TNF-α Expression In Patients Research Articles

Seipin is involved in oxygen-glucose deprivation/reoxygenation induced neuroinflammation by regulating the TLR3/TRAF3/NF-κB pathway

Seipin plays a crucial role in lipid metabolism and is involved in neurological disorders. However, the function and mechanism of action of seipin in acute ischemic stroke have not yet been elucidated. Here, we aimed to investigate the effect of seipin on neuroinflammation induced by oxygen-glucose deprivation/reoxygenation (OGD/R) and further explore the molecular mechanism by functional experiments. Our results revealed a significant decrease in seipin mRNA levels, accompanied by enhanced expression of TNF-α in patients with AIS, and a significant negative correlation between seipin and TNF-α was observed. Additionally, there was a negative correlation between seipin levels and the National Institutes of Health Stroke Scale (NIHSS) score. Furthermore, seipin levels were also decreased in middle cerebral artery occlusion/reperfusion (MCAO/R) mice and OGD/R-treated BV2 cells. RNA sequencing analysis showed that seipin knockdown altered the Toll-like receptor 3 (TLR3) signaling pathway. It was further confirmed in vitro that seipin knockdown caused significantly increased secretion of inflammatory factors including TNF-α, interleukin (IL)-1β, and interferon (IFN)-β. Meanwhile, seipin knockdown activated the Tlr3 signal pathway while this effect could be reversed by Tlr3 inhibitor in OGD/R treated BV2 cells. Furthermore, neuroinflammation induced by OGD/R was significantly reduced by seipin overexpression. Overall, our study demonstrate that seipin deficiency aggravates neuroinflammation by activating the TLR3/TRAF3/NF-κB signaling pathway after OGD/R stimuli, and suggest that seipin may be a potential therapeutic target for AIS.

Exploring the Relationship between TNF-α Gene Expression in Non Diabetic Nephropathy Type 2 Diabetes Patients

Non-diabetic nephropathy (NDN) is a common complication of type 2 diabetes, leading to kidney damage and impaired kidney function. TNF-α (tumor necrosis factor-alpha) is a pro-inflammatory cytokine that has been implicated in the development of NDN. Objective: To evaluate the gene expression of TNF-α in patients with type 2 diabetes mellitus (T2DM) without nephropathy to gain insight into the potential role of TNF-α in the pathogenesis of diabetic nephropathy (DN). Methods: Total of 80 subjects were tested, split into two groups, including healthy patients, T2DM patients without nephropathy, and T2DM patients with nephropathy. RNA was extracted from blood samples, and RT-PCR was used to observe the impact of T2DM without nephropathy on the expression of the TNF-α gene using gene-specific primers and SYBR Green mix. Results: The results showed almost 4.4-fold induced expression of TNF-α in T2DM patients without nephropathy compared to the normal group. Conclusion: The findings may have implications for the development of new therapies and biomarkers for DN, and for a good interpretation of the complex pathophysiology of T2DM. The exact role of TNF-α in the pathogenesis of DN in humans is not fully understood, and further investigation is needed.

Abstract TP343: Differential TNFα Expression in Patients of Acute Ischemic Stroke

Introduction: Inflammation plays a critical role in the pathophysiology of ischemia and atherosclerosis. TNFα plays a multifaceted role in stroke progression, having both protective and deleterious effects. Animal studies have shown that TNFα levels increase in cerebral tissue within the first 24 hours after experimental stroke. Hypothesis: Expression of TNFα is related to outcomes after acute ischemic stroke. Differences in expression will be seen with respect to patient gender and other stroke characteristics. Methods: Patients admitted to Hartford Hospital between January 2011 and March 2014 were considered for this study. Blood samples were collected within 24 hours of stroke presentation. Patients with a history of active cancer, primary or secondary neoplastic brain lesions, traumatic brain injury or intracranial hemorrhage were excluded from the study. Serum was analyzed using a multiplex human cytokine panel/Bio plex (Bio-Rad laboratories Inc.). Statistical analysis was performed using SPSS. Results: A total of 163 ischemic stroke patients were included in the study. Outcome variables including NIH stroke scale, modified Rankin scale (mRS) at discharge, 3 months, 12 months and modified Barthel’s index (BI) were collected. Composite outcome scores were determined using the mRS and BI. A negative outcome was defined as death or BI ≤ 14 or mRS >2.There was a statistically significant difference (p=0.003) in the expression of TNFα between male - 21.74 pg/ml (15.11-30.21) and female 27.9 pg/ml (19.07-35.497) stroke patients. A significant difference (p=0.009) was observed in TNFα expression in patients with a good outcome - 22.96 pg/ml (15.69-30.85) as compared to those with a poor outcome (discharge to worse/hospice or death) - 30.78 pg/ml (22.03-35.38) post stroke. Conclusions: Male- female sex differences are associated with changes in the post ischemic inflammatory cascade. Higher TNFα levels predict poorer outcomes post stroke. Cytokines like TNFα may serve as a potential therapeutic targets in patients with acute ischemic stroke.

Lack of association among TNF-α gene expression, -308 polymorphism (G > A) and virulence markers of Helicobacter pylori.

BackgroundTumor necrosis factor plays a critical role in the pathogenesis of gastric diseases such as gastric cancer, and an abnormal inflammatory response has frequently been observed in dyspeptic patients. Helicobacter pylori infection can induce a gastric mucosal inflammatory response that may be influenced by -308 (G > A) polymorphisms and gene expression of the TNF-α gene.MethodsOne hundred and thirty-four gastric biopsy samples were collected from patients of both genders (61♂ and 73♀, mean age 40.3 ± 24.2 years) with gastric symptoms. The -308 (G > A) polymorphism of TNF-α was characterized using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). The expression level was measured using real-time PCR, and relative quantification (RQ) was calculated using the comparative CT method (2-ΔΔCT).ResultsThe analysis revealed an increase in TNF-α gene expression in patients with gastritis; on the other hand, no statistical differences were observed in patients with gastric cancer. In addition, no association was found among -308 polymorphism genotypes, virulence markers, or TNF-α gene expression.ConclusionsHelicobacter pylori induces a large increase in TNF-α expression in patients with gastritis, regardless of tissue inflammation, but after the tissue becomes neoplastic, the presence of bacteria did not influence expression. These results suggest that the TNF-α pathway may play an important role in the progression from gastritis to gastric cancer

Influence of the naturally occurring human CASP1 variant L265S on subcellular distribution and pyroptosis

Patients with unexplained recurrent febrile episodes and CASP1 variants suffer from systemic sterile inflammation despite altered enzymatic activity of procaspase-1 and reduced IL-1β release. Most recent findings from our group indicate that the proinflammatory effects of CASP1 variants with reduced or abrogated enzymatic activity could be due to receptor interacting protein kinase 2 (RIP2) mediated increase of NF-kB activation. These findings are additionally supported by a trend to elevated IL-6 and TNF-α expression in patients with CASP1 variants.

JAK2V617F mutation and TNF-α expression in myeloproliferative neoplasms and their correlation

This study was aimed to explore the JAK2V617F mutation and TNF-α expression in patients with myeloproliferative neoplasm (MPN), and the relation between them so as to provide theoretical basis for clinical practice and target therapy. Sixty-two confirmed BCR-ABL-negative MPN patients and 15 healthy adults were enrolled in this study. The peripheral blood mononuclear cells of the patients and healthy controls were divided into two parts, one part was used to extract DNA, the other one was used to extract mRNA and reverse-transcribe into cDNA. Real-time fluorescent quantitative PCR was used to detect JAK2V617F mutation proportion and the expression level of TNF-α. The results showed that the positive rate of JAK2V617F mutation in MPN patients was 64.52% (40/62) , including 54.28% in essential thrombocythemia (ET) patients (19/35), 94.74% in polycythemia vera (PV) patients (18/19) and 37.50% in myelofibrosis (MF) (3/8) patients. Mutation proportions of JAK2V617F in ET, PV and MF patients were 0.838 ± 0.419, 4.417 ± 0.658, 2.746 ± 2.009 respectively. The expression of TNF-α in ET, PV and MF patients were higher than that in healthy controls: 1.7, 7.0, 8.2-fold (P < 0.05) respectively. In addition, TNF-α expression was correlated with JAK2V617F allele burden (Pearson r = 0.610,R(2) = 0.372,P = 0.005). It is concluded that TNF-α plays an important role in the pathogenesis of MPN, the TNF-α expression increases and is different in ET,PV and MF patients,which correlates with JAK2V617F allele burden.

Expression of IL-2, IL-17 and TNF-alpha in patients with Crohn's disease treated with anti-TNF antibodies

The T cell cytokine IL-17 and the Th-17 pathway appear to have a role in the pathogenesis of inflammatory bowel diseases. IL-2 is a potent stimulator of lymphocyte proliferation and IL2/IL21 receptor polymorphisms have recently been associated with susceptibility to IBD. To evaluate the expression of IL-17, IL-2 and TNFα in Crohn's disease (CD) patients with and without anti-TNFs. Cytokine expression was evaluated by ELISA and intracellular staining of CD4(+) T-cells from the peripheral blood and lamina propria of CD patients and of non-IBD controls. The results were stratified by disease activity and anti-TNF treatment. IL2 expression was significantly elevated in CD patients not treated with anti-TNFs in comparison to healthy controls (19.6% vs. 33.3%, P=0.03) and CD patients treated with anti-TNFs (20.4% vs. 33.3%, P=0.02), and similar in infliximab-treated patients and controls. IL17 expression was similar in CD patients and controls, and was not affected by anti-TNF therapy. TNFα expression in patients with active CD was increased compared to controls (35.5% vs 25.7%, P<0.005), and was significantly decreased in anti-TNF treated patients in comparison to CD patients without anti-TNFs (39.6% vs 26.2%, P=0.01). Expression of IL2 was significantly decreased in anti-TNF-treated CD patients in comparison to non-treated CD patients and controls. This novel finding may indicate a further mechanism of anti-TNF therapy in CD. Expression of IL17 was not influenced by presence of CD or anti-TNF therapy, which may partly explain the failure of recent clinical trials investigating anti-IL17 therapy in CD.

Cytokines Elicited by HSP60 in Periodontitis with and without Coronary Heart Disease

The human 60 kDa and microbial 65 kDa heat shock proteins (HSP) have been implicated in the pathogenesis of chronic periodontitis (CP) and coronary heart disease (CHD). We have studied 100 subjects: Group (a) consisted of patients with gingivitis (n = 25), group (b) were patients with CP (n = 25), group (c) patients with CHD and gingivitis (n = 25) and group (d) patients with CHD and CP (n = 25). PBMCs separated from peripheral blood were stimulated with medium, PMA/ionomycin, human HSP60, microbial HSP65, or no stimulus for 18 hours before intracellular IL-2, IFN-γ, TNF-α, IL-4, IL-5, or IL-17 were detected by flow cytometry. The mean fluorescence intensity (MFLI) for intracellular TNF-α was significantly increased when PBMC were stimulated with human HSP60 amongst the four groups (p = 0.001, ANOVA); pairwise comparisons revealed significant differences in MFLI between the gingivitis group and the CP (p = 0.017); between gingivitis and ging/CHD (p = 0.001) as well; but no significant difference between the CP and CP/CHD (p = 0.442). There was no significant difference in intracellular expression of IL-17, or any of the other cytokines tested; and the MFLI for HSP-stimulated were comparable to unstimulated cultures. When heat-labile human HSP60 was heated, intracellular cellular TNF-α expression was abrogated. In contrast, heat-stable LPS elicited TNF-α expression from monocytes in bulk cultures in all groups. These results suggest that the cytokine expression was dependent on human HSP60 and not LPS. Serum CRP was significantly associated with MFLI of intracellular TNF-α in CP patients (rs = 0.665, p = 0.026) and CP/CHD (rs = 0.699, p = 0.011). We conclude that human HSP60 elicits increased monocytic expression of TNF-α in patients with CP, CP/CHD or ging/CHD compared to patients with gingivitis. Since the marker of inflammation, namely CRP correlates with CP with or without CHD and not with mild chronic gingivitis or ging/CHD, this suggests that human HSP60-induced production of TNF-α is associated with CP and not CHD. There was no significant difference in intracellular expression of IL-17.

Inflammation: friend or foe of muscle remodelling in COPD?

An enhanced systemic inflammatory response is a typical feature of chronic diseases, including chronic obstructive pulmonary disease (COPD), cancer, rheumatoid arthritis and chronic heart failure. The pattern and degree of systemic cytokine elevation is remarkably comparable among these conditions, despite different primary organ pathology. Interestingly, these diseases share skeletal muscle weakness and wasting as a common phenomenon in the disease course, and experimental studies using different acute and chronic disease models have clearly implicated cytokine-mediated effects, particularly of tumour necrosis factor (TNF)-α, on muscle contractility and muscle atrophy and regeneration. TNF-α is thought to act on skeletal muscle cells in an endocrine as well as autocrine/paracrine fashion, but as yet it is unclear whether this cytokine is causally related to (altered) skeletal muscle remodelling and whether its effects facilitate or interfere with muscle plasticity in COPD. One approach to unravelling the role of TNF-α in atrophy, regeneration and muscle fibre type shifting is to compare local expression of TNF-α during muscle remodelling, e.g. in conditions which are accompanied by alterations in skeletal muscle mass, structure or metabolic activity. Limited studies have investigated TNF-α expression in muscle biopsies of patients with COPD and unfortunately, no studies are yet available comparing expression levels between different muscles within the same subject. Some studies 1, 2 have demonstrated elevated muscular TNF-α expression in patients with COPD compared with healthy control subjects, but this finding was not confirmed …

Invited commentary

Bittar and colleagues discuss whether the inflammatory response (particularly after coronary bypass operations) can be predicted by genetic phenotyping [1Bittar M.N. Carey J.A. Barnard J.B. et al.Tumor necrosis factor alpha influences the inflammatory response after coronary surgery.Ann Thorac Surg. 2006; 81: 132-138Abstract Full Text Full Text PDF PubMed Scopus (33) Google Scholar]. The bypass circuit (ie, blood foreign surface interface) causes an intense pro-inflammatory reaction. This response seems to be macrophage and endothelial-cell induced. The effect of tumor necrosis factor α (TNFα) by itself seems to be negative inotropic and pro-inflammatory. Some cytoprotective effects have been proposed as well. TNFα acts pro-inflammatory by self amplification as well as by induction and expression of other pro-inflammatory cytokines. The myocardium constitutively expresses a low level of TNFα. Although macrophages are activated at the blood foreign surface interface, an increase of myocardial TNF after ischemia and reperfusion has also been demonstrated. It is unclear if TNFα gene expression and peptide synthesis occur during global ischemia. Hence, the source of TNFα is multifactorial. It is intriguing to merge molecular genetics with molecular mechanisms of the systemic inflammatory response. However, the systemic inflammatory response involves a multitude of humeral and cellular mechanisms, and it would be an oversimplification to assume that one molecule is responsible for this very complex phenomenon. Genetic polymorphism is defined as the presence of multiple alleles at a gene locus in appreciable frequencies in a population. The TNF gene is embedded within the class three major histocompatibility complex on the short arm of chromosome 6. Whether TNF gene polymorphism influences TNFα expression in patients undergoing coronary artery bypass grafting with or without cardiopulmonary bypass has been the object of a number of studies. The results have been conflicting. How can we explain these discrepancies? First when planning or reading one of these studies, the goal at the onset has to be kept in mind. To study gene polymorphism and its possible influence on outcome as a consequence of cardiopulmonary bypass is one topic. To study the influence of gene polymorphism with two treatment strategies (ie, on pump vs off pump) is a completely different question. A third issue may be if gene polymorphism has the same distribution in a diabetic population as in a nondiabetic population. Furthermore, attention has to be paid to which allele is being compared. The TNFα gene contains a large number of alleles. As a clinical tool, genetic testing for the purpose of detecting which patients are at increased risk for postoperative complications is intriguing, but at present this is impractical for most of us. It is not clear at this time if TNFα is the marker that has the greatest predictive value or if another cytokine or molecular marker or a combination has more predictive value. However, as a concept, it is very appealing and will allow us to improve patient outcomes in the future, be it by tailoring of therapy in order to avoid complications, or be it by earlier recognition of the latter.

Enhanced expression of TNFα in patients with primary biliary cirrhosis

Primary biliary cirrhosis (PBC) is characterized by granuloma formation, plurivisceral involvement and the presence of a complex immune dysfunction. Tumor necrosis factor α (TNFα) is a cytokine endowed with powerful systemic actions which is believed to participate in granulomatous reactions. In this work we analyzed the expression of TNFα mRNA in resting peripheral blood mononuclear cells (PBMC) using a comparative PCR-based procedure and we determined the production of TNFα by stimulated and unstimulated PBMC using a bioassay method. TNFa mRNA levels (as a ratio to those of β-actin) were found to be higher in PBC patients (0.58 ± 0.03) than in normal controls (0.31 ± 0.02, P < 0.001) or patients with other liver diseases (0.31 ± 0.02,P < 0.001). Moreover, PBMC from PBC patients produced under LPS stimulation higher levels of bioactive TNFα (6.85 ± 1.44 U/ml) than normal controls (2.7 ± 0.59 U/ml,P < 0.05) or control patients (2.31 ± 0.96 U/ml,P < 0.05). No TNFα bioactivity was found in supernatants from unstimulated cells. In conclusion, our results show that there is an enhanced expression of TNFα gene in patients with PBC.