Higher Tumor Necrosis Factor-alpha Research Articles

Abstract 3428: Association of adipokines and inflammation biomarkers with cancer risk in men

Abstract Using pre-diagnosis fasting plasma samples from 4,040 participants of the Shanghai Men’s Health Study which enrolled 61,464 men ages 40-70 during 2001-2006, we prospectively evaluated the associations of adipokines and inflammation biomarkers with cancer risk. Levels of leptin, resistin, adiponectin, plasminogen activator inhibitor type-1 (PAI-1), interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor-alpha (TNFa), monocyte chemoattractant protein-1 (MCP-1), and insulin were measured by Luminex assays, and C-reactive protein (CRP), measured by ACE Clinical Chemistry System. Biomarker levels were categorized by tertile distribution. The Cox regression model was applied to derive hazard ratios (HR) and 95% confidence intervals (CI) for cancer risk in association with biomarkers with adjustment for potential confounders. Principle component analysis (PCA) was performed to assess associations with multiple biomarkers, and tertile scores for each component were used in analyses. During 15-years of follow-up, 318 cancer cases were identified, including 85 lung, 49 gastric, 17 liver, and 61 colorectal cancer (CRC) cases. Analyses adjusted for age, education and income showed that 3rd vs 1st tertile of TNFa (HR=1.7, 95%CI=1.0-3.0) and adiponectin (HR=4.0, 95%CI=1.1-14.4) was associated with lung and liver cancer risk, respectively. High levels of leptin, insulin, and CRP were each associated with CRC risk: respective HRs were 2.3 (95%CI=1.1-4.7), 2.1 (95%CI=1.0-4.2), and 2.0 (95%CI=1.1-3.9). After further adjustment for known cancer risk factors, TNFa-lung cancer (HR=1.8, 95%CI=1.1-3.1); leptin-CRC (HR= 3.2, 95%CI=1.3-7.7); insulin-CRC (HR=2.1, 95%CI=1.0-4.5); and CRP-CRC (HR=2.0, 95%CI=1.0-4.0) associations remained statistically significant. The adiponectin-lung cancer association lost significance. PCA identified 4 principal components (PC). High PC1 score (main loadings: TNFa, leptin, and insulin) was associated with CRC risk (HR=2.3, 95%CI=1.1-5.0 for 3rd vs 1st tertile ptrend=0.02). High PC2 score (main loadings: adiponectin, IL-8, MCP-1) and PC3 (main loadings: resistin and PAI-1) were associated with liver cancer risk: HR=5.2 (95%CI=1.1-24.3) for the 3rd vs 1st tertile, ptrend=0.03 and HR=4.7 (95%CI=1.0-21.7), for the 3rd vs 1st tertile, ptrend=0.03, respectively. High PC4 score (main loadings: IL-6 and CRP) was associated with lung cancer risk (HR=1.7, 95%CI=1.0-3.0, for the 3rd vs 1st tertile, ptrend=0.04). These associations, except the PC1-CRC association, lost significance after further adjustment of known cancer risk factors. Our study found that high plasma levels of insulin, CRP, and their combination were associated with CRC risk, high resistin and adiponectin levels with liver cancer risk, and high TNFa levels with lung cancer risk. Some biomarker-cancer associations may be explained by their correlations with known cancer risk factors, especially smoking. Citation Format: Douglas DeMoulin, Hui Cai, Qiuyin Cai, Wei Zheng, Xiao-Ou Shu. Association of adipokines and inflammation biomarkers with cancer risk in men [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3428.

Autoinducer-2 promotes the colonization of Lactobacillus rhamnosus GG to improve the intestinal barrier function in a neonatal mouse model of antibiotic-induced intestinal dysbiosis

BackgroundHuman health is seriously threatened by antibiotic-induced intestinal disorders. Herein, we aimed to determine the effects of Autoinducer-2 (AI-2) combined with Lactobacillus rhamnosus GG (LGG) on the intestinal barrier function of antibiotic-induced intestinal dysbiosis neonatal mice.MethodsAn antibiotic-induced intestinal dysbiosis neonatal mouse model was created using antibiotic cocktails, and the model mice were randomized into the control, AI-2, LGG, and LGG + AI-2 groups. Intestinal short-chain fatty acids and AI-2 concentrations were detected by mass spectrometry and chemiluminescence, respectively. The community composition of the gut microbiota was analyzed using 16S rDNA sequencing, and biofilm thickness and bacterial adhesion in the colon were assessed using scanning electron microscopy. Transcriptome RNA sequencing of intestinal tissues was performed, and the mRNA and protein levels of HCAR2 (hydroxycarboxylic acid receptor 2), claudin3, and claudin4 in intestinal tissues were determined using quantitative real-time reverse transcription PCR and western blotting. The levels of inflammatory factors in intestinal tissues were evaluated using enzyme-linked immunosorbent assays (ELISAs). D-ribose, an inhibitor of AI-2, was used to treat Caco-2 cells in vitro.ResultsCompared with the control, AI-2, and LGG groups, the LGG + AI-2 group showed increased levels of intestinal AI-2 and proportions of Firmicutes and Lacticaseibacillus, but a reduced fraction of Proteobacteria. Specifically, the LGG + AI-2 group had considerably more biofilms and LGG on the colon surface than those of other three groups. Meanwhile, the combination of AI-2 and LGG markedly increased the concentration of butyric acid and promoted Hcar2, claudin3 and claudin4 expression levels compared with supplementation with LGG or AI-2 alone. The ELISAs revealed a significantly higher tumor necrosis factor alpha (TNF-α) level in the control group than in the LGG and LGG + AI-2 groups, whereas the interleukin 10 (IL-10) level was significantly higher in the LGG + AI-2 group than in the other three groups. In vitro, D-ribose treatment dramatically suppressed the increased levels of Hcar2, claudin3, and claudin4 in Caco-2 cells induced by AI-2 + LGG.ConclusionsAI-2 promotes the colonization of LGG and biofilm formation to improve intestinal barrier function in an antibiotic-induced intestinal dysbiosis neonatal mouse model.

Hesperidin alleviates inflammation in the metabolic syndrome model

In metabolic syndrome, activated inflammatory signaling pathways trigger the release of proinflammatory cytokines. Nowadays, the use of natural bioactive compounds is trending as an alternative method for the treatment and management of metabolic syndrome. This study aimed to assess the potential effects of hesperidin in the metabolic syndrome model by analyzing the proinflammatory and anti-inflammatory cytokines in serum and liver. Rats were divided into 4 groups: Control (Rats were fed a standard chow diet and water ad libitum), hesperidin [Rats were fed hesperidin supplemented standard chow diet (1%, 10 g/kg feed) and water ad libitum] metabolic syndrome (Rats were fed standard chow diet with 10% fructose-added-drinking-water), and metabolic syndrome + hesperidin (Rats were fed a hesperidin-added standard chow diet (1%, 10 g/kg) with 10% fructose-added-drinking-water). Rats were sacrificed under ketamine/xylazine anesthesia, blood was obtained and liver tissues were removed. Tumor necrosis factor-alpha, interleukin-1 beta, interleukin-6, interleukin-10, and transforming growth factor-beta in the serum and liver were measured by enzyme-linked immunosorbent assay. In the metabolic syndrome group, higher tumor necrosis factor-alpha, interleukin-1 beta, and interleukin-6, but lower serum and liver interleukin-10 and transforming growth factor-beta were found in the serum and liver compared to the control group. In addition, in the metabolic syndrome + hesperidin group lower interleukin-1 beta and interleukin-6 but higher serum interleukin-10 and transforming growth factor-beta were found in the serum and liver compared to the metabolic syndrome groups. Consequently, hesperidin suppressed the serum and liver proinflammatory cytokine response and stimulated the anti-inflammatory cytokine response in the metabolic syndrome rat model.

Immunometabolic Profile Associated with Progressive Damage of the Intestinal Mucosa in Adults Screened for Colorectal Cancer: Association with Diet.

Environmental factors such as diet and lifestyle have been shown to influence the development of some intestinal mucosal lesions that may be precursors of colorectal cancer (CRC). The presence of these alterations seems to be associated with misbalanced immunological parameter levels. However, it is still unclear as to which immunological parameters are altered in each phase of CRC development. In this work, we aimed to study the potential relationships of immunological and metabolic parameters with diet in a CRC-related lesion context. Dietary information was obtained using an annual semi-quantitative food-frequency questionnaire (FFQ) from 93 volunteers classified via colonoscopy examination according to the presence of intestinal polyps or adenocarcinoma. Cytokines, chemokines, and adipokines were determined from serum samples. We observed a reduction in adiponectin according to the damage to the mucosa, accompanied by an increase and decrease in C-X-C motif chemokine ligand 10 (CXCL10) and resistin, respectively, in CRC cases. The presence of aberrant crypt foci (ACF) in the polyp group was associated with higher tumor necrosis factor-alpha (TNF-α) concentrations. Vegetables were directly correlated with adiponectin and resistin levels, while the opposite occurred with red meat. A bioactive compound, soluble pectin, showed a negative association with TNF-α. Future dietary strategies could be developed to modulate specific immunological parameters in the context of CRC.

Ambient Exposure and Bone Homeostasis: Analysis of UK Biobank Data and Experimental Studies in Mice and in Vitro.

Previous evidence has identified exposure to fine ambient particulate matter () as a leading risk factor for adverse health outcomes. However, to date, only a few studies have examined the potential association between long-term exposure to and bone homeostasis. We sought to examine the relationship between long-term exposure and bone health and explore its potential mechanism. This research included both observational and experimental studies. First, based on human data from UK Biobank, linear regression was used to explore the associations between long-term exposure to (i.e., annual average concentration for 2010) and bone mineral density [BMD; i.e., heel BMD () and femur neck and lumbar spine BMD ()], which were measured during 2014-2020. For the experimental animal study, C57BL/6 male mice were assigned to ambient or filtered air for 6 months via a whole-body exposure system. Micro-computed tomography analyses were applied to measure BMD and bone microstructures. Biomarkers for bone turnover and inflammation were examined with histological staining, immunohistochemistry staining, and enzyme-linked immunosorbent assay. We also performed tartrate-resistant acid phosphatase (TRAP) staining and bone resorption assay to determine the effect of exposure on osteoclast activity in vitro. In addition, the potential downstream regulators were assessed by real-time polymerase chain reaction and western blot. We observed that long-term exposure to was significantly associated with lower BMD at different anatomical sites, according to the analysis of UK Biobank data. In experimental study, mice exposed long-term to exhibited excessive osteoclastogenesis, dysregulated osteogenesis, higher tumor necrosis factor-alpha () expression, and shorter femur length than control mice, but they demonstrated no significant differences in femur structure or BMD. In vitro, cells stimulated with conditional medium of macrophages had aberrant osteoclastogenesis and differences in the protein/mRNA expression of members of the pathway, which could be partially rescued by inhibition. Our prospective observational evidence suggested that long-term exposure to is associated with lower BMD and further experimental results demonstrated exposure to could disrupt bone homeostasis, which may be mediated by inflammation-induced osteoclastogenesis. https://doi.org/10.1289/EHP11646.

Cross-sectional study investigating the association between inflammatory biomarkers and neuropathy in adolescents with type 1 diabetes

ObjectivesThe aims of this study were to investigate circulating levels of inflammatory markers in adolescents with type 1 diabetes with and without different types of neuropathies and evaluate the association...

Causal effects of systemic inflammatory regulators on chronic kidney diseases and renal function: a bidirectional Mendelian randomization study.

While targeted systemic inflammatory modulators show promise in preventing chronic kidney disease (CKD) progression, the causal link between specific inflammatory factors and CKD remains uncertain. Using a genome-wide association study of 41 serum cytokines from 8,293 Finnish individuals, we conducted a bidirectional two-sample Mendelian randomization (MR) analysis. In addition, we genetically predicted causal associations between inflammatory factors and 5 phenotypes, including CKD, estimated glomerular filtration rate (eGFR), dialysis, rapid progression of CKD, and rapid decline in eGFR. Inverse variance weighting (IVW) served as the primary MR method, while MR-Egger, weighted median, and MR-pleiotropy residual sum and outlier (MR-PRESSO) were utilized for sensitivity analysis. Cochrane's Q test for heterogeneity. Leave-one-out method ensured stability of MR results, and Bonferroni correction assessed causal relationship strength. Seventeen cytokines were associated with diverse renal outcomes. Among them, after Bonferroni correction test, higher tumor necrosis factor alpha levels were associated with a rapid decrease in eGFR (OR = 1.064, 95% CI 1.028 - 1.103, P = 0.001), higher interleukin-4 levels were associated with an increase in eGFR (β = 0.003, 95% CI 0.001 - 0.005, P = 0.002), and higher growth regulated oncogene alpha (GROα) levels were associated with an increased risk of CKD (OR=1.035, 95% CI 1.012 - 1.058, P = 0.003). In contrast, genetic susceptibility to CKD was associated with an increase in GROa, and a decrease in eGFR may lead to an increase in stem cell factor. We did not find the presence of horizontal pleiotropy during the analysis. We discovered causally related inflammatory factors that contribute to the initiation and progression of CKD at the genetic prediction level.

Cytokine Gene Expression Profiles during HIV and Helminth Coinfection in Underprivileged Peri-Urban South African Adults.

Intestinal helminth parasites are potent stimulators of T helper type 2 (Th2) and regulatory Th3 anti-inflammatory immune responses, while human immunodeficiency virus (HIV) infections are activators of predominantly T helper type 1(Th1) pro-inflammatory responses. Studies investigating the immune profiles of individuals coinfected with helminths and HIV are scarce. Although it is well known that helminths cause a type 2 immune response during the chronic stage of infection that is characterised by Th2 cell differentiation, eosinophil recruitment, and alternative macrophage activation, the immune mechanisms that regulate tissue damage at the time of parasite invasion are poorly understood. The aim of the study was to determine the cytokine gene expression profiles during HIV and helminth coinfection in underprivileged South African adults living in a peri-urban area with poor sanitary conditions and a lack of clean water supply. Study participants (n = 164) were subdivided into uninfected controls, HIV-infected, helminth-infected, and HIV and helminth-coinfected groups. The Kato-Katz and Mini Parasep techniques and Ascaris lumbricoides-specific Immunoglobulin E (IgE) and Immunoglobulin G4 (IgG4) levels were used to detect helminth infections. Participants' HIV status was determined using two HIV1/2 antibody test kits. RNA was isolated from white blood cells for cytokine (Th1-, Th2-, and Th17-related) and transcription factor gene expression profiling using real-time PCR. Multivariate regression data were adjusted for age, gender, BMI, antiretroviral treatment (ART), and nutritional supplement intake. The HIV and helminth-coinfected group had significantly higher tumour necrosis factor alpha (TNF-α) (adjusted β = 0.53, p = 0.036), interleukin 2 (IL-2) (adjusted β = 6.48, p = 0.008), and interleukin 17 (IL-17) (adjusted β = 1.16, p = 0.001) levels and lower GATA binding protein 3 (GATA3) levels (adjusted β = -0.77, p = 0.018) compared to the uninfected controls. No statistical significance was noted for Th2-related cytokines. The coinfected group had higher proinflammatory Th1- and Th17-related cytokine gene expression profiles compared to the uninfected controls. The findings suggest that pro-inflammatory responses are elevated during coinfection, which supports the hypothesis that helminths have a deleterious effect on HIV immune responses.

Tumour Necrosis Factor Alpha Serum Levels and its Gene Polymorphism 850 C/T (rs1799724) as Prognostic Markers of Traumatic Brain Injury Outcome

Introduction: Road traffic accidents have been responsible for the high incidence of head injuries. In a study from India, it was reported that there were 150 Traumatic Brain Injury (TBI) cases per 1,00,000 population, out of which 20 succumbed to death. In the repair process of TBI many chemicals play a role, of which, Tumour Necrosis Factor alpha (TNF-a) is important as TNF-a was shown to possess both neurotoxic and neuroprotective activity. Aim: To evaluate the serum levels of TNF-a and its functional polymorphism in TBI patients and assessing the outcome. Materials and Methods: The prospective cohort observational study was conducted in the Department of Genetics and Molecular Medicine, Kamineni Hospitals, Hyderabad, Telangana, India, from June 2008 to May 2012. It included 126 patients of both sexes with severe and moderate head injuries based on Glasgow Coma Scale (GCS) scores. A 3 mL of plain/Ethylenediaminetetraacetic acid (EDTA) blood sample was drawn from individuals immediately after admission, which was used for serum TNF-a assessment and DNA isolation for TNF-a genotype analysis. The TNF-a levels were measured by Enzyme Linked Immunosorbent Assay (ELISA) using KRISHGEN Biosystems (Catalog No. KB 100-HTNF a) kit. Outcome assessment was done based on Glasgow Outcome Scale (GOS). Statistical analysis was accomplished by Open Epi version 3.01 software. Odd’s Ratio (OR) were used to calculate risk of good/bad outcome with 95% confidence interval and results with a p<0.05 were considered statistically significant. Results: A total of 126 subjects of Indian origin were enrolled in this study. The mean age of the subjects was 36.7±12.97 years. Out of the 126 patients, 46.8% (59) had multiple lesions. The patients with <40 pg/mL (as per the kit specifications) TNF-a levels were 67.50% (85/126), while those with >40 pg/L were 32.50% (41/126). The most common genotype found in the TBI patients was TT with a frequency of 60.3% (76/126), followed by CT with a frequency of 16.6% (21/126) and CC with a frequency of 23% (29/126). When genotype and TNF-a serum levels were studied together, it was seen that low TNF-a level (11-40 pg/mL) was associated with C allele and CC genotype, while those with TT genotype had high TNF-a level (>40 pg/mL) when compared with other genotypes (CC, CT) and this was found to be statistically significant with an OR=3.4064, (95% CI= 1.8426 to 6.2975), p=0.0001. Higher percentage of death and disability was seen in patients with TT genotype while CC genotype showed good recovery at six months with OR=3.6441 (95% CI=1.911 to 6.946), p=0.0001. This suggests that T allele of TNF-a C850T polymorphism has a 3.6 fold higher risk of bad outcome in patients with TBI. Conclusion: The C allele and CC genotype of rs1799724 polymorphism was associated with positive outcome at three to six months. Thus, evaluating TNF-a levels and evaluating the genotype of rs1799724 polymorphism at admission, can be taken as prognostic marker. It can also be used as a target for therapeutic intervention.

Relationship between adiponectin, TNF\u03b1, and SHBG in prepubertal children with obesity

BackgroundSex hormone-binding globulin (SHBG) levels are low in adult subjects with obesity when compared to normal-weight individuals. Obesity is associated with higher tumor necrosis factor alpha (TNFα) plasma levels and lower adiponectin levels. Moreover, we have recently elucidated the molecular mechanisms by which TNFα and adiponectin regulate hepatic SHBG production.AimThe main objective of this study was to assess if the adult associations between TNFα, adiponectin, and SHBG are present in prepubertal children.MethodsWe determined several morphometric and biochemical parameters in normal-weight (n=15) and obese prepubertal (n=51) children, as well as quantified plasma SHBG, TNFα receptor 1 (TNFα-R1), and adiponectin levels.ResultsOur results showed that prepubertal children with obesity had decreased plasma SHBG levels compared to normal-weight controls (67 nmol/L vs 172 nmol/L). Importantly, SHBG plasma levels correlated significantly (P < 0.05) with TNFα (negatively, ßstd= − 0.31) and adiponectin (positively, ßstd= 0.58) suggesting an important role of these two cytokines in determining plasma SHBG levels in prepubertal children.ConclusionsOur results suggest that plasma adiponectin levels may play a more important role than TNFα in influencing plasma SHBG levels in our prepubertal population with obesity.

Neighborhood Characteristics and Cardiovascular Biomarkers in Middle-Aged and Older Adults: the Baltimore Memory Study.

Neighborhood greenness has been linked to better cardiovascular health, but little is known about its association with biomarkers related to cardiovascular risk. Adverse neighborhood conditions, such as disorder and socioeconomic disadvantage, are associated with higher cardiovascular biomarker levels, but these relationships may differ in neighborhoods with more greenness. This study evaluated cross-sectional associations of validated measures of neighborhood greenness, disorder, and socioeconomic disadvantage with cardiovascular biomarkers in middle-aged and older adults living in Baltimore City. The sample included 500 adults, aged 57-79years, enrolled in the Baltimore Memory Study and living in Baltimore City during 2009-2010. Multi-level log-gamma regressions examined associations between the three neighborhood characteristics and seven cardiovascular biomarkers. Models additionally evaluated the effect modification by neighborhood greenness on associations of neighborhood disorder and socioeconomic disadvantage with the biomarkers. Adjusting for covariates and neighborhood greenness, greater neighborhood disorder was associated with higher C-reactive protein (exp β = 1.21, SE = 0.11, p = 0.035) and serum amyloid A (exp β = 1.28, SE = 0.12, p = 0.008), while greater neighborhood socioeconomic disadvantage was associated with higher tumor necrosis factor alpha (exp β = 1.24, SE = 0.12, p = 0.019). Higher neighborhood greenness was associated with lower soluble vascular cell adhesion molecule-1, accounting for disorder (exp β = 0.70, SE = 0.10, p = 0.010) and socioeconomic disadvantage (exp β = 0.73, SE = 0.10, p = 0.025). There was no evidence of effect modification among neighborhood characteristics. The findings suggest that neighborhood effects on cardiovascular health may be mediated through cardiovascular biomarker levels, and that socioeconomic disadvantage, disorder, and greenness may each be important features of neighborhoods.

Establishing a Differential Marker Profile for Pregnancy Complications Near Delivery

Objective: The aim of this work was to define a differential marker profile for pregnancy complications near delivery. Methods: We enrolled pregnant women who were referred to the outpatient pregnancy clinic of the University Medical Center, Ljubljana, Slovenia, due to symptoms of pregnancy complications and women with a history of pregnancy complications attending the high-risk hospital clinic for close surveillance. They were evaluated for prior risk and were tested for biophysical and biochemical markers at the time of enrolment. Biochemical markers included the pro- and anti-angiogenic markers, along with additional previously reported markers of potential value, all tested by various formats of immuno-diagnostics. Biophysical markers included blood pressure, sonographic markers, and EndoPAT. Statistical differences were determined with Kruskal-Wallis and Mann-Whitney tests for continuous parameters, and Pearson χ² for categorical values. p < 0.05 was considered significant. Results: The cohort included 125 pregnant patients, 31 developed preeclampsia (PE) alone (13 were <34 weeks’ gestation), 16 had intrauterine growth restriction (IUGR) alone (12 were <34 weeks), 42 had both IUGR and PE (22 were <34 weeks), and 15 had an iatrogenic preterm delivery (PTD; 6 were <34 weeks). Twenty-one were unaffected and delivered a healthy baby at term. Mean arterial blood pressure and proteinuria were significantly higher in PE and PE+IUGR but not in pure IUGR or PTD. In PE, IUGR, and PE+IUGR, the levels of soluble fms-like tyrosine kinase 1 (sFlt-1) and soluble endoglin (sEng) were significantly higher, while placental growth factor (PlGF) was very low compared to unaffected controls and PTD. PE, IUGR, and PE+IUGR also had a high anti-angiogenic ratio (sFlt-1/PlGF) and a low proangiogenic ratio of PlGF/(sFlt-1+Eng). Levels of inhibin A were significantly higher in pure PE across subgroups but had many extreme values, which made it a poor differentiator. Higher uterine artery Doppler pulsatility indexes were detected in PE, IUGR, and PE+IUGR, with similar resistance indexes and peaks of systolic velocity. A significantly different marker level between PE and IUGR was found using arterial stiffness that was 10 times higher in PE; concurrently with an increase of the reactive hyperemia index, both were accompanied by a slight increase in placental protein 13. Higher tumor necrosis factor alpha (TNFα) differentially identified iatrogenic very early PTD (<34 weeks). Conclusion: Arterial stiffness can serve as a major marker to differentiate PE (with/without IUGR) from pure IUGR near delivery. TNFα can differentiate iatrogenic early PTD from other complications of pregnancy and term IUGR.

S888 TNF‐ALPHA PROMOTES CLONAL EXPANSION OF KIT D816V+ CELLS IN SYSTEMIC MASTOCYTOSIS

Background:Systemic mastocytosis (SM) is characterized by clonal mast cell (MC) infiltration in the bone marrow (BM). Most patients harbor the activating KIT mutation D816V. Cytokines are substantially involved in the interactions between neoplastic cells and the BM microenvironment in SM and in classical myeloproliferative neoplasms (MPN) and have been recognized as a major trigger of disease evolution. The pro‐apoptotic/anti‐proliferative and pro‐inflammatory cytokine tumor necrosis factor alpha (TNFa) has been identified as an important driver for the expansion of JAK2 V617F+ clones and the establishment of MPN.Aims:Investigation of the potential role of TNFa in disease evolution of SM patients.Methods:In vitro and in vivo experiments with MC lines and primary material of SM patients.Results:In a first step, lentivirally‐driven expression of the KIT D816V mutant gene in the human Mo7e cells suggested an oncogene‐dependent regulation of the production of TNFa. Correspondingly, the human neoplastic KIT D816V+ MC lines HMC‐1.2 and ROSAKIT D816V expressed and secreted substantial amounts of TNFa and inhibition of KIT D816V either by the recently approved multi‐kinase inhibitor midostaurin or by RNAi‐mediated knockdown of KIT significantly reduced expression of this cytokine. Mechanistically, we studied auto‐/paracrine effects of TNFa on the clonal selection of neoplastic MC using KIT D816V negative (ROSAKIT WT) and positive (ROSAKIT D816V) subclones of ROSA cells. TNFa reduced proliferation of ROSAKIT WT cells (84.9% ± 4.6% of control, p&lt;0.01) whereas ROSAKIT D816V cells were resistant to TNFa (102.7% ± 3.7% of control, n.s.). Preliminary data also indicate a relative resistance of primary neoplastic cells obtained from KIT D816V+ SM patients against TNFa in colony‐forming unit (CFU) assay. To determine the mechanism of TNFa resistance in KIT D816V+ cells, we screened for key regulators of apoptosis and identified the anti‐apoptotic protein survivin to be highly upregulated in KIT D816V+ ROSA cells upon TNFa treatment but not in KIT D816V‐ ROSA cells. We hypothesize that KIT D816V+ MC produce substantial amounts of TNFa that induces apoptosis and suppresses proliferation of normal BM cells while MC themselves are relatively resistant via upregulation of survivin. In a next step, we studied the effect of TNFa on growth of neoplastic MC in vivo in a xenotransplant model using ROSAKIT D816V cells with and without knockout of TNFa in NOD‐SCID IL‐2Rγ‐null (NSG) mice. CRISPR/Cas9‐mediated knockout completely abolished expression and secretion of TNFa in ROSAKIT D816V cells and resulted in a significantly longer survival of mice compared to mice injected with TNFa expressing controls (median 49 vs. 58 days, p &lt; 0.05). Finally, the clinical relevance of TNFa expression was studied in a SM patient's cohort (n = 45). TNFa levels in sera of patients were significantly higher (median 1.95, range 0.72–16.64 pg/ml, p = 0.002) compared to age‐ and sex‐matched controls (median 1.49, range 0.51–5.63 pg/ml). Importantly, when we stratified SM patients into those with clearly elevated TNFa levels and those with TNFa levels within the reference range of controls (≤3.63 pg/ml) a significantly lower overall survival was observed for SM patients with high TNFa serum levels (median 6.3 years vs. not reached, p &lt; 0.005).Summary/Conclusion:In summary, we have identified TNFa as a critical, KIT D816V‐dependent, cytokine‐mediator that promotes clonal expansion of KIT D816V+ MC and may thereby be involved in clonal evolution and disease progression in SM.

Correlation Between the Frequency of Painful Crises in Sickle Cell Anemia and the Biomarkers of Inflammation, Endothelial Dysfunction, Coagulation Activation and Bone Metabolism

Objective: Sickle cell anemia is a widespread inherited hemolytic anemia, characterized by chronic hemolysis, infections, recurrent occlusion of the microcirculation and painful crises. In this cross-sectional study, we investigated the correlation between biomarkers of inflammation, endothelial dysfunction, coagulation activation and bone metabolism, with the frequency of painful crisis in sickle cell anemia adult patients, to understand the potential role of these biomarkers in the management of sickle cell anemia.&#x0D; Methods: Sixty-three sickle cell anemia patients were enrolled in this study. They were divided into three groups based on the frequency of pain crises to mildly, moderately, and severely affected. Twenty-five healthy volunteers were enrolled as controls. Plasma levels of different biomarkers were tested, including tumor necrosis factor alpha, soluble intercellular adhesion molecule-1, D-dimer, sclerostin, and nitric oxide.&#x0D; Results: Tumor necrosis factor alpha, soluble intercellular adhesion molecule-1, and D-Dimer plasma levels were significantly higher in sickle cell anemia patients when compared to the controls, while nitric oxide levels were notably lower in the patients' group. Sickle cell anemia patients with severe pain showed higher tumor necrosis factor alpha, soluble intercellular adhesion molecule-1 and D-dimer levels than those mildly and moderately affected with painful crisis. Positive correlations were observed between all measured biomarkers and the frequency of the painful crisis, except for nitric oxide, which showed a negative correlation.&#x0D; Conclusion: Tumor necrosis factor alpha, soluble intercellular adhesion molecule-1, nitric oxide, and D-dimer can be used to assess sickle cell anemia severity and prognosis, and thus may affect determining appropriate management strategies.

Systemic recovery and therapeutic effects of transplanted allogenic and xenogenic mesenchymal stromal cells in a rat blunt chest trauma model

BackgroundEffective therapy of Acute Lung Injury (ALI) is still a major scientific and clinical problem. To define novel therapeutic strategies for sequelae of blunt chest trauma (TxT) like ALI/Acute Respiratory Distress Syndrome, we have investigated the immunomodulatory and regenerative effects of a single dose of ex vivo expanded human or rat mesenchymal stromal cells (hMSCs/rMSCs) with or without priming, immediately after the induction of TxT in Wistar rats. MethodsWe analyzed the histological score of lung injury, the cell count of the broncho alveolar lavage fluid (BAL), the change in local and systemic cytokine level and the recovery of the administered cells 24 h and 5 days post trauma. ResultsThe treatment with hMSCs reduced the injury score 24 h after trauma by at least 50% compared with TxT rats without MSCs. In general, TxT rats treated with hMSCs exhibited a lower level of pro-inflammatory cytokines (interleukin [IL]-1B, IL-6) and chemokines (C-X-C motif chemokine ligand 1 [CXCL1], C-C motif chemokine ligand 2 [CCL2]), but a higher tumor necrosis factor alpha induced protein 6 (TNFAIP6) level in the BAL compared with TxT rats after 24 h. Five days after trauma, cytokine levels and the distribution of inflammatory cells were similar to sham rats. In contrast, the treatment with rMSCs did not reveal such therapeutic effects on the injury score and cytokine levels, except for TNFAIP6 level. ConclusionTxT represents a suitable model to study effects of MSCs as an acute treatment strategy after trauma. However, the source of MSCs has to be carefully considered in the design of future studies.

Role of Hypothalamic Pituitary Adrenal (HPA) Axis and Tumor Necrosis Factor (TNF) in Development of Post-Traumatic Nosocomial Pneumonia

Background and purpose: Infection after experimental focal ischemia may result from brain-induced immune depression. A strong cytokine mediated anti-inflammatory response was recently observed in stroke patients at higher risk of infection, although infection due to the decreased pro-inflammatory mediators can be expected as well. To investigate this question the following experiment was performed. Methods: 105 over 60 years hip fracture patients were included in the study. Sera and lympho/monocytes were separated from blood samples taken on different days (day 1, 3, 6, 9). Isolated lympho/ monocytes were cultured for one day with or without endotoxin (LPS). TNFα levels in sera and in the culture supernatants were determined by bioassays using WEHI 164 cells. Plasma ACTH and cortisol values were measured by RIA kits. Results: From 105 hip fracture patients 7 nosocomial pneumonia patients were found, (4 survivors and 3 nonsurvivors) furthermore 2 non-survivors with cardiovascular death without infection. On the day of trauma the level of circulating TNFα activity was extremely low in nosocomial pneumonia patients in comparison to uneventful healing patients. In pneumonia patients TNFα started to increase on day 3, increased till day 9 then reached the values of uneventful healing group. In two patients with later cardiovascular complications, extremely high TNF alpha activities were detected throughout the entire observation period. The plasma cortisol values were high in nosocomial pneumonia patients in comparison to uneventful healing persons, and decreased slightly by the 9th post-trauma day. In the two cardiovascular patients, serum cortisol was extremely low on the day of trauma and increased gradually during the investigation period. ACTH level was stable in the sera of uneventfully healing patients, while showed large individual and also time-dependent fluctuations people with post-traumatic complications. Conclusions: An excessive decrease in pro-inflammatory response is a key facilitating factor for the development of infection.

Tumor necrosis factor alpha polymorphisms are associated with Parkinson's disease age at onset.

The role of neuroinflammation in Parkinson's disease (PD) has been demonstrated through several different approaches. It was suggested an inflammation-derived oxidative stress and cytokine-dependent toxicity role in the nigrostriatal pathway degeneration and hasten progression of disease. Tumor necrosis factor alpha (TNFA) gene promoter polymorphisms might alter the expression of this cytokine contributing to the pro- and anti-inflammatory polarization. An increased TNFA expression might lead to inflammatory profile predominance. The aim of study was to determine if TNFA haplotypes are associated with PD age at onset. Five polymorphisms in TNFA gene were investigated in 226 patients with idiopathic PD in relation to age at onset. Haplotype grouping was based on allele expression. Logistic binary regression analysis showed that the genetic background leading to higher TNF-α expression confers a higher risk to develop PD earlier. Gender and ancestry did not differ between groups. High TNFA expression may contribute for faster dopaminergic neuron degeneration. In this context, a higher genetic pro-inflammatory profile confers a higher risk to develop PD earlier.

Randomized clinical trial of expressive writing on wound healing following bariatric surgery.

Writing emotionally about upsetting life events (expressive writing) has been shown to speed healing of punch-biopsy wounds compared to writing objectively about daily activities. We aimed to investigate whether a presurgical expressive writing intervention could improve surgical wound healing. Seventy-six patients undergoing elective laparoscopic bariatric surgery were randomized either to write emotionally about traumatic life events (expressive writing) or to write objectively about how they spent their time (daily activities writing) for 20 min a day for 3 consecutive days beginning 2 weeks prior to surgery. A wound drain was inserted into a laparoscopic port site and wound fluid analyzed for proinflammatory cytokines collected over 24 hr postoperatively. Expanded polytetrafluoroethylene tubes were inserted into separate laparoscopic port sites during surgery and removed after 14 days. Tubes were analyzed for hydroxyproline deposition (the primary outcome), a major component of collagen and marker of healing. Fifty-four patients completed the study. Patients who wrote about daily activities had significantly more hydroxyproline than did expressive writing patients, t(34) = -2.43, p = .020, 95% confidence interval [-4.61, -0.41], and higher tumor necrosis factor-alpha, t(29) = -2.42, p = .022, 95% confidence interval [-0.42, -0.04]. Perceived stress significantly reduced in both groups after surgery. Expressive writing prior to bariatric surgery was not effective at increasing hydroxyproline at the wound site 14 days after surgery. However, writing about daily activities did predict such an increase. Future research needs to replicate these findings and investigate generalizability to other surgical groups. (PsycINFO Database Record

Slow infusion rate of doxorubicin induces higher pro-inflammatory cytokine production

Different infusion rates of doxorubicin (DOX) have been used for treating human malignancies. Organ toxicity after DOX infusion is a major issue in treatment disruption. However, whether different DOX infusion rates induce different toxicity is still unknown. In this study, we examined the toxicity effects of different DOX infusion rates in the early phase of organ toxicity. Thirty-six rats were randomly divided into 5-, 15-, and 30-min infusion rate groups. A single dose of DOX (8.3 mg/kg, I.V.) was administered at different infusion rates. Blood samples were collected from the femoral artery at 1, 3, 6, 9, 12, 18, 24, 36, and 48 h after DOX administration. The blood cell count and blood biochemistry were analyzed. The liver, kidney, and heart were removed for pathological examinations after the rats were sacrificed. Our findings show that the 30-min group had higher injury markers in the liver (glutamic oxaloacetic transaminase and glutamic pyruvic transaminase), kidneys (blood urea nitrogen and creatinine), and heart (creatine phosphokinase-MB and lactate dehydrogenase), and had higher tumor necrosis factor-alpha and interleukin 6 levels than did the other groups. The 30-min group also had more severe damage according to the pathological examinations. In conclusion, slower infusion of DOX induced a higher inflammatory response and greater organ damage.

A Blunted Response to the Anti-Inflammatory Cytokine IL-10 Contributes to Excessive TNF Production in Myeloproliferative Neoplasm

Rationale:Tumor Necrosis Factor-alpha (TNF) is elevated in myeloproliferative neoplasm (MPN) and plays a key role in expansion of the JAK2V617F neoplastic clone. A high TNF environment, as is the case in MPN patients, gives TNF resistant JAK2V617F mutant cells a selective advantage over their TNF sensitive non-mutant counterparts resulting in expansion of the neoplastic clone. To efficiently target TNF production therapeutically it is necessary to identify the mechanism driving this excessive TNF production. TNF is classically produced by monocytes after stimulation through Toll-like receptors (TLR), crucial pattern recognition receptors for microbial products. Because TLR signaling plays an integral role in inflammation and TNF production, we hypothesized that exaggerated signaling of the TLR pathway is the mechanism by which TNF is overproduced in MPN.Results:We compared the response of peripheral blood monocytes from MPN versus normal controls to the TLR agonists R848 (TLR7/8), LPS (TLR4), or zymosan (TLR2). After stimulation with each of these TLR agonists for 24 hours, CD14+ monocytes from MPN patients (n=18) produced increased amounts of TNF (measured by ELISA) as compared to normal controls (n=10) at all concentrations tested (p<0.05). The increased TNF production in MPN monocytes could not be explained by a higher fraction of inflammatory (CD16+ CD14+) monocytes in MPN. We next used phosflow to detect whether MPN patients have abnormal activation of downstream signaling molecules following stimulation with TLR agonists. At early time points (15min) following stimulation, MPN patients (n=6) and normal controls (n=6) phosphorylated p38 and ERK1/2 equally. At later time points (2hrs) MPN patients maintained phosphorylation of p38 and ERK1/2, whereas in normal controls phosphorylation of p38 and ERK1/2 returned to baseline (p<0.05).This suggested that the negative feedback regulation of TLR signaling may be defective in MPN patients resulting in prolonged TNF production after stimulation. To test this, we compared the tempo of TNF production following LPS stimulation in MPN versus normal monocytes using intracellular flow cytometry as well as ELISA. Monocytes from normal controls (n=12) cease production of TNF by 9 hours post LPS stimulation, whereas MPN monocytes (n=12) continue to produce TNF at 18 hours (p<0.05).LPS stimulation induces a negative feedback loop culminating in the production of IL-10, an anti-inflammatory cytokine which serves to dampen TNF production. We found that MPN monocytes produce abundant IL-10 in response to LPS stimulation. Next we measured the ability of IL-10 to dampen TNF production in LPS stimulated monocytes comparing MPN patients versus normal controls. Addition of low concentrations of rhIL-10 (1ng/ml) to LPS stimulated monocytes reduced TNF production by 50% in normal controls (n=8), whereas it reduced TNF production by only 10-20% in MPN (n=5). However, higher concentrations of IL-10 (>10ng/ml) was able to dampen TNF production in LPS stimulated MPN monocytes by 50%.Conclusions:MPN monocytes produce excessive amounts of TNF in response to TLR ligation due to a defect in the negative regulatory feedback loop which normally serves to dampen TNF production following TLR ligation. We have localized this defect to a blunted response to the anti-inflammatory cytokine IL-10. MPN monocytes are less responsive to the anti-inflammatory actions of IL-10 at low concentrations but these effects can be restored by increasing the concentration of IL-10. Together, these data suggest that administration of IL-10 may reduce excessive inflammatory cytokine production in MPN. DisclosuresNo relevant conflicts of interest to declare.