Higher Concentrations Of TNF-α Research Articles

The Effects of IGF-1 on TNF-α-Treated DRG Neurons by Modulating ATF3 and GAP-43 Expression via PI3K/Akt/S6K Signaling Pathway.

Upregulation of the pro-inflammatory cytokine tumor necrosis factor α (TNF-α) is involved in the development and progression of numerous neurological disorders. Recent reports have challenged the concept that TNF-α exhibits only deleterious effects of pro-inflammatory destruction, and have raised the awareness that it may play a beneficial role in neuronal growth and function in particular conditions, which prompts us to further investigate the role of this cytokine. Insulin-like growth factor-1 (IGF-1) is a cytokine possessing powerful neuroprotective effects in promoting neuronal survival, neuronal differentiation, neurite elongation, and neurite regeneration. The association of IGF-1 with TNF-α and the biological effects, produced by interaction of IGF-1 and TNF-α, on neuronal outgrowth status of primary sensory neurons are still to be clarified. In the present study, using an in vitro model of primary cultured rat dorsal root ganglion (DRG) neurons, we demonstrated that TNF-α challenge at different concentrations elicited diverse biological effects. Higher concentration of TNF-α (10ng/mL) dampened neurite outgrowth, induced activating transcription factor 3 (ATF3) expression, reduced growth-associated protein 43 (GAP-43) expression, and promoted GAP-43 and ATF3 coexpression, which could be reversed by IGF-1 treatment; while lower concentration of TNF-α (1ng/mL) promoted neurite sprouting, decreased ATF3 expression, increased GAP-43 expression, and inhibited GAP-43 and ATF3 coexpression, which could be potentiated by IGF-1 supplement. Moreover, IGF-1 administration restored the activation of Akt and p70 S6 kinase (S6K) suppressed by higher concentration of TNF-α (10ng/mL) challenge. In contrast, lower concentration of TNF-α (1ng/mL) had no significant effect on Akt or S6K activation, and IGF-1 administration activated these two kinases. The effects of IGF-1 were abrogated by phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002. These data imply that IGF-1 counteracts the toxic effect of higher concentration of TNF-α, while potentiates the growth-promoting effect of lower concentration of TNF-α, with the node for TNF-α and IGF-1 interaction being the PI3K/Akt/S6K signaling pathway. This study is helpful for interpretation of the association of IGF-1 with TNF-α and the neurobiological effects elicited by interaction of IGF-1 and TNF-α in neurological disorders.

IMMUNOLOGICAL MARKERS OF UNCONTROLLED ATOPIC BRONCHIAL ASTHMA IN CHILDREN

Bronchial asthma is a prevalent chronic allergic disease of lungs at early ages. A priority task in allergology is to search biological markers related to uncontrolled atopic bronchial asthma. Cytokines fulfill their distinct function in pathogenesis of atopic bronchial asthma, participating at the initiation, development and persistence of allergic inflammation in airways, causing different variations of clinical course of the disease (with respect to its acuteness, severity, frequency of exacerbations). The present work has studied indices of cellular and humoral links of immunity, as well as levels of some pro and anti-inflammatory cytokines in peripheral blood serum (IL-4, IL-10, IL-2 and TNFα), aiming to determine potential markers of uncontrolled atopic bronchial asthma in children. A group of Caucasian (European) children was involved into the research: Cohort 1, moderate atopic bronchial asthma with controlled course during the last 3 months (n = 59); Cohort 2, severe/moderate-severe atopic bronchial asthma with uncontrolled course of the disease within last 3 months (n = 51), Cohort 3 – control, practically healthy children without signs of atopy (n = 33). All the children included in the group with atopic bronchial asthma underwent regular mono/combined basic therapy at high/ intermediate therapeutic doses. We performed a comparative analysis of cell population indices reflecting certain cellular immunity links, and determined significantly lower levels of CD3 + lymphocytes, as well as decrease in relative and absolute contents of CD4 + and CD8+ cells in the cohort with uncontrolled course of atopic bronchial asthma, as compared with controlled-course cohort. When evaluating concentrations of cytokines in peripheral blood serum of the patients with controlled and uncontrolled atopic bronchial asthma, we revealed significantly higher levels of IL-2, IL-4, and IL-10, as compared to control group. It was found that TNFα concentration is considerably higher in both cohorts of the patients, being 2to 3-fold higher than the levels of this cytokine in control group. When comparing the cohorts with different control of the disease course, we have found that TNFα concentration in the cohort with uncontrolled bronchial asthma is statistically higher than among children with controllable course of the disease. Hence, the following parameters may serve as potential markers of pediatric atopic bronchial asthma with uncontrolled course: low levels of total Т lymphocyte numbers in peripheral blood, and decreased counts of CD4 + , CD8 + cells; IgE hyperproduction; low contents of common IgA, and high concentration of TNFα in peripheral blood serum.

Pro-Inflammatory Cytokine TNF-α Attenuates BMP9-Induced Osteo/ Odontoblastic Differentiation of the Stem Cells of Dental Apical Papilla (SCAPs)

Background/Aims: Periapical periodontitis is a common oral disease caused by bacterial invasion of the tooth pulp, which usually leads to local release of pro-inflammatory cytokines and osteolytic lesion. This study is intended to examine the effect of TNF-α on BMP9-induced osteogenic differentiation of the stem cells of dental apical papilla (SCAPs). Methods: Rat model of periapical periodontitis was established. TNF-α expression was assessed. Osteogenic markers and ectopic bone formation in iSCAPs were analyzed upon BMP9 and TNF-α treatment. Results: Periapical periodontitis was successfully established in rat immature permanent teeth with periapical lesions, in which TNF-α was shown to release during the inflammatory phase. BMP9-induced alkaline phosphatase activity, the expression of osteocalcin and osteopontin, and matrix mineralization in iSCAPs were inhibited by TNF-α in a dose-dependent fashion, although increased AdBMP9 partially overcame TNF-α inhibition. Furthermore, high concentration of TNF-α effectively inhibited BMP9-induced ectopic bone formation in vivo. Conclusion: TNF-α plays an important role in periapical bone defect during the inflammatory phase and inhibits BMP9-induced osteoblastic differentiation of iSCAPs, which can be partially reversed by high levels of BMP9. Therefore, BMP9 may be further explored as a potent osteogenic factor to improve osteo/odontogenic differentiation in tooth regeneration in chronic inflammation conditions.

TNF-Alpha Promotes Clonal Expansion of Tet2 Deficient Bone Marrow Progenitors in MDS

Background: Recent studies suggest that pro-inflammatory cytokines such as TNF-α are increased in myelodysplastic syndromes (MDS), but their contribution to disease pathogenesis is unknown. Here we uncover a mechanistic role of TNF-α in promoting clonal dominance of Tet2 murine mutant haematopoietic stem and progenitor cells (HSPCs). We hypothesized that Tet2 mutation induces TNFα expression and confers TNFα resistance to MDS progenitor cells by mechanisms involving the suppression of apoptotic cues. Methods: We isolated lineage negative cells (Lin-), enriched for HSPCs, from the bone marrow(BM) of 10-14 weeks old Tet2wild(+/+) type and mutant (-/-) C57BL/6 mice strain, (EasySep; StemCell Technologies) and cultured these +/- TNFα (0.1, 1, or 10 ng/ml) and or anti-TNF antibody (1 or 10 ug/ml) in a colony formation assay (MethoCult; StemCell) and then examined their colony growth, immunophenotypic characteristics and apoptosis over a period of 12 days. Where indicated serial re-plating was performed. Expression of apoptotic regulators was assessed by qRT-PCR. Results: In our initial triplicate experiments, starting with equal amount of Tet2 +/+ and -/- Lin- cells (104 cells/MethoCult well), cultured in appropriate growth factors and graded concentrations of TNFα , we observed that Tet2 -/- Lin- cells displayed superior colony forming ability over +/+ in serial re-plating assays under stress of increasing TNFα. To further confirm these initial results, we cultured the cells in-vitro Blocking Assays. We observed that in-vitro blockade of TNF-α with anti-TNF antibody, significantly limits the colony forming superiority of re-plated Tet2 -/- progenitors over +/+ in a dose dependent manner. (P<0.05). Immunophenotypic analysis of the colonies after replating showed significantly increased proportion of Mac1+Gr1+ and Sca1+Kit1+ populations in Tet2-/-, as compared to +/+, in the presence of TNF-α (P<0.05). These findings suggest that deregulation of innate and inflammatory cytokine signaling may provide a permissive environment for clonal dominance of Tet2 mutant HSPCs with an associated myeloid bias. To address the mechanisms behind these observations, we examined the susceptibility of both wild type and -/- cells to apoptosis using an annexin V- propidium iodide flow assay and then measured their apoptotic index. Our results showed that Tet2 -/- HSPCs have a lower apoptotic index compared to +/+ under stress of increasing TNF-α, suggesting that Tet2-/- BM cells may be resistant to TNFα induced apoptosis. Next, we searched for components of TNFα signaling pathway that might have been altered in resistant Tet2 -/- clones. Specifically, we measured the fold change in mRNA expression levels (using SYBR green qRT-PCR method) of some pro-apoptotic (TNFR1 and II, Fas-R, caspase 3 and 8) and anti-apoptotic genes (BCL-2, BCL-XL, IAP1, IAP2,) in both +/+ and -/- progenitor cells that have been cultured +/-TNF-α. Our results showed that mRNA levels of TNFR1, Fas receptors and caspase 8 were significantly lower in Tet2 -/- cells, compared to +/+ under inflammatory stress of TNF-α (P<0.05). In contrast, the mRNA levels of BCL2, IAP1 and IAP2 were significantly elevated in Tet2 -/- compared to +/+ (P<0.05). We also measured TNFα mRNA expression levels. Our preliminary data showed increased expression of TNF α mRNA in Tet2 -/-progenitors, compared to +/+ in absence of TNF-α (P<0.05). We also examined colony growths of TET2 mutant and non-mutant human MDS under varying TNF-α concentrations, using frozen BM mononuclear cells (BMMNCs) from MDS patients and normal healthy controls. Colony formation at 0.0 ng/ml TNFα was normalised to 100%. Our preliminary data showed that BFU-E colonies in TET2 mutant MDS show relatively increased survival at high TNFα concentrations, compared to wild type and controls, while CFU-GM colony formation in Tet2 mutant MDS was enhanced by low TNFα concentrations (P< 0.05). We are currently examining gene signatures in these groups of patients. Conclusion: Our data introduce a model where Tet2 mutation promotes clonal dominance by conferring TNFα resistance to TNFα sensitive progenitors, while also generating a TNFα rich environment. Mutations that promote resistance to environmental stem cell stressors are a known mechanism of clonal selection in aplastic anaemia and JAK2-mutant MPN and our findings suggest that this mechanism may be critical to clonal selection and dominance in MDS. Disclosures No relevant conflicts of interest to declare.

Differential humoral and cellular immunity induced by vaccination using plasmid DNA and protein recombinant expressing the NS3 protein of dengue virus type 3.

BackgroundThe dengue non-structural 3 (NS3) is a multifunctional protein, containing a serine-protease domain, located at the N-terminal portion, and helicase, NTPase and RTPase domains present in the C-terminal region. This protein is considered the main target for CD4+ and CD8+ T cell responses during dengue infection, which may be involved in protection. However, few studies have been undertaken evaluating the use of this protein as a protective antigen against dengue, as well as other flavivirus. In the present work we evaluated the potential of the NS3 (protease domain) as a protective antigen by comparing the administration of a recombinant protein versus a DNA vaccine in the mouse model.ResultsBALB/c mice were immunized with the recombinant protein NS3-DEN3 via intraperitoneal and with plasmid pcDNA3/NS3-DEN3 intramuscularly and the immune response was evaluated. The activity of T lymphocytes was analyzed by the MTT assay, and cells of mice immunized with the recombinant protein showed no activity when stimulated with the homologous protein. However, cells from mice immunized with DNA, responded to stimulation with the recombinant protein. When the expression (RT-PCR) and cytokine production (ELISA) was evaluated in the splenocytes, different behavior depending on the type of immunization was observed, splenocytes of mice immunized with the recombinant protein expressed cytokines such as IL-4, IL-10 and produced high concentrations of IL-1, IL-6 and TNFα. Splenocytes from mice immunized with DNA expressed IL-2 and IFNγ and did not produce IL-6. In addition, immunization with the recombinant protein induced the production of antibodies that are detected up to a dilution 1:3200 by ELISA and Western blot assays, however, the serum of mice immunized with DNA presented no detectable antibody titers.ConclusionThe results obtained in this study show that administration of pcDNA3/NS3-DEN3 induces a favorable response in the activation of T lymphocytes with low production of specific antibodies against NS3-DEN3.

Loss of XIAP facilitates switch to TNFα-induced necroptosis in mouse neutrophils.

Neutrophils are essential players in the first-line defense against invading bacteria and fungi. Besides its antiapoptotic role, the inhibitor of apoptosis protein (IAP) family member X-linked IAP (XIAP) has been shown to regulate innate immune signaling. Whereas the role of XIAP in innate signaling pathways is derived mostly from work in macrophages and dendritic cells, it is not known if and how XIAP contributes to these pathways in neutrophils. Here we show that in response to bacterial lipopolysaccharides (LPS), mouse neutrophils secreted considerable amounts of tumor necrosis factor-α (TNFα) and interleukin-1β (IL-1β) and, in accordance with earlier reports, XIAP prevented LPS-induced hypersecretion of IL-1β also in neutrophils. Interestingly, and in contrast to macrophages or dendritic cells, Xiap-deficient neutrophils were insensitive to LPS-induced cell death. However, combined loss of function of XIAP and cIAP1/-2 resulted in rapid neutrophil cell death in response to LPS. This cell death occurred by classical apoptosis initiated by a TNFα- and RIPK1-dependent, but RIPK3- and MLKL-independent, pathway. Inhibition of caspases under the same experimental conditions caused a shift to RIPK3-dependent cell death. Accordingly, we demonstrate that treatment of neutrophils with high concentrations of TNFα induced apoptotic cell death, which was fully blockable by pancaspase inhibition in wild-type neutrophils. However, in the absence of XIAP, caspase inhibition resulted in a shift from apoptosis to RIPK3- and MLKL-dependent necroptosis. Loss of XIAP further sensitized granulocyte–macrophage colony-stimulating factor (GM-CSF)-primed neutrophils to TNFα-induced killing. These data suggest that XIAP antagonizes the switch from TNFα-induced apoptosis to necroptosis in mouse neutrophils. Moreover, our data may implicate an important role of neutrophils in the development of hyperinflammation and disease progression of patients diagnosed with X-linked lymphoproliferative syndrome type 2, which are deficient in XIAP.

Lymphocytic, cytokine and transcriptomic profiles in peripheral blood of dogs with atopic dermatitis.

BackgroundCanine atopic dermatitis (cAD) is a common chronic and pruritic skin disease in dogs. The development of cAD involves complex interactions between environmental antigens, genetic predisposition and a number of disparate cell types. The aim of the present study was to perform comprehensive analyses of peripheral blood of AD dogs in relation to healthy subjects in order to determine the changes which would be characteristic for cAD.ResultsThe number of cells in specific subpopulations of lymphocytes was analyzed by flow cytometry, concentration of chosen pro- and anti-inflammatory cytokines (IL-4, IL-10, IL-13, TNF-α, TGF-β1) was determined by ELISA; and microarray analysis was performed on RNA samples isolated from peripheral blood nuclear cells of AD and healthy dogs. The number of Th cells (CD3+CD4+) in AD and healthy dogs was similar, whereas the percentage of Tc (CD3+CD8+) and Treg (CD4+CD25+ Foxp3+) cells increased significantly in AD dogs. Increased concentrations of IL-13 and TNF-α, and decreased levels of IL-10 and TGF-β1 was observed in AD dogs. The level of IL-4 was similar in both groups of animals. Results of the microarray experiment revealed differentially expressed genes involved in transcriptional regulation (e.g., transcription factors: SMAD2, RORA) or signal transduction pathways (e.g., VEGF, SHB21, PROC) taking part in T lymphocytes lineages differentiation and cytokines synthesis.ConclusionsResults obtained indicate that CD8+ T cells, beside CD4+ T lymphocytes, contribute to the development of the allergic response. Increased IL-13 concentration in AD dogs suggests that this cytokine may play more important role than IL-4 in mediating changes induced by allergic inflammation. Furthermore, observed increase in Treg cells in parallel with high concentrations of TNF-α and low levels of IL-10 and TGF-β1 in the peripheral blood of AD dogs point at the functional insufficiency of Treg cells in patients with AD.Electronic supplementary materialThe online version of this article (doi:10.1186/s12917-016-0805-6) contains supplementary material, which is available to authorized users.

Tumor Necrosis Factor-α Attenuates the Osteogenic Differentiation Capacity of Periodontal Ligament Stem Cells by Activating PERK Signaling.

Human periodontal ligament stem cells (PDLSCs) display efficient osteogenic differentiation capacity but fail to rescue bone breakdown associated with periodontitis. Endoplasmic reticulum (ER) stress and the unfolded protein response have recently been linked to inflammation and osteogenic differentiation. Therefore, the role of the double-stranded RNA-activated protein kinase (PKR)-like ER kinase (PERK) pathway in the impaired osteogenic differentiation ability of PDLSCs treated with tumor necrosis factor (TNF)-α was investigated. PDLSCs were isolated and stimulated with osteogenic media containing 1, 10, or 20 ng/mL TNF-α. Assessment included: 1) expression of runt-related transcription factor 2 and osteocalcin; 2) mRNA expression and activity of alkaline phosphatase; and 3) formation of mineralization nodules. Furthermore, expression of PERK pathway-related factors: 1) glucose-regulated protein (GRP) 78; 2) PERK; 3) activating transcription factor (ATF) 4; and 4) CCAAT-enhancer-binding proteins (C/EBP) homologous protein were also measured. Osteogenic differentiation and inhibition of the PERK pathway were also examined in cells pretreated with an inhibitor of ER stress, 4-phenylbutyric acid (PBA), followed by TNF-α stimulation. Finally, PERK small interfering RNA was used to examine osteogenic differentiation attenuated by TNF-α. Higher concentrations of TNF-α (10 and 20 ng/mL) impaired osteogenic differentiation of PDLSCs but activated the PERK pathway. Pretreatment of PDLSCs with lower concentrations of 4-PBA prevented the TNF-α-induced upregulation of GRP78, PERK, and ATF4 and recovered differentiation ability. Finally, PERK knockdown also restored osteogenic differentiation. TNF-α attenuates osteogenic differentiation ability of PDLSCs through activation of the PERK pathway.

A biphasic effect of TNF-α in regulation of the Keap1/Nrf2 pathway in cardiomyocytes

Antagonizing TNF-α signaling attenuates chronic inflammatory disease, but is associated with adverse effects on the cardiovascular system. Therefore the impact of TNF-α on basal control of redox signaling events needs to be understand in more depth. This is particularly important for the Keap1/Nrf2 pathway in the heart and in the present study we hypothesized that inhibition of a low level of TNF-α signaling attenuates the TNF-α dependent activation of this cytoprotective pathway. HL-1 cardiomyocytes and TNF receptor1/2 (TNFR1/2) double knockout mice (DKO) were used as experimental models. TNF-α (2–5ng/ml, for 2h) evoked significant nuclear translocation of Nrf2 with increased DNA/promoter binding and transactivation of Nrf2 targets. Additionally, this was associated with a 1.5 fold increase in intracellular glutathione (GSH). Higher concentrations of TNF-α (>10–50ng/ml) were markedly suppressive of the Keap1/Nrf2 response and associated with cardiomyocyte death marked by an increase in cleavage of caspase-3 and PARP. In vivo experiments with TNFR1/2-DKO demonstrates that the expression of Nrf2-regulated proteins (NQO1, HO-1, G6PD) were significantly downregulated in hearts of the DKO when compared to WT mice indicating a weakened antioxidant system under basal conditions. Overall, these results indicate that TNF-α exposure has a bimodal effect on the Keap1/Nrf2 system and while an intense inflammatory activation suppresses expression of antioxidant proteins a low level appears to be protective.

Serum and bile cytokines dynamics in patients with non-tumoral obstructive jaundice and suppurative cholangitis

To study serum and bile cytokines dynamics in patients with non-tumoral obstructive jaundice and suppurative cholangitis. Comparative study of serum and bile cytokines in 49 operated patients with biliary pathology was performed. Patients were divided into 3 groups. The first group included 24 patients with acute calculous cholecystitis (ACCh), the second group consisted of 12 patients with chronic calculous cholecystitis (CCCh) and the third group included 13 patients with choledocholithiasis complicated by obstructive jaundice and suppurative cholangitis. All patients had increased level of TNF-α, IL-4 и IL-6. There was augmentation of biliary cytokines in patients with obstructive jaundice compared with those with acute and chronic calculous cholecystitis. In patients with obstructive jaundice and suppurative cholangitis the highest levels of cytokines were observed. Patients with obstructive jaundice (OJ), suppurative cholangitis (SCh) and chronic calculous cholecystitis had higher levels of cytokines in both blood and bile than in those with OJ, SCh and acute calculous cholecystitis that is explained by development of purulent inflammation on background of previous cytokines imbalance and chronic inflammation. In general, patients with acute calculous cholecystitis had higher levels of TNF-α and IL-6 in blood serum and IL-4 in bile. Patients with chronic calculous cholecystitis had higher concentration of TNF-α and IL-4 in bile and IL-6 - in blood serum. Both forms of cholecystitis and their complications are associated with increased cytokines in serum and bile. The level of augmentation depends on the type of inflammation and complications. Comparative study of cytokines can be the most informative criterion to monitor the postoperative period.

Helicobacter pylori infection and gastric carcinoma: Not all the strains and patients are alike.

Gastric carcinoma (GC) develops in only 1%-3% of Helicobacter pylori (H. pylori) infected people. The role in GC formation of the bacterial genotypes, gene polymorphisms and host's factors may therefore be important. The risk of GC is enhanced when individuals are infected by strains expressing the oncoprotein CagA, in particular if CagA has a high number of repeats containing the EPIYA sequence in its C'-terminal variable region or particular amino acid sequences flank the EPIYA motifs. H. pylori infection triggers an inflammatory response characterised by an increased secretion of some chemokines by immunocytes and colonised gastric epithelial cells; these molecules are especially constituted by proteins composing the interleukin-1beta (IL-1β) group and tumour necrosis factor-alpha (TNF-α). Polymorphisms in the promoter regions of genes encoding these molecules, could account for high concentrations of IL-1β and TNF-α in the gastric mucosa, which may cause hypochlorhydria and eventually GC. Inconsistent results have been attained with other haplotypes of inflammatory and anti-inflammatory cytokines. Genomic mechanisms of GC development are mainly based on chromosomal or microsatellite instability (MSI) and deregulation of signalling transduction pathways. H. pylori infection may induce DNA instability and breaks of double-strand DNA in gastric mucocytes. Different H. pylori strains seem to differently increase the risk of cancer development run by the host. Certain H. pylori genotypes (such as the cagA positive) induce high degrees of chronic inflammation and determine an increase of mutagenesis rate, oxidative-stress, mismatch repair mechanisms, down-regulation of base excision and genetic instability, as well as generation of reactive oxygen species that modulate apoptosis; these phenomena may end to trigger or concur to GC development.

Dynamic Change and Impact of Circulating Myeloid-Derived Suppressor Cells in Patients Following Allogeneic Hematopoietic Stem Cell Transplantation

Myeloid-derived suppressor cells (MDSCs) are a heterogeneous cell population involved in tumor-associated immunosuppression and played immune regulatory roles in malignancies, infectious disease, autoimmune disease, trauma and inflammatory disease. Based on the significant suppressive functions, MDSCs raises great interests in the field of allogenetic hematopoietic stem cell transplantation (allo-HSCT) and graft-versus-host disease (GVHD). In recent years, studies described the immunosuppressive functions of CD14+HLA-DR-/low MDSCs, one of the few well-characterized MDSC subsets in human. However, the studies in patients with allo-HSCT are still scanty. This study identified the phenotype of CD14+HLA-DR-/low MDSCs and systematically monitored the dynamic changes of MDSCs accumulation in patients during the first 100 days after allo-HSCT in order to evaluate possible effects of MDSCs on aGVHD development and clinical outcomes. We also detected the frequencies of other cell types and concentrations of relative cytokines during MDSCs accumulation. Results showed that accumulation of MDSCs in the graft and in peripheral blood when engraftment might contribute to patients' overall immune suppression and result in the successful control of severe aGVHD and long-term survival without influence on risk of recurrence after allo-HSCT. However, the level of MDSCs in the graft had more favorable predictive abilities compared with that in PBMCs when engraftment. Furthermore, MDSCs frequencies were significantly increased in patients developing aGVHD after allo-HSCT.In the patients with mild aGVHD (0-2), MDSCs accumulated at the time of engraftment after allo-HSCT and decreased to basal levels at about 4 weeks. MDSCs frequencies would keep in stable levels with slight fluctuations in the following weeks. But, in patients with severe aGVHD (3-4), MDSCs elevated slightly when engraftment. When aGVHD occurred, MDSCs frequencies would significantly increase. And a synchronized reduction of the levels of MDSCs was observed after effective management of aGVHD with immunosuppressive therapy. Besides, the levels of IL-10, IL-6, TNF-α, Arg-1, iNOS and HO-1 increased greatly both in patients with aGVHD and in high MDSCs group. It is speculated that MDSC accumulation at the onset of aGVHD might be caused by secondary inflammatory response, especially related to high concentrations of IL-6 and TNF-α. But the accumulation would not be able to counterbalance the aggravation of aGVHD and would not have influence on clinical outcomes and risk of relapse in future. Overall, we suggested that MDSCs might be considered as a potential new approaches to regulate transplant rejection and achieve the recipient host long-term acceptance and survival. [Display omitted] [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Age‐associated changes in long‐chain fatty acid profile during healthy aging promote pro‐inflammatory monocyte polarization via PPAR γ

Differences in lipid metabolism associate with age-related disease development and lifespan. Inflammation is a common link between metabolic dysregulation and aging. Saturated fatty acids (FAs) initiate pro-inflammatory signalling from many cells including monocytes; however, no existing studies have quantified age-associated changes in individual FAs in relation to inflammatory phenotype. Therefore, we have determined the plasma concentrations of distinct FAs by gas chromatography in 26 healthy younger individuals (age<30years) and 21 healthy FA individuals (age>50years). Linear mixed models were used to explore the association between circulating FAs, age and cytokines. We showed that plasma saturated, poly- and mono-unsaturated FAs increase with age. Circulating TNF-α and IL-6 concentrations increased with age, whereas IL-10 and TGF-β1 concentrations decreased. Oxidation of MitoSOX Red was higher in leucocytes from FA adults, and plasma oxidized glutathione concentrations were higher. There was significant colinearity between plasma saturated FAs, indicative of their metabolic relationships. Higher levels of the saturated FAs C18:0 and C24:0 were associated with lower TGF-β1 concentrations, and higher C16:0 were associated with higher TNF-α concentrations. We further examined effects of the aging FA profile on monocyte polarization and metabolism in THP1 monocytes. Monocytes preincubated with C16:0 increased secretion of pro-inflammatory cytokines in response to phorbol myristate acetate-induced differentiation through ceramide-dependent inhibition of PPARγ activity. Conversely, C18:1 primed a pro-resolving macrophage which was PPARγ dependent and ceramide dependent and which required oxidative phosphorylation. These data suggest that a midlife adult FA profile impairs the switch from proinflammatory to lower energy, requiring anti-inflammatory macrophages through metabolic reprogramming.

The Effect of Tumor Necrosis Factor-α at Different Concentrations on Osteogenetic Differentiation of Bone Marrow Mesenchymal Stem Cells.

Tumor necrosis factor-alpha (TNF-α) has been demonstrated to have a close relationship with inflammation in the body. Although most researchers confirmed that TNF-α can have an effect on the expression of osteoblast specific genes, they had not elucidated the regulation of inflammatory factors in osteogenic gene expression during the process of bone marrow mesenchymal stem cells (BMMSCs) differentiating to osteoblast. The aim of this study was to investigate the effect of TNF-α at different concentrations on osteogenetic differentiation of BMMSCs. In this study, BMMSCs proliferation was analyzed by using cell counting kit-8 assay, cell osteogenic differentiation was evaluated by means of alkaline phosphatase activity assay and Von Koaas staining and the messenger RNA (mRNA) expression of bone morphogenetic proteins-2 (BMP-2) and drosophila mothers against decapentaplegic protein 1 (Smad1) was measured through real-time polymerase chain reaction. The results indicated that a low concentration of TNF-α at short-term promotes the osteogenetic differentiation of BMMSCs and increases the mRNA expression of BMP-2 and Smad1, but inhibits the osteogenetic differentiation of BMMSCs and the expression of BMP-2 and Smad1 at long term. In addition, regardless of a short or long time, a high concentration of TNF-α inhibits the osteogenetic differentiation of BMMSCs and the expression of Smad1, but results in a high expression of BMP-2.

Niveles séricos de marcadores inflamatorios en pacientes hipertensos de la E.S.E. Imsalud, Cúcuta, Colombia

Introducción: la hipertensión arterial es una condición clínica multifactorial, caracterizada por la elevación de la presión arterial sistólica o diastólica, establecida como un factor de riesgo importante en la aparición de enfermedades cardiovasculares. Objetivos: determinar los niveles séricos de IFN-γ, TNF-α y proteína C reactiva, relacionándolos con la hipertensión arterial en pacientes que asisten a la E.S.E. Imsalud-Unidad Materno Infantil la Libertad (Cúcuta, Colombia). Materiales y métodos: se realizó un estudio descriptivo, correlacional en individuos hipertensos y controles sanos seleccionados mediante encuesta estructurada. Los niveles séricos del IFN-γ y el TNF-α fueron analizados por ELISA y los de la proteína C reactiva ultrasensible por inmunoturbimetría. Resultados: el 52,8% y el 29,3% de los hipertensos presentaron concentraciones moderadamente altas de IFN-γ y TNF-α respectivamente. De acuerdo a los niveles séricos de proteína C reactiva ultrasensible, el 81,2% de los hipertensos se ubicaron en riesgo cardiovascular bajo, el 6,2% en medio y el 12,5% en alto. Conclusiones: los niveles elevados de IFN-γ en los pacientes hipertensos respecto a los controles sugieren que esta citoquina puede ser considerada un marcador útil como herramienta de evaluación del pronóstico de desarrollo de complicaciones cardiovasculares. Aunque el TNF-α no se encontró aumentado en los pacientes hipertensos, podría ser empleado en aquellos con difícil control de la enfermedad o en quienes no han iniciado el tratamiento antihipertensivo. La proteína C reactiva ultrasensible se reitera como un marcador útil en la estratificación de los niveles de riesgo en los hipertensos, abarcando desde bajo hasta alto de afectación cardiovascular.

Concentrações de IL-6, TNF-α e MCP-1 em crianças com excesso de massa corporal

O objetivo desta revisão sistemática foi verificar a relação das concentrações de IL-6, TNF-α e MCP-1 em crianças com excesso de massa corporal. As bases de dados investigadas PUBMED, SciELO, LILACS e Periódico Capes foram consultadas retrospectivamente para os últimos seis anos (2009 a 2014) utilizando combinações de palavras chaves como inflamação, IL-6, TNF-α, MCP-1 combinadas com crianças e escolares. Foram analisados 21 artigos. Foi encontrado associação entre sobrepeso/obesidade com IL-6, TNF-α e MCP-1 em 73,3% (11/15), 80% (12/15) e 60% (3/5) dos estudos que analisaram tais marcadores, respectivamente. Crianças com excesso de massa corporal possuem concentrações elevadas de IL-6, TNF-α e MCP-1 resultando em inflamação sistêmica crônica e aumentando o risco de desenvolvimento de outras doenças cardiovasculares.

Pro-inflammatory interleukins in middle ear effusions from atopic and non-atopic children with chronic otitis media with effusion.

Chronic otitis media with effusion (OME) is associated with irreversible changes in the middle ear, sometimes leading to hearing loss and abnormal language development in children. While the pathogenesis of OME is not fully understood, inflammatory and allergic factors are thought to be involved. The study aimed to investigate the role of cytokines in the local development of chronic OME, and assess differences in the cytokine profiles between atopic and non-atopic children. 84 atopic and non-atopic children with chronic OME (mean age of 6years 7months) were studied. Age-matched children with hypertrophy of the adenoids and Eustachian tube dysfunction served as the control group. The number of past acute otitis media (AOM) episodes, their age, and the type of effusion were recorded for all children. Pro-inflammatory cytokine concentrations (TNF-α, IL-1β, IL-6 and IL-8) were determined and the presence of pathogenic bacteria in the patients' effusions was examined. High concentrations of TNF-α, IL-1β, IL-6 and IL-8 were found in the effusions in all children with chronic OME, with the highest levels observed in the non-atopic group. The atopic group showed persistently high IL-1β levels, while in the non-atopic children, IL-1β and TNF-α levels positively correlated with the patient's age and the number of past AOM episodes. Pathogenic bacteria were more frequently isolated from effusions in non-atopic children. In both atopic and non-atopic children, pro-inflammatory cytokines are found at high concentrations. This argues in favor of instituting anti-inflammatory management for treating OME, regardless of atopy.

Obstructive sleep apnea exacerbates airway inflammation in patients with chronic obstructive pulmonary disease

BackgroundChronic obstructive pulmonary disease (COPD) concomitant with obstructive sleep apnea (OSA) is known as overlap syndrome. It has an increased rate of hospitalization due to COPD exacerbation which is believed to indicate a worsening of the underlying chronic airway inflammation. Therefore, the aim of this prospective study was to explore whether OSA exacerbates airway inflammation in subjects with COPD by examining the bronchoalveolar lavage (BAL) fluid. MethodsThis prospective study included 47 patients with overlap syndrome and 28 patients with moderate-to-severe stage stable COPD. Twenty-five patients with overlap syndrome adhered to the continuous positive airway pressure (CPAP) treatment; the remaining patients either refused CPAP treatment or discontinued it within two weeks owing to adverse effects or other reasons. BAL fluid was collected from all subjects for the evaluation of cell numbers and tumor necrosis factor alpha (TNFα) and interleukin-8 (IL-8) levels. ResultsThe BAL fluid of patients with overlap syndrome showed a significantly increased proportion of neutrophils and higher TNFα concentration and IL-8 levels than that of COPD patients; however, the serum CRP levels were not significantly different. An association was found between the percentage of neutrophils and the TNFα concentration and IL-8 levels. Moreover, the TNFα concentration was significantly correlated with the percentage of nighttime spent with oxygen saturation less than 90%. After CPAP treatment, airway inflammation was found to decrease significantly. ConclusionsOSA exacerbates airway inflammation in COPD patients. CPAP treatment can improve nocturnal hypoxemia and decrease airway inflammation.

High Concentrations of TNF-α Induce Cell Death during Interactions between Human Umbilical Cord Mesenchymal Stem Cells and Peripheral Blood Mononuclear Cells

Human umbilical cord mesenchymal stromal cells (hUC-MSCs) are currently being used as novel therapeutic agents in numerous clinical trials. Previous works have shown that hUC-MSCs possess profound immunomodulatory capacities through IL-1 stimulation produced by peripheral blood mononuclear cells (PBMCs), their main cellular partner in most pathophysiological and therapeutic situations. The present study was designed to explore the role of TNF-α in these interactions. In these experiments, we demonstrated that TNF-α originated from PBMCs under the influence of IL-1. We also showed that TNF-α acted differently depending upon the concentrations reached. At low concentrations it clearly contributed to IL-6 and monocyte chemotactic protein 1 (MCP-1) production. At high concentrations, used alone or in association with the TNF-related apoptosis-inducing ligand, TNF-α also stimulated hUC-MSC IL-6 but, more intensely, MCP-1 production. This stimulation was associated but independent of apoptosis induction in a process involving Inhibitor of Apoptosis Proteins. Interferon gamma (IFN-γ), tested to stimulate PBMC and tissue activation, amplified IL-6 and MCP-1 production and cell death by, apparently, a different process involving necrosis. Our findings bring new insights into the complex interactions between hUC-MSCs and PBMCs, involving cytokines, chemokines and cell death, and are of fundamental importance for tissue homeostasis.

Expression of interleukin-34 and colony stimulating factor-1 in the stimulated periodontal ligament cells with tumor necrosis factor-α.

Tumor necrosis factor-α (TNF-α) directly and indirectly plays a crucial role in osteoclastogenesis. However, the indirect effects of TNF-α on colony-stimulating factor-1 receptor (CSF-1R)-mediated osteoclastogenesis achieved via periodontal ligament (PDL) cells are not fully understood. We herein examined the potency of osteoclast differentiation and maturation induced by fivefold supernatants in the stimulated human PDL cells with a physiologically high concentration (10 ng/mL) of recombinant TNF-α to human peripheral blood monocytes/macrophages in the simultaneous presence of the receptor activator of nuclear factor kappa-B ligand. The number of tartrate-resistant acid phosphatase-positive cells with multiple nuclei, but not those with a single nucleus, was decreased by approximately 50% by neutralization with rabbit IgG against either interleukin-34 (IL-34) or CSF-1. Small and large amounts of IL34 and CSF1 transcripts were measured in the stimulated PDL cells using real-time polymerase chain reaction. The corresponding amounts of proteins to IL34 and CSF1 transcripts were observed in the stimulated PDL cells on immunohistochemical staining or Western blotting. Moreover, 0.13 ng/mL of IL-34 and 5.0 ng/mL of CSF-1 were measured in the supernatants of the stimulated PDL cells using an enzyme-linked immunosorbent assay. IL-34 derived from the stimulated PDL cells with TNF-α appeared to synergistically function with CSF-1 in the CSF-1R-mediated maturation of osteoclastogenesis.