Plasma Concentrations Of TNF-alpha Research Articles

Investigating the Role of Serum and Plasma IL-6, IL-8, IL-10, TNF-alpha, CRP, and S100B Concentrations in Obstructive Sleep Apnea Diagnosis.

Obstructive sleep apnea (OSA) is a prevalent and underdiagnosed condition associated with cardiovascular diseases, depression, accidents, and stroke. There is an increasing need for alternative diagnostic tools beyond overnight sleep studies that measure the Apnea/Hypopnea Index (AHI). In this single-center, case-control study, we evaluated serum and plasma concentrations of IL-6, IL-8, IL-10, TNF-α, CRP, and S100B in 80 subjects, including 52 OSA patients (27 moderate [15 ≤ AHI ˂ 30], 25 severe [AHI ≥ 30]) and 28 non-OSA controls (AHI 0-5). Participants with OSA showed approximately 2 times higher median concentrations of CRP in plasma, and IL-6 in serum, as well as 1.3 to 1.7 times higher concentrations of TNF-α and IL-8 in plasma compared with the control group. Receiver Operator Characteristic (ROC) curve analysis was performed to evaluate the predictive capabilities of these serum and plasma biomarkers in distinguishing between the OSA and control groups, revealing varying sensitivity and specificity. In summary, in this study, serum and plasma biomarkers CRP, S100B, IL-6, TNF-α, and IL-8 have been shown to be elevated in patients with OSA, correlated positively with disease severity, age, and BMI. These results support the potential role of these biomarkers in diagnosing OSA, supplementing traditional methods such as overnight sleep studies.

MicroRNAs in bovine milk exosomes are bioavailable in humans but do not elicit a robust pro-inflammatory cytokine response

BackgroundBovine milk exosomes are studied for their roles as bioactive food compounds and as vehicles for drug delivery. Both lines of investigation converge on immune function, e.g., immune regulation by absorption of microRNAs encapsulated in milk exosomes across species boundaries, and the possibility of exosomes and their cargos triggering an immune response if used in drug delivery. This study assessed the bioavailability of immune-related microRNAs from bovine milk and changes in plasma cytokine concentrations after milk consumption in humans, and the secretion of cytokines by human peripheral blood mononuclear cells (PBMCs) cultured with milk exosomes transfected with immune-relevant microRNAs.ResultsHuman plasma samples were collected before and at timed intervals after a milk meal and analyzed for concentrations of six immune-relevant microRNAs and nine cytokines. The peak plasma concentrations of miR-15b-5p, miR-21-5p, miR-106b-5p, and miR-223-3p were 60 ± 9.80% to 162 ± 31.80% higher after milk consumption (Ct values 23 ± 1.2 to 26 ± 1.1 cycles) compared to baseline values (P < 0.05). Plasma concentrations of TNF-alpha were not significantly different before versus after milk consumption; eight other cytokines were below detection limit. PBMCs were collected before and six hours after milk consumption and cultured with or without concanavalin A (ConA). TNF-alpha, IL-1β, IL-6 and IL-10 were detectable in culture media, but concentrations did not depend on milk consumption prior to PBMC isolation (P > 0.05). When PBMC cultures from fasted subjects were supplemented with milk exosomes that had been transfected with immune-relevant microRNAs, the concentrations of IL-1β, IL-6, IL-10 and TNF-alpha were 29 ± 12% to 220 ± 33% higher than controls cultured with non-transfected exosomes (P < 0.05), but cytokine concentrations were not different compared with control exosomes transfected with scrambled microRNA (P > 0.05).ConclusionsMicroRNAs in bovine milk exosomes are bioavailable. Milk exosomes do not elicit an increase of plasma cytokines following oral administration.Trial registrationISRCTN registry ID: 16329971. Retrospectively registered on February 7th, 2019.

Abstract #4294 Plasma concentrations of CRP, IL-6, IL-10, and TNF-alpha in patients with depression before and after treatment with electroconvulsive therapy

Abstract #4294 Plasma concentrations of CRP, IL-6, IL-10, and TNF-alpha in patients with depression before and after treatment with electroconvulsive therapy

Correlations between parameters of central hemodynamics and cytokine profile in chronic kidney disease in combination with cerebrovascular diseases

To study the relationship between central hemodynamics and arterial stiffness indicators with cystatin C, tumor necrosis factor-alpha (TNF-alpha) and interleukin-(IL)-10 in patients with chronic kidney disease (CKD) in combination with cerebrovascular diseases (CEV). One hundred and twenty patients, aged from 19 to 81 years, with signs of chronic renal dysfunction, including 73 with CKD (1st group) and 47 with CKD in combination with CEH (group 2), were examined. 'AngioScan' (Russia) was used to study indices of arterial rigidity. Blood plasma concentrations of TNF-alpha, IL-10 and cystatin C were determined by the enzyme immunoassay. There is a significant increase (p<0.05) in the systolic, diastolic, central levels of arterial pressure, augmentation index, the index of increase at a pulse rate of 75 per min, the age index, the age of the vascular system and cystatin C content in the patients with CKD in combination with CEV compared to the patients with CKD without accompanying CEH. In the group of patients with CKD in combination with CEV, a correlation was found between the level of TNF-alpha and arterial stiffness index on one side (r=0.318; p<0.05) and the augmentation index on the other (r=0.299; p<0.05). The results confirm the fact that there is a significant increase in the level of plasma cystatin C and a decrease in GFR in patients with CKD in combination with CEV. Correlations were found between the level of TNF-alpha, augmentation index and deterioration of parameters of central hemodynamics and arterial stiffness in CKD in combination with CEH.

Paternal sepsis induces alterations of the sperm methylome and dampens offspring immune responses\u2014an animal study

BackgroundSepsis represents the utmost severe consequence of infection, involving a dysregulated and self-damaging immune response of the host. While different environmental exposures like chronic stress or malnutrition have been well described to reprogram the germline and subsequently offspring attributes, the intergenerational impact of sepsis as a tremendous immunological stressor has not been examined yet.MethodsPolymicrobial sepsis in 12-week-old male C57BL/6 mice was induced by cecal ligation and puncture (CLP), followed by a mating of the male survivors (or appropriate sham control animals) 6 weeks later with healthy females. Alveolar macrophages of offspring animals were isolated and stimulated with either LPS or Zymosan, and supernatant levels of TNF-α were quantified by ELISA. Furthermore, systemic cytokine response to intraperitoneally injected LPS was assessed after 24 h. Also, morphology, motility, and global DNA methylation of the sepsis survivors’ sperm was examined.ResultsComparative reduced reduction bisulfite sequencing (RRBS) of sperm revealed changes of DNA methylation (n = 381), most pronounced in the intergenic genome as well as within introns of developmentally relevant genes. Offspring of sepsis fathers exhibited a slight decrease in body weight, with a more pronounced weight difference in male animals (CLP vs. sham). Male descendants of sepsis fathers, but not female descendants, exhibited lower plasma concentrations of IL-6, TNF-alpha, and IL-10 24 h after injection of LPS. In line, only alveolar macrophages of male descendants of sepsis fathers produced less TNF-alpha upon Zymosan stimulation compared to sham descendants, while LPS responses kept unchanged.ConclusionWe can prove that male—but surprisingly not female—descendants of post-sepsis fathers show a dampened systemic as well as pulmonary immune response. Based on this observation of an immune hypo-responsivity, we propose that male descendants of sepsis fathers are at risk to develop fungal and bacterial infections and might benefit from therapeutic immune modulation.

CARD19: a mitochondrial caspase recruitment domain protein with a role in regulating pro-inflammatory innate immune responses

Abstract CARD19 is a ubiquitously expressed protein of unknown function. This protein was originally named BinCARD, and early data suggested that it interacts with Bcl10 and inhibits NF-κB activation in T cells. However, these early data were based on overexpression studies, and we are aware of no in vivo data that support this proposed function. Additionally, the cellular localization of CARD19 remains uncharacterized. We sought to address these gaps in knowledge by examining macrophages, thymocytes, and splenocytes from Card19 +/+ and Card19 −/− mice using superresolution microscopy and flow cytometry; we also compared secretion of TNF-alpha, IL-6, and MCP-1 in Card19 +/+ and Card19 −/− mice injected with LPS. Superresolution microscopy revealed that CARD19 is localized to the outer mitochondrial membrane, with no evidence of co-localization with BCL10. Deletion of the predicted transmembrane domain resulted in nuclear and cytosolic distribution, while a partial deletion of the C-terminus resulted in localization in the endoplasmic reticulum. Data to date do not indicate significant differences between Card19 +/+ and Card19 −/− thymocytes and splenocytes in development, activation, or effector function. Interestingly, Card19 −/− mice displayed higher plasma concentrations of TNF-alpha, IL-6, and MCP-1 relative to Card19 +/+ mice in response to LPS. We conclude that CARD19 does not measurably impact BCL10 function in vivo, and we suggest that CARD19 is likely involved in regulation of a mitochondrial function that affects pro-inflammatory innate immune responses via a mechanism independent of BCL10.

P 229 - Ubiquinol supplemenetation in elderly patients undergoing aortic valve replacement: Biochemical and clinical effects

There is a steady rise in the mean age of patients affected by heart disease undergoing cardiac surgery. However senescent myocardium has reduced tolerance to ischemic stress and there are clear indications about age-associated deficit in myocardial performance after operatory stress. CoQ10 improve several conditions related to bioenergetic deficit or increased exposure to oxidative stress. The use of ubiquinol, the reduced form of CoQ10, is promising in these clinical settings, on the basis of its superior bioavailability and of the alleged impaired CoQ10-reducing capacity in the elderly. In this study clinical and biochemical effects of ubiquinol were evaluated in 46 patients undergoing aortic valve replacement. They, affected by severe aortic stenosis, were randomized into 2 groups, placebo and CoQ10 group supplemented with 400 mg/die of ubiquinol starting, 7 days before surgery and continuing for 1 month after cardiac intervention. CoQ10 levels, its oxidative status and IL-6, TNF-alpha and S100 protein concentration in plasma were assessed. Moreover, main cardiac adverse effects in the postoperatory phase, NYHA class, contractility and myocardial hypertrophy were evaluated. Ubiquinol treatment improve basal (before surgery) oxidative status of CoQ10 and considerably mitigate increased oxidation related to the operation.

B- and T-cell subpopulations in patients with severe idiopathic membranous nephropathy may predict an early response to rituximab

Primary membranous nephropathy (PMN) is characterized by antibodies to the podocyte, but little is known about B- and T-cell populations and their response to rituximab is controversial. To help resolve this we compared 33 lymphocyte subpopulations and 27 cytokines/chemokines in 25 patients with severe PMN and 27 age-matched healthy individuals. At baseline, patients had a significantly increased percentage of naive B-cells with significantly decreased switched and non-switched memory B-cells. There was a significantly decreased percentage of natural killer (NK) cells with an increase in the CD56brightCD16-/lo NK subset. There were a significantly decreased percentage of regulatory T cells, together with an increased plasma concentration of TNF-alpha, IL-5 and IL-2RA. We then investigated 16 patients at eight days and three and six months after treatment with rituximab added to supportive therapy compared to nine patients with supportive therapy alone. After rituximab, B-cell recovery was still incomplete at six months, with persistent alterations of B-cell subsets, a significant increase of both T-regulatory (Treg) cells and NK cells, and a significant decrease of both the CD56brightCD16-/lo NK subset and TNF-alpha levels. The patients who clinically responded to rituximab had a significantly lower percentage of Tregs at baseline compared to non-responders and a significantly increased percentage at day eight. Tregs remained unchanged in non-responders and in patients treated with supportive therapy alone. Thus, evaluation of Tregs might be useful for predicting early response to rituximab.

Factor de necrosis tumoral alfa plasmático en mujeres obesas y no obesas con síndrome de ovarios poliquísticos

Objetivo: Determinar las concentraciones plasmáticas de factor de necrosis tumoral alfa (FNT-alfa) en mujeres obesas y no obesas con diagnóstico de síndrome de ovarios poliquísticos (SOPQ). Diseño: Estudio de casos y controles. Participantes: Mujeres con diagnóstico de SOPQ y controles sanas de edades similares, con menstruaciones regulares y ovarios normales por ecografía. Métodos: Las participantes fueron divididas en cuatro grupos (grupo A: SOPQ obesas; grupo B: SOPQ no obesas; grupo C: controles obesas; y grupo D: controles no obesas), de acuerdo al índice de masa corporal (obesas &gt; 30 kg/m2 y no obesas &lt; 25 kg/m2). Principales medidas de resultados: Concentraciones de lutoprina, folitropina, androstendiona, testosterona, globulina fijadora de hormonas sexuales, glucosa sérica, insulina y FNT-alfa. Resultados: Las mujeres con SOPQ obesas y no obesas presentaron concentraciones más elevadas de hormonas sexuales e insulina comparado con el grupo control de obesas y no obesas respectivamente (p &lt; 0,0001). Se observó que las mujeres con SOPQ tuvieron concentraciones significativamente más altas de FNT-alfa (grupo A: 6,6 +/- 1,2 pg/mL y grupo B: 4,0 +/- 0,7 pg/mL) comparado con los controles (grupo C: 4,4 +/- 1,3 pg/mL) y grupo D (2,1 +/- 0,4 pg/dL; p &lt; 0,0001). Se observó que las concentraciones de FNT-alfa presentaban correlación positiva y significativa con los valores de glicemia e insulina en ayunas en las mujeres con SOPQ (p &lt; 0,0001). Conclusión: Se ha hallado diferencias significativas en las concentraciones plasmáticas del FNT-alfa entre las mujeres con SOPQ obesas y no obesas respecto a los controles normales.

Abstract # 1796 Impaired beta-adrenergic upregulation of anti-inflammatory cytokine production in patients with ulcerative colitis in remission

Psychological factors, especially stress, demonstrably contribute to disease course in patients with inflammatory bowel diseases, but the underlying mechanisms remain unclear. Here, we tested whether neuroendocrine regulation of pro-/anti-inflammatory cytokine production by peripheral blood cells is altered in patients with ulcerative colitis in remission (UCR). Using a diluted-whole-blood stimulation assay, we measured the sensitivity of lipopolysaccharide-induced TNF-alpha and IL-10 production to dexamethasone (DEX; glucocorticoid agonist), terbutaline (TERB; beta2-adrenergic agonist), and GTS-21 (alpha7-nicotinic acetylcholine receptor agonist) in samples from UCR patients ( n = 26) and healthy controls ( n = 25). Additionally, we assessed anxiety and depression symptoms as well as chronic perceived stress and health-related quality-of-life with validated questionnaires (HADS, PSQ, IBDQ). Results showed that UCR patients exhibited greater anxiety, depression and chronic stress levels, and reduced quality-of-life. Plasma concentration of TNF-alpha was significantly higher and LPS-induced IL-10 production was significantly lower in UCR compared to HC. Independent of group, DEX and GTS-21 dose-dependently inhibited TNF-alpha and IL-10 production, whereas TERB inhibited TNF-alpha and upregulated IL-10 production. However, at higher TERB doses (i.e., stress levels), upregulation of IL-10 production was significantly diminished in UCR compared to HC, and impaired beta2-adrenergic IL-10 regulation correlated positively with the IBDQ systemic symptom score. These findings extend evidence of peripheral immune alterations in UC even during phases of remission, and support dysregulation of anti-inflammatory mechanisms that could play a role in stress-induced symptom exacerbation.

Abstract # 1713 Predictors of bodily sickness symptoms during experimental endotoxemia

Patients with inflammatory conditions commonly suffer from non-specific bodily symptoms such as fatigue, widespread pain, and malaise. However, it remains unclear if these symptoms are driven by cytokines and/or psychological factors. Experimental administration of endotoxin (LPS) is an established model to induce a wide spectrum of behavioral and physical sickness symptoms. The aim of the present study was to identify predictors of LPS-induced physical sickness symptoms using multivariate regression analyses. Healthy volunteers (N = 128) were randomized to LPS (0.4 or 0.8 ng/kg) or placebo in published studies with identical protocols. Psychological state (state anxiety, positive mood) and trait variables (depression symptoms, trait anxiety, neuroticism, extraversion, coping skills) were measured along with plasma concentrations of IL-6, TNF-alpha, and cortisol. Non-specific sickness symptoms experienced during the acute (0–6 h post-injection) and late inflammatory (6–24 h post-injection) phases were assessed with the validated Generic Assessment of Side Effects (GASE). Results showed that LPS-treated subjects reported significantly more physical sickness symptoms. Symptoms during the early phase were predicted by decreases in positive mood and increases in IL-6 plasma levels, explaining 28.5% of variance in GASE scores. During the late phase, increases in cortisol, IL-6 and baseline depression were significant predictors, explaining 38.5% of variance. Hence, mood alterations may constitute a risk factor for physical sickness symptoms in inflammatory conditions.

Antimicrobial and immunomodulatory effects of tedizolid against methicillin-resistant Staphylococcus aureus in a murine model of hematogenous pulmonary infection

Tedizolid (TZD) is a second-generation oxazolidinone and demonstrates potent in-vitro activity against multidrug-resistant Gram-positive bacteria. Phase III studies in patients with acute bacterial skin and skin structure infections (ABSSSI) have demonstrated the non-inferiority of TZD to linezolid (LZD). However, there are only a few studies that show the effect of TZD in pulmonary infections. In this study, we investigated the effect of TZD in a murine model of hematogenous pulmonary infection caused by methicillin-resistant Staphylococcus aureus (MRSA). The mice were treated either twice daily with saline (control), 25mg/kg of vancomycin (low-VAN), 110mg/kg of vancomycin (high-VAN), 120mg/kg of LZD or once daily with 20mg/kg of TZD. As compared to the control, the low- and high-VAN treatment groups, LZD and TZD significantly improved the survival rate, reduced the bacterial count in the lungs. Furthermore, TZD decreased the area of central bacterial colony zone (CBCZ) at 36h post-inoculation, compared with the control. In addition, we investigated the immunomodulatory effect of TZD by evaluating the plasma concentrations of the inflammatory cytokines. Although there were no significant differences in the bacterial count in the lungs amongst the drugs at 26h post-inoculation, TZD and LZD significantly improved the plasma concentrations of TNF-alpha, IL-6 and MIP-2, in comparison with the control. In this study, both TZD and LZD demonstrated antimicrobial and immunomodulatory efficacy in a murine model of hematogenous pulmonary infection caused by MRSA.

Rapidly Escalating Hepcidin and Associated Serum Iron Starvation Are Features of the Acute Response to Typhoid Infection in Humans.

BackgroundIron is a key pathogenic determinant of many infectious diseases. Hepcidin, the hormone responsible for governing systemic iron homeostasis, is widely hypothesized to represent a key component of nutritional immunity through regulating the accessibility of iron to invading microorganisms during infection. However, the deployment of hepcidin in human bacterial infections remains poorly characterized. Typhoid fever is a globally significant, human-restricted bacterial infection, but understanding of its pathogenesis, especially during the critical early phases, likewise is poorly understood. Here, we investigate alterations in hepcidin and iron/inflammatory indices following experimental human typhoid challenge.Methodology/Principal FindingsFifty study participants were challenged with Salmonella enterica serovar Typhi and monitored for evidence of typhoid fever. Serum hepcidin, ferritin, serum iron parameters, C-reactive protein (CRP), and plasma IL-6 and TNF-alpha concentrations were measured during the 14 days following challenge. We found that hepcidin concentrations were markedly higher during acute typhoid infection than at baseline. Hepcidin elevations mirrored the kinetics of fever, and were accompanied by profound hypoferremia, increased CRP and ferritin, despite only modest elevations in IL-6 and TNF-alpha in some individuals. During inflammation, the extent of hepcidin upregulation associated with the degree of hypoferremia.Conclusions/SignificanceWe demonstrate that strong hepcidin upregulation and hypoferremia, coincident with fever and systemic inflammation, are hallmarks of the early innate response to acute typhoid infection. We hypothesize that hepcidin-mediated iron redistribution into macrophages may contribute to S. Typhi pathogenesis by increasing iron availability for macrophage-tropic bacteria, and that targeting macrophage iron retention may represent a strategy for limiting infections with macrophage-tropic pathogens such as S. Typhi.

Pre-existent anxiety symptoms is associated with modified behavioral response during endotoxemia

Experimental endotoxin administration is a well-established model to analyze the effect of inflammation on the development of mood impairments. However, it remains unknown from previous studies in comparatively small samples whether pre-existing inter-individual differences modulate the behavioral response during endotoxemia. We addressed this question in the present study using a merged database combining data from multiple studies performed at two study sites. In 286 healthy volunteers who either received low-dose intravenous injection of lipopolysaccharide (LPS, 0.4–0.8 ng/kg, n = 168) or saline ( n = 118), plasma concentrations of TNF-alpha and IL-6 were analyzed before and 2, 3.5 and 6 hours post injection together with state anxiety symptoms (STAI). Pre-existent symptoms of anxiety and depression (HADS) were assessed before injection. LPS administration induced an increase in state anxiety 2 hours post injection that was positively correlated with increases in TNF-alpha and IL-6, but not with pre-existent HADS scores (when adjusted for age, sex, BMI, LPS dose, and study design). However, higher levels of pre-existent HADS anxiety symptoms predicted a stronger subsequent reduction in state anxiety at 3.5 and 6 hours post LPS injection. Taken together, these data suggest that although inter-individual differences in anxiety or depression symptoms do not appear to explain the extent of mood impairments during endotoxemia, at least in non-patient samples, they still lead to a modified mood response with a faster and stronger decline.

52. Dose-dependent effects of lipopolysaccharide on memory and social cognition in humans

Clinical and experimental studies demonstrate that inflammation and increased concentrations of pro-inflammatory cytokines are associated with depression-like symptoms and neuropsychological impairments in humans. However, it has not been satisfactorily elucidated to what extent cognitive functions such as memory, attention or social cognition are affected during systemic immune activation. Thus, in a double-blind, randomized, placebo-controlled crossover study, healthy male subjects received intravenous injections of either lipopolysaccharide (low-dose LPS: 0.4 ng/kg, n = 18; high-dose LPS: 0.8 ng/kg, n = 16) or placebo (0.9% saline) on two different study days. Memory (n-back; visual long-term memory task) and social cognition performance (MASC; Reading-the-Mind-in-the-Eyes-Test) were assessed 2–4 h after LPS or saline injection. LPS administration dose-dependently elevated body temperature and plasma concentrations of TNF-alpha, IL-6, IL-10, and IL-1ra and increased negative mood and state anxiety. However, endotoxin administration did not affect accuracy in a working memory paradigm but improved reaction time after high-dose LPS administration. In contrast, long-term memory performance was impaired selectively for emotional stimuli after administration of low-dose LPS only. Finally, there was a significant, albeit relatively small, effect on social cognition after injection of the high-dose LPS. Taken together, this study demonstrates that acute systemic inflammation mildly affects but not necessarily impairs cognitive functions in humans.

Acute effects of intracranial hypertension and ARDS on pulmonary and neuronal damage: a randomized experimental study in pigs

PurposeTo determine reciprocal and synergistic effects of acute intracranial hypertension and ARDS on neuronal and pulmonary damage and to define possible mechanisms.MethodsTwenty-eight mechanically ventilated pigs were randomized to four groups of seven each: control; acute intracranial hypertension (AICH); acute respiratory distress syndrome (ARDS); acute respiratory distress syndrome in combination with acute intracranial hypertension (ARDS + AICH). AICH was induced with an intracranial balloon catheter and the inflation volume was adjusted to keep intracranial pressure (ICP) at 30–40 cmH2O. ARDS was induced by oleic acid infusion. Respiratory function, hemodynamics, extravascular lung water index (ELWI), lung and brain computed tomography (CT) scans, as well as inflammatory mediators, S100B, and neuronal serum enolase (NSE) were measured over a 4-h period. Lung and brain tissue were collected and examined at the end of the experiment.ResultsIn both healthy and injured lungs, AICH caused increases in NSE and TNF-alpha plasma concentrations, extravascular lung water, and lung density in CT, the extent of poorly aerated (dystelectatic) and atelectatic lung regions, and an increase in the brain tissue water content. ARDS and AICH in combination induced damage in the hippocampus and decreased density in brain CT.ConclusionsAICH induces lung injury and also exacerbates pre-existing damage. Increased extravascular lung water is an early marker. ARDS has a detrimental effect on the brain and acts synergistically with intracranial hypertension to cause histological hippocampal damage.Electronic supplementary materialThe online version of this article (doi:10.1007/s00134-011-2232-2) contains supplementary material, which is available to authorized users.

Abstract 4646: Occupational exposure to trichloroethylene and plasma concentrations of IL6, IL10, and TNF alpha

Abstract Trichloroethylene (TCE) is a chlorinated solvent primarily used as a vapor degreaser in many industries. TCE is currently classified by IARC as probably carcinogenic to humans (Group 2A), and has been associated with non-Hodgkin lymphoma (NHL) in some epidemiological studies, although the findings have been inconsistent. To explore the biologic plausibility that TCE may be a lymphomagen, we conducted a cross-sectional study of workers exposed to TCE in Guangzhou, China and tested if plasma levels of the cytokines IL6, IL10, and TNF-alpha, which play a critical role in regulating various components of the immune system, were altered in exposed workers. We enrolled 80 workers from factories where TCE was used as well as 96 unexposed controls matched by sex and age from food and clothes manufacturing factories. Personal exposure measurements were taken over a three-week period, two to three per subject, in the exposed subjects for a full work shift using a 3M badge, as well as for a subgroup of the control workers. Blood samples were collected from each subject, and plasma concentrations of IL6, IL10, and TNF-alpha were measured by ELISA. Linear regression using the natural logarithm of each endpoint and adjusting for potential confounders was used to test for differences in marker concentrations between controls and exposed workers. The mean TCE exposure level in exposed subjects was 23.9 ppm (95% C.I. 14.2-33.6 ppm). Compared to unexposed workers, the plasma concentration of IL10 in workers exposed to TCE was decreased by more than 50% compared to controls (p=0.003), while there was no difference in levels of IL6 or TNF-alpha in TCE exposed workers and controls. IL10 plays a critical role in mediating the Th1/Th2 balance, apoptosis, and regulation of inflammation. Genotypes associated with altered IL10 expression have also been associated with the development and progression of NHL. Given that immunologic alterations are suspected to play a role in lymphomagenesis, and IL10 plays an important role in immunologic processes, our findings provide some support for the biologic plausibility that TCE is associated with lymphoma. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4646. doi:10.1158/1538-7445.AM2011-4646

Jak2 V617F Induced TNF Resistance as a Mechanism of Clonal Expansion In Myeloproliferative Neoplasm

AbstractAbstract 795Elevated levels of pro-inflammatory cytokines such as TNF-alpha (TNF) have been implicated in the constitutional symptoms commonly seen in JAK2 V617F MPN. However, it is unclear to what extent they contribute to the disease process itself. We hypothesized that JAK2 V617F may confer a selective advantage on hematopoietic progenitors residing in the high cytokine milieu uniquely present MPN patients, focusing on the pro-inflammatory cytokine TNF.We used ELISA and qPCR to measure TNF in plasma and neutrophils of MPN patients and normal controls. All MPN subtypes had increased plasma concentrations of TNF, with primary myelofibrosis, polycythemia vera and essential thrombocythemia displaying 6, 4 and 2 fold higher mean concentrations of plasma TNF as compared to normal controls, respectively. Median neutrophil TNF mRNA was 2 fold higher in PV patients as compared to normal controls (p<0.001). We then tested the effect of TNF on CFU-GM and BFU-E formation from MPN vs normal mononuclear cells. At low concentrations (1ng/ml) of TNF we observed a decrease in BFU-E from normal controls to 82% of that without TNF but an increase in MPN BFU-E to 120% (p<0.05) of that without TNF. We also observed a blunting of TNF induced CFU-GM suppression in MPN versus normal. We genotyped colonies and found that in the presence of TNF colony formation was almost exclusively JAK2 V617F, demonstrating that TNF selects for JAK2 V617F colony growth. To distinguish between direct and indirect TNF effects (i.e. effects mediated through bystander cells) as well as to potentially augment the differential effects of TNF on normal and MPN by the use of a more homogenous cell population we performed equivalent colony formation experiments on CD34+ cells. The most notable difference observed was the effect 10ng/ml TNF on CFU-GM. TNF reduced CFU-GM colony formation by normal CD34+ cells to 50%, but increased CFU-GM colony formation by MPN cells to 120% (p<0.001). The suppressive effect of 10ng/ml on BFU-E was also blunted in MPN versus normal CD34+ cells (49% vs 30% of that without TNF) (p=0.054). Genotyping of MPN colonies allowed us to compare the effect of TNF on colony formation on three distinct populations: (1) JAK2WT MPN CD34+ cells, (2) JAK2V617F MPN CD34+ cells and (3) normal CD34+ cells. As compared to normal controls (59% decrease), JAK2 non-mutated MPN CD34+ cells are hypersensitive to myeloid colony suppression (78% decrease) whereas TNF paradoxically enhances colony formation by JAK2 V617F CD34+ cells (180% increase). This suggests that “pre-JAK2” CD34+ cells are functionally disadvantaged when compared to those from normal controls and implicates TNF in the selection and clonal expansion of the JAK2 V617F clone. To confirm the latter we tested whether ectopic expression of JAK2 V617F in murine hematopoietic progenitors blunts TNF-induced inhibition of myeloid colony formation. In the presence of TNF, colony formation of hematopoietic progenitors expressing empty vector or BCR-ABL (included as an additional control) was decreased to 60%, while TNF had no effect on progenitors with ectopic expression of JAK2 V617F. This suggests that JAK2 V617F directly imparts TNF resistance. To interrogate the role of TNF for the development of MPN in vivo, we infected TNF+/+ and TNF−/− bone marrow with JAK2 V617F-GFP retrovirus and transplanted the cells into genotypically matched recipients. Two strains of mice were used in independent experiments (C57BL6 and B6129S6). Equal numbers of TNF+/+ and TNF−/− GFP+ cells per mouse were injected. Although peripheral blood counts were comparable, JAK2 V617F burden (assessed by GFP%) was significantly lower in recipients of TNF−/− knockout bone marrow compared to recipients of TNF+/+ marrow. This difference in JAK2 V617F burden between the two groups was apparent in peripheral blood (47% vs 3.7%, p <0.05) and bone marrow (52% vs 2%, p<0.05) and the spleens of the TNF−/− were smaller (0.46g vs 0.11g, p=0.078).Our data suggest that JAK2 V617F-induced TNF resistance may drive the expansion of the mutant clone by conferring a growth advantage to JAK2 V617F cells over their JAK2 wild-type counterparts in the high cytokine microenvironment of MPN patients. Further, JAK2WT MPN cells exhibit intrinsic TNF hypersensitivity compared to normal cells, which greatly enhances this selective advantage. This JAK2V617F-negative state may play a role in the predisposition to acquire MPN. Disclosures:No relevant conflicts of interest to declare.

High habitual dietary α-linolenic acid intake is associated with decreased plasma soluble interleukin-6 receptor concentrations in male twins

alpha-Linolenic acid (ALA) is associated with a low risk of cardiovascular disease; however, the underlying mechanism is not completely known. The objective was to examine whether habitual dietary ALA intake is associated with plasma concentrations of inflammatory biomarkers after control for shared genetic and common environmental factors. We cross-sectionally studied 353 middle-aged male twins. Habitual diet was assessed with the Willett food-frequency questionnaire. Fasting plasma concentrations of interleukin-6 (IL-6) and its soluble receptor (sIL-6R), high-sensitivity C-reactive protein (hsCRP), and tumor necrosis factor-alpha (TNF-alpha) were measured. Linear mixed-effect regression analysis was used to partition the overall association into within- and between-pair associations. A 1-g increment in habitual dietary ALA intake was associated with 11.0% lower concentrations of sIL-6R (P = 0.004) but not of IL-6 (P = 0.31), TNF-alpha (P = 0.16), or hsCRP (P = 0.36) after adjustment for energy intake, nutritional factors, known cardiovascular disease risk factors, and medications. After further control for shared genetic and common environmental factors by comparison of brothers within a twin pair, a twin with a 1-g higher ALA intake was likely to have 10.9% (95% CI: 3.7%, 17.6%; P = 0.004) lower sIL-6R concentrations than his co-twin with a low intake, whereas ALA intake was not significantly associated with plasma concentrations of IL-6, TNF-alpha, or hsCRP. These results were validated by using 1000 bootstrap samples. Habitual dietary ALA intake is inversely associated with plasma sIL-6R concentrations independent of shared genetic and common environmental influences. Lowering sIL-6R may be a mechanism underlying the cardioprotective properties of habitual dietary ALA. This study was registered at clinicaltrials.gov as NCT00017836.

Sleep measures and morning plasma TNF-alpha levels in children with sleep-disordered breathing.

Sleep disordered breathing in children is associated with severity-dependent increases in excessive daytime sleepiness (EDS). TNF-alpha is an inflammatory cytokine that has been implicated in EDS. Since, at any given level of apnea-hypopnea index, there is significant variability in EDS, we hypothesized that morning tumor necrosis factor (TNF)-alpha plasma levels may provide a biologic correlate of EDS. Children being evaluated for sleep disordered breathing underwent a blood draw after nocturnal polysomnography, and TNF-alpha plasma concentrations were assayed using ELISA. In a subset of 15 children with sleep disordered breathing and in 15 matched control subjects, whole blood cultures in the presence of lipopolysaccharide and Multiple Sleep Latency Test were conducted. Furthermore, 22 children with obstructive sleep apnea had TNF-alpha levels assayed and underwent nocturnal polysomnography and Multiple Sleep Latency Test before and after adenotonsillectomy. In 298 children, morning TNF-alpha levels were globally increased in the presence of obstructive sleep apnea, particularly in more severe cases, and correlated with obstructive apnea-hypopnea index and sleep pressure score, a measure of respiratory-induced sleep fragmentation, but not with nadir Sa02. A stepwise logistic regression analysis revealed that sleep pressure score and body mass index accounted for 36.2% of the adjusted variance in TNF-alpha levels (P < 0.0001). Furthermore, multiple sleep latencies were correlated with whole blood culture-derived TNF-alpha levels (n = 15), and morning TNF-alpha levels decreased after adenotonsillectomy in 22 children. TNF-alpha levels are increased in pediatric obstructive sleep apnea, are primarily driven by sleep fragmentation and body mass index, and are closely associated with the degree of sleepiness, as measured by Multiple Sleep Latency Test. Furthermore, surgical treatment of obstructive sleep apnea results in significant reductions in TNF-alpha levels with reciprocal prolongations in sleep latency.