age-matched controls. The MCI patients were followed for 4-6 years with an incidence of Alzheimer's disease (AD) or vascular dementia (VaD) of 15% per year. The patients with MCI who subsequently developed these forms of dementias had higher levels of sTNFR1 and sTNFR2 in both CSF and plasma already at baseline when compared to age-matched controls (p<0.05). In the CSF of MCI subjects and controls the levels of both sTNFR1 and sTNFR2 correlated strongly with β-site APP-cleaving enzyme 1 (BACE1) activity (r(s)=0.53-0.68, p<0.01) and Aβ 40 levels (r(s)=0.59-0.71, p<0.001). Similarly, both sTNFRs were associated with Aβ 40 (r(s)=0.39-0.46, p<0.05) in plasma. Finally, the levels of both sTNFRs correlated with the axonal damage marker tau in the CSF of MCI subjects and controls (r(s)=0.57-0.83, p<0.001). TNFR-signaling might be involved in the early pathogenesis of AD and VaD, and could be associated with beta-amyloid metabolism.

Genetic priming of a proinflammatory profile predicts low IQ in octogenarians

The purpose of the study was to test the hypothesis that single nucleotide polymorphisms (SNPs) within interleukin (IL)-18, TNF-α, IL-6 and IL-10 gene promoter regions are risk factors for cognitive decline in healthy octogenarians, and to isolate the strongest inflammatory biomarkers of cognitive function in the peripheral blood. The Wechsler Adult Intelligence Scale was administered to 112 individuals at ages 80 and 85. An IL-18 haplotype was an independent risk factor of poor Performance IQ. The TNF-308GA genotype was related to individual declines in Verbal IQ, and the IL-10-592 CC genotype was related to better Verbal IQ at the age of 80. Circulating levels of TNF-α, sTNFRs, and IL-6 were negatively correlated with IQ at age 85 and less strongly to IQ at age 80 with activation of the TNF system as the strongest biomarker. In conclusion, SNPs related to high proinflammatory or low anti-inflammatory activity are independent risk factors of reduced cognitive function in octogenarians. Only the IL-18 haplotype was associated with inflammation in the peripheral blood and only with regard to circulating TNF-α.

The Role of Soluble TNF Receptors for TNF-α in Uveitis

To investigate the presence of soluble tumor necrosis factor receptors (sTNF-Rs) and TNF-alpha in the ocular fluids of patients with uveitis and the capacity of sTNF-Rs to affect TNF-alpha production by intraocular T cells. Ocular fluid samples were collected from patients with active and inactive uveitis, as well as from control subjects without uveitis. The sTNF-Rs and TNF-alpha levels were measured by enzyme-linked immunosorbent assay (ELISA). T-cell clones (TCCs) were established from intraocular infiltrating cells, and the TCCs were cocultured with recombinant sTNF-Rs or TNF-alpha. The supernatants were measured by ELISA. The neutralization of sTNF-R production by TCCs was evaluated with an anti-human TNF-R antibody. Significantly higher amounts of sTNF-R1 and -R2 were present in the ocular fluids of patients with active uveitis than in the ocular fluids of those with inactive uveitis and in control subjects. Significantly higher amounts of TNF-alpha were present in the ocular fluids of patients with active uveitis than in those with inactive uveitis. Recombinant sTNF-Rs enhanced TNF-alpha production by TCCs in a dose-dependent manner. Similarly, recombinant TNF-alpha enhanced sTNF-Rs production by the TCCs, and production was neutralized with anti-human TNF-R antibody. sTNF-Rs are present in the ocular fluids of patients with uveitis. Intraocular levels of sTNF-Rs are significantly increased in patients with uveitis, particularly in those with active uveitis. The data suggest that intraocular sTNF-Rs may play a regulatory role in ocular inflammation such as occurs in uveitis.