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Background: Receptor Expressed in Lymphoid Tissues (RELT) is a TNFRSF member that has two paralogs, RELL1 and RELL2; the three proteins are collectively referred to as RELT family members (RELTfms). Methods: We sought to evaluate RELT expression in cancerous cells by using real-time PCR, western blotting, flow cytometry, and immunohistochemistry (IHC). The mechanism of RELT-induced cell death was assessed by western blotting, flow cytometry, luciferase assays, and morphology staining. RELT localization was detected through immunofluorescence and western blotting, and co-immunoprecipitation was used to test whether a mutated RELT interacts with the OXSR1 kinase. Results: RELT and RELL1 protein expression was significantly elevated in cell lines representing breast and lung cancer, whereas RELL2 protein expression was relatively consistent across different cell lines. The surface expression of RELT was highest in monocytes. IHC staining revealed increased RELT expression in malignant breast cancer biopsies compared to patient-matched benign tissue. RELTfm overexpression induced death in MDA-MB-231 (231) breast cancer cells, accompanied by increased phosphatidylserine externalization and Caspase-3/7 activation. The co-transfection of plasmids predicted to block the phosphorylation of RELT by the OXSR1 kinase did not abrogate RELT-induced apoptosis, indicating that the activation of p38 by RELT through the OXSR1 kinase is not required for RELT-induced cell death. Interestingly, nuclear localization of RELT was detected in 231 and HEK-293 cells. Conclusions: These results demonstrate that RELT induces death in breast cancer cells through an apoptotic pathway that does not require OXSR1 phosphorylation and that RELT possesses the ability to translocate to the nucleus, a novel finding that warrants further investigation. [ABSTRACT FROM AUTHOR]

Background: Fatty acid-binding protein 4 (FABP4) is an adipokine that plays significant roles in the development of insulin resistance and atherosclerosis. High levels of soluble tumor necrosis factor receptors (TNFRs) including TNFR1 and TNFR2 are associated with renal dysfunction and increased mortality in patients with diabetes mellitus (DM). However, the association between circulating levels of FABP4 and TNFRs remains unclear. Methods: We investigated the associations of FABP4 with TNFRs and metabolic markers in Japanese patients with type 1 DM (T1DM, n = 76, men/women: 31/45) and type 2 DM (T2DM, n = 575, men/women: 312/263). Results: FABP4 concentration was positively correlated with levels of TNFR1 and TNFR2 in both patients with T1DM and those with T2DM. Multivariable regression analyses showed that there were independent associations of FABP4 concentration with body mass index (BMI) and estimated glomerular filtration rate (eGFR) after adjustment for age and sex in both patients with T1DM and those with T2DM. FABP4 concentration was independently associated with circulating levels of TNFR1 and TNFR2 after adjustment for the confounders in patients with T2DM but not in those with T1DM. Similarly, levels of TNFR1 and TNFR2 were independently associated with FABP4 concentration after adjustment for age, sex, systolic blood pressure, duration of DM and levels of eGFR, high-density lipoprotein cholesterol, and C-reactive protein in patients with T2DM but not in those with T1DM. Conclusion: FABP4 concentration is independently associated with levels of TNFRs in patients with DM, but the association is more evident in patients with T2DM than in those with T1DM.

Marine pearls from Pinctada fucata martensii are valuable for decoration, skin care, and health care, but their quality and yield can be threatened by diseases, pearl nuclei transplantation rejection, and other external factors. The present study investigated the immune function of the tumor necrosis factor receptor (TNFR) gene in P. f. martensii to better understand immune responses related to nuclei rejection. The PmTNFR16 cDNA sequence obtained from cloning was 1675 bp in length and encoded 406 amino acid residues. Sequence analysis revealed that PmTNFR16 comprises one signal peptide domain, four TNFR domains, one transmembrane domain, and one death domain. Expression analysis revealed PmTNFR16 was highly expressed in the mantle and significantly up-regulated after exogenous stimuli. The effects of PmTNFR16 on immune effector molecules within the NF-κB signaling pathway were further explored using RNA interference probes for PmTNFR16. Pre-treatment of pearl oysters with dsRNA-PmTNFR before nucleus-insertion resulted in decreased expression of PmTNFR16 by 53 % at 24 h (p

Introduction: Large-scale clinical trials of sodium-glucose cotransporter 2 inhibitors (SGLT2i) demonstrate proteinuria-reducing effects in diabetic kidney disease, even after treatment with renin–angiotensin inhibitors. The precise mechanism for this favorable effect remains unclear. This prospective open-label single-arm study investigated factors associated with a reduction in proteinuria after SGLT2i administration. Methods: Patients with type 2 diabetes (T2DM) who had glycated hemoglobin (HbA1c) levels ≥ 6.5% despite dietary and/or oral hypoglycemic monotherapy were recruited and administered the recommended daily dose of SGLT2i for 4 months. Dual primary outcomes were changes in the urine albumin-to-creatinine ratio (uACR) and urine liver-type fatty acid-binding protein (L-FABP)-to-creatinine ratio (uL-FABPCR) at month 4 from baseline. Changes in kidney injury, inflammation, and oxidative stress biomarkers were investigated as secondary endpoints to examine the effects of this treatment on the kidney. The correlation between renal outcomes and clinical indicators, including circulating tumor necrosis factor receptors (TNFR) 1 and 2, was evaluated using univariate and multivariate analyses. Results: Participants (n = 123) had a mean age of 64.1 years (SD 13.4), with 50.4% being male. The median BMI was 25.8 kg/m2 (interquartile range (IQR) 23.1–28.9), and the median HbA1c level was 7.3% (IQR 6.9–8.3). After SGLT2i administration, the uACR declined from 19.2 mg/gCr (IQR 7.1–48.7) to 13.3 mg/gCr (IQR 7.5–31.6), whereas the uL-FABPCR was not influenced. In univariate analysis, the change in log-transformed uACR due to SGLT2i administration showed a positive correlation with the change in serum TNFR1 level (R = 0.244, p < 0.01). Multivariate regression analysis, including confounding factors, showed that the changes in serum TNFR1 level were independently associated with the changes in the log-transformed uACR (independent t = 2.102, p < 0.05). Conclusion: After the 4-month SGLT2i administration, decreased albuminuria level was associated with decreased serum TNFR level in patients with T2DM. Trial Registration Number: UMIN000031947. Plain Language Summary: Previous studies have demonstrated the synergistic proteinuria-reducing effect of sodium-glucose cotransporter 2 inhibitors (SGLT2i) in combination therapy with renin–angiotensin system blockers; however, the underlying mechanisms of this effect are poorly understood. This study was based on our hypothesis that the proteinuria-reducing effect is associated with the anti-inflammatory effects of SGLT2i beyond the effect on glycemic control. In total, 123 patients with type 2 diabetes mellitus (T2DM) were administered the recommended daily dose of SGLT2i for 4 months. Dual primary outcomes were changes in the urine albumin-to-creatinine ratio (uACR) and urine liver-type fatty acid-binding protein (L-FABP)-to-creatinine ratio (uL-FABPCR) as markers of glomerular and proximal tubular damage at 4 months from the baseline. Secondary outcomes included changes in kidney injury biomarkers, inflammation, and oxidative stress to examine the effects of treatment on the kidneys. The correlation between renal outcomes and clinical indicators, including circulating tumor necrosis factor receptors (TNFR) 1 and 2, was evaluated using univariate and multivariate analyses. We found that administration of SGLT2i decreased the urine albumin-to-creatinine ratio but did not affect the urine liver-type fatty acid-binding protein-to-creatinine ratio. Further, SGLT2i may exert a proteinuria-reducing effect dependent on the anti-inflammatory effect in patients with T2DM. The inflammation-reducing and renoprotective mechanisms of SGLT2i remain to be fully clarified, but this study provides novel evidence regarding the mechanism. The study findings can help in developing anti-inflammatory agents for metabolic diseases. [ABSTRACT FROM AUTHOR]

The goal of our study was to evaluate the potential role of sTNF-RI as a biomarker of renal involvement in SLE patients and active SLE. The study sample consisted of two cohorts. The discovery cohort included 16 SLE patients without renal involvement (non-LN), 60 lupus nephritis (LN) patients and 21 healthy controls (HCs) and the replication cohort included 18 SLE non-LN patients, 116 LN patients and 36 HCs. The sTNF-RI levels differed significantly in the discovery cohort. The plasma sTNF-RI levels were higher in LN patients than in non-LN patients (p =.009) and HCs (p = 4 × 10−6). Plasma sTNF-RI levels were significantly higher in non-LN patients than in HCs (p =.03). The finding was confirmed in independent replication cohort (LNs vs. non-LN, p = 4.053 × 10−7; LNs vs. HCs, p = 2.395 × 10−18; non-LN vs. HCs, p = 2.51 × 10−4). The plasma sTNF-RI levels were associated with disease activity, renal function in SLE patients and urine protein in LN patients. The multivariate analysis revealed that high sTNF-RI was an independent risk factor for renal involvement. The multivariate logistic regression results suggested that high TNF-RI, high systolic blood pressure, high serum creatinine, low C4 and positive anti-dsDNA were independent risks of active SLE patients. A nomogram was constructed based on the results of multivariate logistic regression analysis and it was practical in predicting the risk of the active SLE patients. Immunohistochemistry suggested that the expression of TNF-RI in the kidney was increased. Plasma sTNF-RI might be a good biomarker of renal involvement and disease activity in SLE patients. [ABSTRACT FROM AUTHOR]

The amino acids arginine (Arg), asymmetric (ADMA) and symmetric dimethylarginine (SDMA) are related to nitric oxide (NO) metabolism and potential markers of two different disease entities: cardiovascular disease such as atherosclerosis and systemic inflammation in critically ill patients with sepsis. Although very different in their pathophysiological genesis, both entities involve the functional integrity of blood vessels. In this context, large population-based data associating NO metabolites with proinflammatory markers, e.g., white blood cell count (WBC), high-sensitivity C-reactive protein (hsCRP), and fibrinogen, or cytokines are sparse. We investigated the association of Arg, ADMA and SDMA with WBC, hsCRP, and fibrinogen in 3556 participants of the Study of Health in Pomerania (SHIP)-TREND study. Furthermore, in a subcohort of 456 subjects, 31 inflammatory markers and cytokines were analyzed. We identified Arg and SDMA to be positively associated with hsCRP (β coefficient 0.010, standard error (SE) 0.002 and 0.298, 0.137, respectively) as well as fibrinogen (β 5.23 × 10−3, SE 4.75 × 10−4 and 0.083, 0.031, respectively). ADMA was not associated with WBC, hsCRP, or fibrinogen. Furthermore, in the subcohort, Arg was inversely related to a proliferation-inducing ligand (APRIL). SDMA was positively associated with osteocalcin, tumor necrosis factor receptor 1 and 2, and soluble cluster of differentiation 30. Our findings provide new insights into the involvement of Arg, ADMA, and SDMA in subclinical inflammation in the general population. [ABSTRACT FROM AUTHOR]

AbstractObjectives The goal of our study was to evaluate the potential role of sTNF-RI as a biomarker of renal involvement in SLE patients and active SLE.Methods The study sample consisted of two cohorts. The discovery cohort included 16 SLE patients without renal involvement (non-LN), 60 lupus nephritis (LN) patients and 21 healthy controls (HCs) and the replication cohort included 18 SLE non-LN patients, 116 LN patients and 36 HCs.Results The sTNF-RI levels differed significantly in the discovery cohort. The plasma sTNF-RI levels were higher in LN patients than in non-LN patients (p = .009) and HCs (p = 4 × 10−6). Plasma sTNF-RI levels were significantly higher in non-LN patients than in HCs (p = .03). The finding was confirmed in independent replication cohort (LNs vs. non-LN, p = 4.053 × 10−7; LNs vs. HCs, p = 2.395 × 10−18; non-LN vs. HCs, p = 2.51 × 10−4). The plasma sTNF-RI levels were associated with disease activity, renal function in SLE patients and urine protein in LN patients. The multivariate analysis revealed that high sTNF-RI was an independent risk factor for renal involvement. The multivariate logistic regression results suggested that high TNF-RI, high systolic blood pressure, high serum creatinine, low C4 and positive anti-dsDNA were independent risks of active SLE patients. A nomogram was constructed based on the results of multivariate logistic regression analysis and it was practical in predicting the risk of the active SLE patients. Immunohistochemistry suggested that the expression of TNF-RI in the kidney was increased.Conclusions Plasma sTNF-RI might be a good biomarker of renal involvement and disease activity in SLE patients.

TNFα blocking agents were the first-in-class biologic drugs used for the treatment of autoimmune disease. Paradoxically, however, exacerbation of autoimmunity was observed in some patients. TNFα is a pleiotropic cytokine that has both proinflammatory and regulatory effects on CD4+ T cells and can influence the adaptive immune response against autoantigens. Here, we critically appraise the literature and discuss the intricacies of TNFα signaling that may explain the controversial findings of previous studies. The pleiotropism of TNFα is based in part on the existence of two biologically active forms of TNFα, soluble and membrane-bound, with different affinities for two distinct TNF receptors, TNFR1 and TNFR2, leading to activation of diverse downstream molecular pathways involved in cell fate decisions and immune function. Distinct membrane expression patterns of TNF receptors by CD4+ T cell subsets and their preferential binding of distinct forms of TNFα produced by a diverse pool of cellular sources during different stages of an immune response are important determinants of the differential outcomes of TNFα-TNF receptor signaling. Targeted manipulation of TNFα-TNF receptor signaling on select CD4+ T cell subsets may offer specific therapeutic interventions to dampen inflammation while fortifying immune regulation for the treatment of autoimmune diseases. [ABSTRACT FROM AUTHOR]

TNFa blocking agents were the first-in-class biologic drugs used for the treatment of autoimmune disease. Paradoxically, however, exacerbation of autoimmunity was observed in some patients. TNFa is a pleiotropic cytokine that has both proinflammatory and regulatory effects on CD4+ T cells and can influence the adaptive immune response against autoantigens. Here, we critically appraise the literature and discuss the intricacies of TNFa signaling that may explain the controversial findings of previous studies. The pleiotropism of TNFa is based in part on the existence of two biologically active forms of TNFa, soluble and membrane-bound, with different affinities for two distinct TNF receptors, TNFR1 and TNFR2, leading to activation of diverse downstream molecular pathways involved in cell fate decisions and immune function. Distinct membrane expression patterns of TNF receptors by CD4+ T cell subsets and their preferential binding of distinct forms of TNFα produced by a diverse pool of cellular sources during different stages of an immune response are important determinants of the differential outcomes of TNFa-TNF receptor signaling. Targeted manipulation of TNFa-TNF receptor signaling on select CD4+ T cell subsets may offer specific therapeutic interventions to dampen inflammation while fortifying immune regulation for the treatment of autoimmune diseases.

Förster resonance energy transfer (FRET) is the direct energy exchange between two‐component fluorescent molecules. FRET methods utilize chemically linked molecules or unlinked fluorescent molecules such as fluoresscent protein–protein interactions. FRET is therefore a powerful indicator of molecular proximity, but standardized determination of FRET efficiency is challenged when investigating natural (chemically unlinked) interactions. In this paper, we have examined the interactions of tumor necrosis factor receptor‐1 (TNFR1) molecules expressed as recombinant C‐terminal fusion proteins of cyan, yellow, or red fluorescent protein (‐CFP, ‐YFP, or ‐RFP) to evaluate two‐molecule chemically unlinked FRET by flow cytometry. We demonstrate three independent FRET pairs of TNFR1 CFP→YFP (FRET‐1), YFP→RFP (FRET‐2) and CFP→RFP (FRET‐3), by comparing TNFR1+TNFR1 with non‐interacting TNFR1+CD27 proteins, on both LSR‐II and Fortessa X‐20 cytometers. We describe genuine FRET activities reflecting TNFR1 homotypic interactions. The FRET events can be visualized during sample acquisition via the use of "spiked" FRET donor cells, together with TNFR1+TNFR1 co‐transfected cells, as FRET channel mean fluorescence intensity (MFI) overlays. FRET events can also be indicated by comparing concatenated files of cells expressing either FRET positive events (TNFR1+TNFR1) or FRET negative events (TNFR1+CD27) to generate single‐cell scatter plots showing loss of FRET donor brightness. Robust determination of FRET efficiency is then confirmed at the single‐cell level by applying matrix calculations based on the measurements of FRET, using donor, acceptor, and FRET fluorescent intensities (I), detector channel emission coefficient (S), fluorescent protein extinction coefficients (ε) and the α factor. In this TNFR1‐based system the mean CFP→YFP FRET‐1 efficiency is 0.43 (LSR‐II) and 0.41 (Fortessa X‐20), the mean YFP→RFP FRET‐2 efficiency is 0.30 (LSR‐II) and 0.29 (Fortessa X‐20), and the mean CFP→RFP FRET‐3 efficiency is 0.56 (LSR‐II) and 0.54 (Fortessa X‐20). This study also embraces multi‐dimensional clustering using t‐SNE, Fit‐SNE, UMAP, Tri‐Map and PaCMAP to further demonstrate FRET. These approaches establish a robust system for standardized detection of chemically unlinked TNFR1 homotypic interactions with three individual FRET pairs. [ABSTRACT FROM AUTHOR]

Background and Objective: CD137 is a member of the TNF-Receptor family. TNF-alpha antagonists have therapeutic effect in active psoriasis. In this study, the relative frequency of CD137 expression was investigated in the inflammatory cells of psoriasis lesions for the first time. Methods: The specimens were obtained from pathology department of Al-Zahra hospital from 2007 to 2016, from paraffin-embedded skin biopsies. A number of 64 psoriasis skin lesions specimens and 34 normal skin specimens were rechecked for the diagnosis. Then, the immunohistochemical staining for CD137, CD4, and CD8 was performed. Results: CD137 expression of dermal inflammatory cells in psoriasis lesion was 11.19±5.5%. Although, in normal skin biopsied, CD137 expression was observed in 1.3±3.03% of the inflammatory cells. (p =0.001). The relative frequency of the CD137 positive inflammatory cells was significantly more in epidermis versus dermis (epidermis: 31.1%±12.8, dermis 11.1%±5.5). There was no remarkable relation between the CD137 expression rate and the CD4: CD8 ratio. Conclusion: CD137 as a TNF-alpha receptor has a significant role in pathogenesis of the psoriasis lesions. Therefore, CD137 antagonists can be considered as a novel target for the treatment of incurable psoriasis patients.

Abstract Aims/Introduction Increased concentrations of serum tumor necrosis factor (TNF) receptors (TNFRs; TNFR1 and TNFR2) are positively associated with the urinary albumin‐to‐creatinine ratio (ACR), and negatively associated with the estimated glomerular filtration rate (eGFR) in patients with type 2 diabetes. However, the mechanism underlying this increase and the relationship between TNFRs in serum, and urine and kidney measures (ACR and eGFR) are unclear. Materials and Methods This was a cross‐sectional study that included 499 patients with type 2 diabetes and eGFR ≥60 mL/min/1.73 m2. The concentrations of TNFRs in serum and urine, and their respective fractional excretion, were measured. Results Serum and urinary TNFR levels were positively associated with the ACR, and negatively associated with the eGFR. The fractional excretion of TNFRs did not differ between patients with an eGFR ≥90 and those with an eGFR 60–89 mL/min/1.73 m2, and also did not correlate with eGFR. After adjustment for relevant covariates, the serum TNFRs were associated with a lower eGFR (60–89 mL/min/1.73 m2) and an increased ACR (≥30 mg/gCr), but urinary TNFRs were associated with an increased ACR (≥30 mg/gCr) alone, in the multivariate logistic model. Conclusions The pattern of fractional excretion TNFRs showed that an increase in serum TNFRs might result from their increased systemic production, including in the kidney, rather than being a simple reflection of GFR decline. Kidney measures appear to be strongly associated with serum TNFRs rather than urinary TNFRs in patients with type 2 diabetes and normal renal function.

Background: Fatty acid-binding protein 4 (FABP4) is an adipokine that plays significant roles in the development of insulin resistance and atherosclerosis. High levels of soluble tumor necrosis factor receptors (TNFRs) including TNFR1 and TNFR2 are associated with renal dysfunction and increased mortality in patients with diabetes mellitus (DM). However, the association between circulating levels of FABP4 and TNFRs remains unclear., Methods: We investigated the associations of FABP4 with TNFRs and metabolic markers in Japanese patients with type 1 DM (T1DM, n = 76, men/women: 31/45) and type 2 DM (T2DM, n = 575, men/women: 312/263)., Results: FABP4 concentration was positively correlated with levels of TNFR1 and TNFR2 in both patients with T1DM and those with T2DM. Multivariable regression analyses showed that there were independent associations of FABP4 concentration with body mass index (BMI) and estimated glomerular filtration rate (eGFR) after adjustment for age and sex in both patients with T1DM and those with T2DM. FABP4 concentration was independently associated with circulating levels of TNFR1 and TNFR2 after adjustment for the confounders in patients with T2DM but not in those with T1DM. Similarly, levels of TNFR1 and TNFR2 were independently associated with FABP4 concentration after adjustment for age, sex, systolic blood pressure, duration of DM and levels of eGFR, high-density lipoprotein cholesterol, and C-reactive protein in patients with T2DM but not in those with T1DM., Conclusion: FABP4 concentration is independently associated with levels of TNFRs in patients with DM, but the association is more evident in patients with T2DM than in those with T1DM.

An immune tolerant tumor microenvironment promotes immune evasion of lung cancer. Agents that antagonize immune tolerance will thus aid the fight against this devastating disease. Members of the tumor necrosis factor receptor (TNFR) family modulate the magnitude, duration and phenotype of immune responsiveness to antigens. Among these, GITR expressed on immune cells functions as a key regulator in inflammatory and immune responses. Here, we evaluate the GITR agonistic antibody (DTA-1) as a mono-therapy and in combination with therapeutic vaccination in murine lung cancer models. We found that DTA-1 treatment of tumor-bearing mice increased: (i) the frequency and activation of intratumoral natural killer (NK) cells and T lymphocytes, (ii) the antigen presenting cell (APC) activity in the tumor, and (iii) systemic T-cell specific tumor cell cytolysis. DTA-1 treatment enhanced tumor cell apoptosis as quantified by cleaved caspase-3 staining in the tumors. DTA-1 treatment increased expression of IFNγ, TNFα and IL-12 but reduced IL-10 levels in tumors. Furthermore, increased anti-angiogenic chemokines corresponding with decreased pro-angiogenic chemokine levels correlated with reduced expression of the endothelial cell marker Meca 32 in the tumors of DTA-1 treated mice. In accordance, there was reduced tumor growth (8-fold by weight) in the DTA-1 treatment group. NK cell depletion markedly inhibited the antitumor response elicited by DTA-1. DTA-1 combined with therapeutic vaccination caused tumor rejection in 38% of mice and a 20-fold reduction in tumor burden in the remaining mice relative to control. Mice that rejected tumors following therapy developed immunological memory against subsequent re-challenge. Our data demonstrates GITR agonist antibody activated NK cell and T lymphocyte activity, and enhanced therapeutic vaccination responses against lung cancer.

β-1,4 Galactosyltransferase V (β-1,4-GalT V) belongs to the β-1,4 galactosyltransferase family, which modifies proteins and plays a vital role in biological function. Our previous study revealed that β-1,4-GalT V was expressed in the cortex and hippocampus and participated in the recovery of spatial learning and memory in rats with traumatic brain injury. However, the expression of β-1,4-GalT V in microglia, resident immune cells in the central nervous system, and its impact on microglia in resting and lipopolysaccharide-triggered activated stages are elusive. In this study, we clarified that β-1,4-GalT V expresses in microglia, and it regulates microglial migration, proliferation, and release of the inflammatory factors. We also observed that β-1,4-GalT V affects the expression level of tumor necrosis factor receptor (TNFR)2 instead of TNFR1. These results strongly support the fact that β-1,4-GalT V is involved in microglial function. [ABSTRACT FROM AUTHOR]

Background & Objective: CD137 is a member of the TNF-Receptor family. TNFalpha antagonists have therapeutic effect in active psoriasis. In this study, the relative frequency of CD137 expression was investigated in the inflammatory cells of psoriasis lesions for the first time. Methods: The specimens were obtained from pathology department of Al-Zahra hospital from paraffin-embedded skin specimens collected from 2007 till 2016. A total number of f 64 psoriasis skin specimens and 34 normal skin specimens were reviewed for the diagnosis. Then, the immunohistochemical staining for CD137, CD4, and CD8 was performed. Results: CD137 expression of dermal inflammatory cells in psoriasis lesions was 11.19±5.5%. Although, in normal skin tissues, CD137 expression was observed in 1.3±3.03% of the inflammatory cells. (P=0.001). The relative frequency of the CD137 positive inflammatory cells was significantly higher in the epidermis compared to dermis (epidermis: 31.1%±12.8, dermis 11.1%±5.5). There was no remarkable relation between the CD137 expression rate and the CD4: CD8 ratio. Conclusion: CD137 as a TNF-alpha receptor has a significant role in pathogenesis of the psoriasis lesions. Therefore, CD137 antagonists can be considered as a novel target for the treatment of incurable psoriasis patients. [ABSTRACT FROM AUTHOR]

Aims/Introduction: Increased concentrations of serum tumor necrosis factor (TNF) receptors (TNFRs; TNFR1 and TNFR2) are positively associated with the urinary albumin‐to‐creatinine ratio (ACR), and negatively associated with the estimated glomerular filtration rate (eGFR) in patients with type 2 diabetes. However, the mechanism underlying this increase and the relationship between TNFRs in serum, and urine and kidney measures (ACR and eGFR) are unclear. Materials and Methods: This was a cross‐sectional study that included 499 patients with type 2 diabetes and eGFR ≥60 mL/min/1.73 m2. The concentrations of TNFRs in serum and urine, and their respective fractional excretion, were measured. Results: Serum and urinary TNFR levels were positively associated with the ACR, and negatively associated with the eGFR. The fractional excretion of TNFRs did not differ between patients with an eGFR ≥90 and those with an eGFR 60–89 mL/min/1.73 m2, and also did not correlate with eGFR. After adjustment for relevant covariates, the serum TNFRs were associated with a lower eGFR (60–89 mL/min/1.73 m2) and an increased ACR (≥30 mg/gCr), but urinary TNFRs were associated with an increased ACR (≥30 mg/gCr) alone, in the multivariate logistic model. Conclusions: The pattern of fractional excretion TNFRs showed that an increase in serum TNFRs might result from their increased systemic production, including in the kidney, rather than being a simple reflection of GFR decline. Kidney measures appear to be strongly associated with serum TNFRs rather than urinary TNFRs in patients with type 2 diabetes and normal renal function. [ABSTRACT FROM AUTHOR]

Background: Ligation of glucocorticoid‐induced tumor necrosis factor receptor (GITR) decreases regulatory T cell–mediated suppression and enhances T‐cell proliferation, effector function, and survival. MK‐1248 is a humanized immunoglobulin G4 anti‐GITR monoclonal antibody agonist. Methods: In patients with advanced solid tumors, MK‐1248 (starting dose, 0.12 mg) was tested alone and with pembrolizumab (200 mg) according to a 3 + 3 dose escalation design (ClinicalTrials.gov identifier NCT02553499); both treatments were administered intravenously every 3 weeks for ≤4 and ≤35 cycles, respectively. The safety and tolerability, maximum tolerated dose, and pharmacokinetics/pharmacodynamics were explored. Results: Twenty patients received MK‐1248 monotherapy; 17 received combination therapy. The most frequent tumor types were colorectal cancer (n = 8), melanoma (n = 6), and renal cell carcinoma (n = 4). MK‐1248 was generally well tolerated at the maximum tested doses of 170 (monotherapy) and 60 mg (combination). No dose‐limiting toxicities (DLTs) or treatment‐related deaths occurred. Adverse events (AEs) occurred in 36 of the 37 patients (97%); the most common were vomiting (n = 13 [35%]), anemia (n = 10 [27%]), and decreased appetite (n = 10 [27%]). Grade 3 to 5 AEs occurred in 19 of the 37 patients (51%). Treatment‐related AEs occurred in 18 of the 37 patients (49%): 9 of the 20 patients (45%) on monotherapy and 9 of the 17 patients (53%) on combination therapy. Among the 17 patients receiving combination therapy, 1 achieved a complete response and 2 achieved a partial response, for an objective response rate of 18%; no patients achieved an objective response with monotherapy. The disease control rate (stable disease or better) was 15% with monotherapy and 41% with combination therapy. Conclusions: MK‐1248 was generally well tolerated at doses up to 170 (monotherapy) and 60 mg (combination), with no DLTs or treatment‐related deaths. Combination therapy provided limited antitumor responses. MK‐1248 has a manageable safety profile and limited antitumor activity when it is combined with pembrolizumab in patients with advanced solid tumors. The role of combination therapy with a glucocorticoid‐induced tumor necrosis factor receptor agonist and an immune checkpoint inhibitor still needs to be elucidated. [ABSTRACT FROM AUTHOR]

Objective: To explore the effect of imbalance of tumor necrosis factor receptor related factor-6 (TRAF-6), interleukin 1 receptor associated kinase-1 (IRAK-1) and neutrophil alkaline phosphatase-3 (NALP3) in patients with gouty arthritis. Methods: The retrospective experiment was conducted on 105 patients with gouty arthritis admitted to our hospital (47 patients with acute onset and 58 patients with remission, namely group A and group B); meanwhile, another 61 healthy volunteers were selected for control, namely group C. The enrolling of the three groups was dated from May 2017 to May 2018, and TRAF-6, IRAK-1 and NALP3 of all subjects were tested through real-time fluorescence quantification (RT-PCR), and the correlation between the three inflammatory factors and gouty arthritis was compared. Results: 1) Through treatment, ESR, BUA and total addiment in group A and B were higher than those in group C, among which the three indicators in group A were higher than those in group B (P<0.05), while CRP was lower than that of group C, and the two indicators in group A were lower than those in group B (P<0.05). 2) There was no significant difference in the relative expression of TRAF-6 mRNA between group A and group B before treatment (P>0.05), significantly lower than group C (P<0.05); the above indicators of group A and group B were improved to some extent after treatment, but group A was still lower than group B (P<0.05), and the degree of improvement of group A was also lower than that of group C (P<0.05), while the degree of improvement of group B was not significantly different from that of group C (P>0.05). 3) The relative expression level of IRAK-1mRNA in group A and group B before treatment showed no significant difference (P>0.05), but was also lower than that in group C (P<0.05). The relative expression level of IRAK-1mRNA in group A and group B increased to some extent after treatment, with group A significantly lower than group C (P<0.05), and group B showed no significant difference compared with group C (P> 0.05). 4) The relative expression level of NALP-3 mRNA in group A and group B showed no significant difference (P>0.05) before treatment, significantly higher than that in group C (P <0.05); the relative expression of NALP-3 mRNA in group A was not significantly decreased (P>0.05) after treatment, while that in group B was significantly decreased after treatment (P < 0.05), indicating significant different compared with group A and group C. 5) There was no correlation between) TRAF-6, ESR, CRP and total addiment (P>0.05); IRAK-1 was negatively correlated with CRP, BUA and total addiment (P<0.05); NALP-3 was negatively correlated with ESR and CRP (P<0.05). Conclusion: TRAF-6, IRAK-1 and NALP-3 are all under abnormal expression in the developing of new gouty arthritis, acting as important participants in promoting the occurrence, development and outcome of illness states, so the intervening measures should be taken. [ABSTRACT FROM AUTHOR]

AIM: To examine the association of genetic polymorphisms (-308)G/A TNFα, (+250)A/G Ltα, (+36)A/G TNFR1, (+1663)A/G TNFR2 with the development of primary open angle glaucoma (POAG) among people in Central Russia. METHODS: The study sample included 443 individuals, of which 252 patients with POAG and 191 individuals in the control group. Genotyping of (-308)G/A TNFα, (+250)A/G Ltα, (+36)A/G TNFR1, (+1663)A/G TNFR2 was performed using polymerase chain reaction. The distribution of alleles and genotypes of the studied DNA markers in the groups was examined by 2×2 contingency tables and χ2 with the Yates’s correction for continuity and odds ratios (OR) with 95% confidence intervals (CI). RESULTS: Allele (-308)G TNFα (Р=0.01, OR=1.78, 95%CI 1.12-2.85) was identified as a risk factor for POAG. Homozygotes (-308) AA TNFα are at a lowest risk for development of the disease (Р=0.01, OR=0.0005). The following combination of genetic variants of cytokines were associated with a reduced risk of POAG: (+1663)A TNFR2 and (+250)G Ltα (OR=0.34) CONCLUSION: Genetic polymorphisms (-308)G/A TNFα, (+250)A/G Ltα, (+1663)A/G TNFR2 associated with the development of POAG in the population of Central Russia.

In this study, an echinoderm tumor necrosis factor receptor named HLTNFR-16 was first cloned from the tropical sea cucumber Holothuria leucospilota. The full-length cDNA of HLTNFR-16 is 3675 bp in size, containing a 415 bp 5′-untranslated region (UTR), a 2024 bp 3′-UTR and a 1236 bp open reading frame (ORF) encoding a protein of 411 amino acids with a deduced molecular weight of 45.63 kDa. The HLTNFR-16 protein contains a signal peptide, four TNFR domains (the last three were identified as extracellular cysteine-rich domains), a transmembrane region and a death domain. Phylogenetic analysis showed that HLTNFR-16 was clustered into a clade with TNFR-16s in other species, indicating that this echinoderm TNFR may be a new member of the TNFR-16 subfamily. The results of TUNEL assay showed that the over expression of HLTNFR-16 could induce apoptosis in HEK293T cells. When HLTNFR-16 was silenced by siRNA, the apoptosis of sea cucumber coelomocytes induced by inactivated Vibrio harveyi was suppressed significantly, indicating that HLTNFR-16 is important for apoptosis induction. Additionally, luciferase reporter assay exhibited that the over-expressed HLTNFR-16 in HEK293T cells could activate the transcription factors nuclear factor-κB (NF-κB) and activator protein-1 (AP-1). Moreover, the secretion of proinflammatory cytokines interleukin (IL)-1β, IL-6 and IL-18 in HEK293T cells was increased by the over-expression of HLTNFR-16. This study provides evidences for the potential roles of sea cucumber TNFR in the innate immunity. • We identified a TNFR-16 from the tropical sea cucumber Holothuria leucospilota. • Over-expression of HLTNFR-16 could induce apoptosis in HEK293T cells. • The silencing of HLTNFR-16 could significantly suppress the apoptosis of sea cucumber coelomocytes induced by inactivated Vibrio harveyi. • Over-expressed HLTNFR-16 could activate the transcription factors nuclear factor-κB and activator protein-1. • Over-expressed HLTNFR-16 could induce the secretion of proinflammatory cytokines. [ABSTRACT FROM AUTHOR]

Objective: To explore the effect of Linagliptin on tumor necrosis factor receptor and monocyte chemoattractant protein-1 in patients with diabetic nephropathy. Methods: A total of 98 patients with diabetic nephropathy admitted to the Hospital from January 2017 to September 2018 were enrolled. The patients were divided into two groups according to the random double-blind method, with 49 cases in each group. The control group was treated with Metformin, whereas the experimental group was treated with Linagliptin plus Metformin. After 3 months of continuous treatment, the renal function [urinary albumin excretion rate, 24 h urine protein quantitation and serum creatinine], glycolipids metabolic levels [glycated hemoglobin, fasting blood glucose, total cholesterol and triglycerides], monocyte chemoattractant protein-1, tumor necrosis factor receptor, high-sensitivity C-reactive protein, and adverse reactions were compared between the two groups. Results: After 3 months of treatment, the levels of UAER, 24 h Upor and Scr in the experimental group were shown to be lower than those in the control group, and the difference was statistically significant. After 3 months of treatment, the levels of HbA1c, FPG, TC and TG in the experimental group were shown to be lower than the control group, and the difference was statistically significant. After 3 months of treatment, the levels of MCP-1, sTNFR1 and hs-CRP in the experimental group were lower than those in the control group, and the difference was statistically significant. There was no significant difference in incidence of adverse reactions between the two groups. Conclusion: For patients with diabetic nephropathy, Linagliptin is with higher safety, which can help improve their glycolipids metabolic levels and renal function, reduce the inflammatory response and the levels of MCP-1 and sTNFR1, and yet incur fewer adverse reactions. [ABSTRACT FROM AUTHOR]

Objective To study the intervention effect of emodin on Con A-induced acute liver injury in mice and the underlying mechanism. Methods The male Balb/C mice were randomly divided into the control group, model group, and emodin group. Emodin(25 mg/kg) was administered orally, and Con A(20 mg/kg) was given through intravenous injection at 2 h after emodin administration in the emodin group. In place of emodin, CMC-Na was used in the model group, and mice in the control group were not treated.After 10 h of Con A injection, the blood was collected from the retro-orbital sinus in mice and the serum levels of ALT and AST were measured. Hepatic sections were aseptically obtained for histopathological examination to assess the degree of liver damage. The real-time fluorescent quantitative PCR and Western blotting were employed to evaluate the expression of glucocorticoid-induced tumor necrosis factor receptor(GITR) mRNA and its ligand GITRL mRNA.Results The serum ALT and AST levels in the model group were significantly higher than those in the control group, while those in the emodin group were lower than those in the model group(all P<0.01). Compared with the control group, there were more significant necrosis foci in the liver tissues of the model group, while the liver necrosis was significantly reduced in the emodin group. The relative expression levels of GITR and GITRL mRNA in the model group were higher than those in the control group, while the relative expression levels of GITR and GITRL mRNA in the emodin group were lower than those in the model group(all P<0.05). The relative expression levels of GITR and GITRL protein in the model group were higher than those in the control group, while the relative expression levels of GITR and GITRL protein in the emodin group were lower than those in the model group(all P<0.05). Conclusion Emodin alleviates Con A-induced liver damage by inhibiting GITR/GITRL pathway. [ABSTRACT FROM AUTHOR]

Abstract The interaction between TNF-α with TNFR1 triggers important signaling pathways inducing diverse cellular phenomena including inflammation, apoptosis, etc., and is involved in the pathogenesis and progression of numerous autoimmune diseases. The extracellular domain (ECD) of TNFR has been successfully used to clinically treat such TNF-associated diseases. However, large-scale production of these biological material via eukaryotic cell expression systems is usually costly owing to the culture medium and complicated growth conditions. This study aimed to extract pure soluble human TNFR1-ECD and investigate its biological activity, using a prokaryotic expression system. Recombinant vector pMCSG7-TNFR1-ECD was constructed via ligation-independent cloning. The His-tag fusion protein was expressed in E. coli and localized in inclusion bodies. Recombinant TNFR1-ECD was refolded and purified via nickel-affinity chromatography, tag cleavage, followed by cation-exchange chromatography or size-exclusion chromatography. A purity of over 95% and a yield of 9.3 mg protein per liter of bacterial culture media was obtained. The purified protein showed significant affinity of 2.15 nM towards human TNF-α and inhibited TNF-α-mediated cytotoxicity in L929 cells, with an ED 50 of 0.10 μg/ml. It formed a self-associated oligomer with a K D of 1.15 μM, detected via microscale thermophoresis. We thus established a highly efficient approach to construct, express, and purify the recombinant protein of human TNFR1-ECD from a prokaryotic system. The antagonistic bioactivities in vitro indicate this protein as a prospective molecules for drug research against autoimmune diseases characterized by TNF-α overexpression. Highlights • A highly efficient approach was established to obtain the extracellular domain of human TNFR1 from prokaryotic system. • The recombinant protein showed high purity and excellent biological function. • NanoDSF was used to showed the high thermal stability of the recombinant TNFR1 ectodomain. • MST was first used to demonstrate the oligomerization of the recombinant TNFR1 ectodomain. [ABSTRACT FROM AUTHOR]

Oncogenic microRNA-21 (miR-21/miRNA-21) is a stable inhibitor of gene expression that is often upregulated in cervical cancer, a disease that affects the health of women and tends to transform and spread. Previous studies investigating miR-21 in biopsies and cells from cervical cancer patients have identified that miR-21 binds target mRNAs in signaling pathways or long non-coding RNAs (lncRNA). Furthermore, studies have elucidated the molecular mechanisms of two tumor necrosis factor α (TNF-α) signaling pathways that promote cell proliferation and inhibit cell apoptosis. miR-21 inhibits the TNF receptor 1 (TNFR1) signaling pathway and activates the TNFR2 signaling pathway. Moreover, miR-21 enhances cervical cancer cell proliferation by influencing the protein kinase B/mammalian target of rapamycin and RAS p21 protein activator 1 signaling pathways. The present review discusses the evidence that miR-21 may impact cervical cancer through inhibiting apoptosis and enhancing proliferation, and may therefore be a target for clinical intervention. [ABSTRACT FROM AUTHOR]

Tumor necrosis factor receptor 1 and 2 (TNFR1 and TNFR2) have been found in brain parenchyma of stroke patients, and plasma levels are increased in the acute phase of stroke. We evaluated associations between TNFR1 and TNFR2 plasma levels and stroke severity, infarct size, and functional outcome. Furthermore, we examined cellular expression of TNFR1 and TNFR2 on leukocyte subpopulations to explore the origin of the increased receptor levels. Blood samples were taken from 33 acute ischemic stroke patients and 10 healthy controls. TNFR1 and TNFR2 plasma concentrations were measured and correlated against the Scandinavian Stroke Scale at admission, infarct volume, and the modified Rankin Scale score three months after stroke onset. Classical, intermediate, and non-classical monocytes as well as neutrophils were purified, and cellular expression of TNFR1 and TNFR2 was examined using flow cytometry. TNFR1 and TNFR2 plasma levels were both increased after ischemic stroke, but we found no correlation with patient outcome measurements. Compared to healthy controls, ischemic stroke patients had decreased non-classical monocyte and neutrophil populations expressing TNFR1 and increased neutrophils expressing TNFR2, and decreased non-classical populations co-expressing both TNFR1 and TNFR2. This study supports the hypothesis of an acute immunological response orchestrated by the peripheral immune system following an ischemic stroke. However, the origin of the increased TNFR1 and TNFR2 plasma levels could not be clearly linked to peripheral monocytes or neutrophils. Future studies are needed and will help clarify the potential role as treatment target.

The role of tumor necrosis factor-α (TNF-α) in shaping the tumor microenvironment is ambiguous. Consistent with its uncertain role in melanoma, TNF-α plays a dual role, either acting as a cytotoxic cytokine or favoring a tumorigenic inflammatory microenvironment. TNF-α signals via two cognate receptors, namely TNFR1 (p55) and TNFR2 (p75), which mediate divergent biological activities. Here, we analyzed the impact of TNFR1 deficiency in tumor progression in the B16.F1 melanoma model. Tumors developed in mice lacking TNFR1 (TNFR1 knock-out; KO) were smaller and displayed lower proliferation compared to their wild type (WT) counterpart. Moreover, TNFR1 KO mice showed reduced tumor angiogenesis. Although no evidence of spontaneous metastases was observed, conditioned media obtained from TNFR1 KO tumors increased tumor cell migration. Whereas the analysis of tumor-associated immune cell infiltrates showed similar frequency of total and M2-polarized tumor-associated macrophages (TAMs), the percentage of CD8+ T cells was augmented in TNFR1 KO tumors. Indeed, functional ex vivo assays demonstrated that CD8+ T cells obtained from TNFR1KO mice displayed an increased cytotoxic function. Thus, lack of TNFR1 attenuates melanoma growth by modulating tumor cell proliferation, migration, angiogenesis and CD8+ T cell accumulation and activation, suggesting that interruption of TNF-TNFR1 signaling may contribute to control tumor burden.

Hodgkin lymphoma is a cancer that can be cured using standard chemotherapy with or without radiation. Although it accounts for only 0.6% of all malignancy worldwide, but it usually affects young adults with median age of 38 years. About 60 to 90% cases can be cured depending on its stage and 5 to 10% cases are refractory to the first-line chemotherapy; while 20 to 30% patients experiencing relapse after receiving the first-line chemotherapy. The relapse causes new problem in treatment. A monoclonal antibody-chemotherapy conjugate, Brentuximab vedotin, was approved by Food Drug Association and European Medicine since 2011 dan was approved by European Medicine Agency since 2012 to treat relapsed classical Hodgkin lymphoma and anaplastic large cell lymphoma (ALCL). Brentuximab vedotin has also been known as anti-CD30. CD30 or Ki-1 or TNFRSF8 is a 120-kD glycoprotein, which is a trans-membrane receptor of Hodgkin lymphoma cells. The glycoprotein was first identified in 1982 using monoclonal antibody against Hodgkin lymphoma-derived cell lines. The glycoprotein was then cloned and recognized as a member of tumor necrosis factor receptor (TNFR) superfamily, which has intracellular, transcellular and extracellular domains. The monoclonal antibody obviously does cause a reaction not only with the Reed-Sternberg (RS) cells of Hodgkin lymphoma, but also with a small number of normal lymphocytes subset, which are located at perifollicular zone as well as lymphoid tumor such as anaplastic large cell lymphoma (ALCL) and other non-lymphoid tumor such as embryonic and pancreas carcinoma, undifferentiated nasopharyngeal carcinoma and malignant melanoma. Therefore, CD30 monoclonal antibody alone to confirm the diagnosis of Hodgkin lymphoma is ineffective as it must be used together with other panel of immunohistochemistry antibodies such as cytokeratins, carcinoma embryonic antigen, melanoma-associated antigen and placental alkaline phosphatide. The expression of CD30 molecules in Reed-Sternberg cells of Hodgkin lymphoma has been demonstrated in over 98% of classical Hodgkin lymphoma cases; however, there is a difference in staining intensity among various cases or even in one case.

In this study, we developed Solid Lipid Nanoparticles (SLN-NPs) loaded with Artemisia vulgaris essential oil and coated with folic acid-chitosan (AVEO-SCF-NPs) to enhance drug delivery in biotechnology and pharmaceutical sectors. AVEO-SCF-NPs were synthesized using homogenization and ultra-sonication methods and comprehensively characterized. These nanoparticles exhibited a particle size of 253.67 nm, Polydispersity Index (PDI) of 0.26, zeta potential (ζ-p) of +39.96 mV, encapsulation efficiency (%EE) of 99.0 %, and folic acid binding efficiency (% FB) of 46.25 %. They effectively inhibited MCF-7, HT-29, and PC-3 cancer cells with IC 50 values of 48.87 μg/mL, 88.48 μg/mL, and 121.34 μg/mL, respectively, and demonstrated antibacterial properties against Gram-positive strains. AVEO-SCF-NPs also exhibited scavenging effects on ABTS (IC 50 : 203.83 μg/mL) and DPPH (IC50: 680.86 μg/mL) free radicals and inhibited angiogenesis, as confirmed through CAM and qPCR assays. Furthermore, these nanoparticles induced apoptosis, evidenced by up-regulation of caspase 3 and 9, down-regulation of TNF-α genes, and an increase in SubG1 phase cells. The high loading capacity of SCF-NPs for AVEO, coupled with their multifaceted biological properties, highlights AVEO-SCF-NPs as promising candidates for cancer therapy in the biotechnology and pharmaceutical industries., (© 2024 Wiley-VHCA AG, Zurich, Switzerland.)

Tumor Necrosis Factor-α (TNF-α) is a proinflammatory cytokine implicated in neuronal damage in response to cerebral ischemia. Ischemic preconditioning (IPC) provides neuroprotection against a subsequent severer or longer transient ischemia by ischemic tolerance. Here, we focused on the role of TNF-α in IPC-mediated neuroprotection against neuronal death following a subsequent longer transient cerebral ischemia (TCI). Gerbils used in this study were randomly assigned to eight groups; sham group, TCI operated group, IPC plus (+) sham group, IPC + TCI operated group, sham + etanercept (an inhibitor of TNF-a) group, TCI + etanercept group, IPC + sham + etanercept group, and IPC + TCI + etanercept group. IPC was induced by a 2-min sublethal transient ischemia, which was operated 1 day prior to a longer (5-min) TCI. A significant death of neurons was found in the stratum pyramidale (SP) in the CA1 area (CA1) of the hippocampus 5 days after TCI; however, IPC protected SP neurons from TCI. We found that TNF-α immunoreactivity was significantly increased in CA1 pyramidal neurons in the TCI and IPC + TCI groups compared to the sham group. TNF-R1 expression in CA1 pyramidal neurons of the TCI group was also increased 1 and 2 days after TCI; however, in the IPC + TCI group, TNF-R1 expression was significantly lower than that in the TCI group. On the other hand, we did not detect TNF-R2 immunoreactivity in CA1 pyramidal neurons 1 and 2 days after TCI; meanwhile, in the IPC + TCI group, TNF-R2 expression was significantly increased compared to TNF-R2 expression at 1 and 2 days after TCI. In addition, in this group, TNF-R2 was newly expressed in pericytes, which are important cells in the blood brain barrier, from 1 day after TCI. When we treated etanercept to the IPC + TCI group, IPC-induced neuroprotection was significantly weakened. In brief, this study indicates that IPC confers neuroprotection against TCI by TNF-α signaling through TNF-R2 and suggests that the enhancement of TNF-R2 expression by IPC may be a legitimate strategy for a therapeutic intervention of TCI. [ABSTRACT FROM AUTHOR]

Formation of hybrid cells by "accidental cell fusion" of normal and neoplastic breast epithelial cells with local tissue‐associated mesenchymal stroma/stem‐like cells (MSC) in an inflammatory microenvironment can generate new cancer cell populations whereby molecular signaling mechanisms of this process remain unclear. Fusions of lentiviral enhanced green fluorescent protein‐labeled MSC with mcherry‐labeled breast epithelial cells were quantified and effects of tumor necrosis factor alpha (TNF‐α) and receptor downstream signaling were investigated. Cocultures of MSC with normal human mammary epithelial cells, with neoplastic MCF10A, or with MDA‐MB‐231 or MCF7 breast cancer cells demonstrated hybrid cell formation between 0.1% and about 2% of the populations within 72 hours, whereby the fusion process occurred in less than 5 minutes. Addition of the pro‐inflammatory cytokine TNF‐α significantly enhanced MCF10A‐MSC cell fusion. Small‐interfering RNA (siRNA) knockdown experiments revealed an involvement of tumor necrosis factor (TNF) receptor‐1 and ‐2 in this process. This was also substantiated by siRNA knockdown of tumor necrosis factor receptor type 1‐associated death domain which abolished TNF‐α‐stimulated fusion. While TNF receptor signaling can be relayed via the Mitogen‐activated protein kinase 8 (MAPK8), NF‐κB or cell death pathways, examination of further downstream signaling exhibited little if any effects of MAPK8 or RelA (p65) on TNF‐α‐mediated cell fusion, respectively. These data suggested that cell fusion between MSC and MCF10A breast epithelial cells can be stimulated by TNF‐α involving TNF receptor‐activated cell death pathways or additional NF‐κB signaling. Stem Cells2018;36:977–989 [ABSTRACT FROM AUTHOR]

Objective: To compare the analgesic effects and possible mechanisms of different doses of thalidomide on chronic sciatic nerve constriction (CCI) in rats. Methods: Fifty rats were randomly divided into group S. group C. group L group M. group H and group S were considered as the sham operation group. CCI model were established in the other four groups. After operation, group L. group M and group H were treated by 20 nig kg. 50 nig kg and 100 nigkg thalidomide, respectively. The mechanical pain threshold and thermal pain threshold of each group were measured and compared at 1, 2. 3 and 4 weeks after operation. Western blot and real-tune fluorescent quantitative PCR (Q-PCR) were used to analyze the niRNA and protein expressions of tumor necrosis factor receptor (TNFR). The correlation oftliaUdouiide concentration with the relative expression of TNFRmRMA was analyzed. Results: There was no significant difference in the mechanical pain threshold and thermal pain threshold in group S before and after operation (P 0.05), and the pain threshold in the other four groups was significantly lower than that before operation (P<0.05). The mechanical pain threshold was significantly increased in group C at die fourth week (P<0.05). but there was no significant difference in mechanical pain threshold and thermal pain threshold at other time points after operation (P<0.05). The mechanical pain threshold and thermal pain threshold of group L. group M and group H increased over time, and the differences were statistically significant (P<0.05). After operation, the mechanical pain threshold and thermal pain threshold in group C were significantly lower than those in group S (P=0.000). and also were significantly lower than those in group L (P=0.000). However, in the comparison of mechanical pain threshold and thermal pain threshold among different groups, group H was higher than group M (P=0.000). and group M was higher than group L (P=0.000). Compared with group C, the relative content of TNFR1 mRNA and protein in group L. group M and group H decreased significantly (P<0.05). especially in H group and M group. Pearson correlation analysis showed mat the increase of thalidomide concentration was negatively correlated with TNFR1 expression (r=-0.497, P=0.036). Conclusion: Thalidomide may exert an analgesic effect on the chronic constriction of the sciatic nerve in rats by affecting the expression of TNFR. and its analgesic effect may be enhanced with the increase of the dosage. Thalidomide is hoped to be one of the auxiliary analgesic drugs for neuropathic pain. [ABSTRACT FROM AUTHOR]

Increasing evidence suggests that cytokines and chemokines play crucial roles in chronic itch. In the present study, we evaluated the roles of tumor necrosis factor-alpha (TNF-α) and its receptors TNF receptor subtype-1 (TNFR1) and TNFR2 in acute and chronic itch in mice. Compared to wild-type (WT) mice, TNFR1-knockout (TNFR1-KO) and TNFR1/R2 double-KO (DKO), but not TNFR2-KO mice, exhibited reduced acute itch induced by compound 48/80 and chloroquine (CQ). Application of the TNF-synthesis inhibitor thalidomide and the TNF-α antagonist etanercept dose-dependently suppressed acute itch. Intradermal injection of TNF-α was not sufficient to evoke scratching, but potentiated itch induced by compound 48/80, but not CQ. In addition, compound 48/80 induced TNF-α mRNA expression in the skin, while CQ induced its expression in the dorsal root ganglia (DRG) and spinal cord. Furthermore, chronic itch induced by dry skin was reduced by administration of thalidomide and etanercept and in TNFR1/R2 DKO mice. Dry skin induced TNF-α expression in the skin, DRG, and spinal cord and TNFR1 expression only in the spinal cord. Thus, our findings suggest that TNF-α/TNFR1 signaling is required for the full expression of acute and chronic itch via peripheral and central mechanisms, and targeting TNFR1 may be beneficial for chronic itch treatment. [ABSTRACT FROM AUTHOR]