Search

Psoriasis is a recrudescent chronic immune-mediated inflammatory dermatosis; the production and release of proinflammatory cytokines/chemokines such as TNF-a has been regarded as critical issues during psoriasis pathogenesis. Based on online microarray profiles, the expression of the transcription factor GATA3 was downregulated in psoriasis lesion tissues. In the present study, we searched for miRNAs that might be related to TNF-a and GATA3 to investigate an in-depth understanding of psoriasis pathogenesis. Herein, higher TNF-a and GATA3 protein levels were observed in psoriasis lesion tissues and that GATA3 overexpression significantly reverses TNF-a-induced increases within the production of IL-6 and CXCL8 in keratinocytes. TNF-a stimulation increases miR-155 expression dose-independently, and the miR-155 inhibitor significantly reverses TNF-a-induced suppression of GATA3 protein levels and increases IL-6 and CXCL8 production. miR-155 could suppress the expression of GATA3 by targeting its 3'UTR, while GATA3 could activate the transcription of IL37 by targeting its promoter region. miR-155 overexpression reduces IL37 protein and increases CXCL8 production; GATA3 overexpression might significantly attenuate the effects of miR-155 overexpression. In contrast to GATA3, miR-155 expression is significantly upregulated in psoriasis lesion tissue and is negatively correlated with GATA3 and IL37. In summary, the miR-155/GATA3/IL37 axis modulates the production of IL-6 and CXCL8 upon TNF-a stimulation to affect psoriasis development. Thus, miR-155/GATA3/IL37 may be potent targets for psoriasis treatment, which needs further in vivo and clinical investigation. [ABSTRACT FROM AUTHOR]

Bone homeostasis is preserved by the balance of maintaining between the activity of osteogenesis and osteoclastogenesis. However, investigations for the osteoclastogenesis were hampered by considerable difficulties associated with isolating and culturing osteoclast in vivo. As the alternative, stimuli‐induced osteoclasts formation from RAW264.7 cells (RAW‐OCs) have gain its importance for extensively osteoclastogenic study of bone diseases, such as rheumatoid arthritis, osteoporosis, osteolysis and periodontitis. However, considering the RAW‐OCs have not yet been well‐characterized and RAW264.7 cells are polymorphic because of a diverse phenotype of the individual cells comprising this cell linage, and different fate associated with various stimuli contributions. Thus, in present study, we provide an overview for current knowledge of the phenotype of RAW264.7 cells, as well as the current understanding of the complicated interactions between various stimuli and RAW‐OCs in the light of the recent progress. [ABSTRACT FROM AUTHOR]

ABSTRACT [ABSTRACT FROM AUTHOR]

Little is known about the immunologic maturation in the early stages of life. The aim of this study was to investigate maturation of immune system from birth to 1 yr of age and to compare immune functions between mothers and their children. Also the effect of atopy to the immune responses of children was examined. Cord blood samples (n = 228) and peripheral blood samples of children (n = 200) and their mothers (n = 208) 1 yr after birth were collected. Whole blood samples were stimulated for 24 and 48 h with Staphylococcal enterotoxin B (SEB), lipopolysaccharide (LPS) and the combination of phorbol ester and ionomycin (P/I). Production of TNF-α, IFN-γ, IL-5, IL-8 and IL-10 was determined using ELISA. Significant mother-to-child correlation was detected in cytokine-producing capacity at the age of 1 yr. TNF-α (P/I, SEB and LPS stimulation), IFN-γ (P/I and SEB), IL-5 (P/I and SEB) and IL-10 (P/I, SEB and LPS) producing capacity increased from birth to 1 yr of age. In general, stimulated cytokine responses were higher in mothers’ than in children’s blood samples, except in the case of P/I and LPS-stimulated IL-8, which were highest at birth. Maternal inhalation atopy was associated with increased cord blood IL-5 (24 and 48 h) and IL-10 (48 h) production following P/I stimulation. Also children of food atopic mothers expressed elevated cord blood IL-10 (48 h, P/I) responses and decreased IFN-γ/IL-5 ratio (24 h, P/I). In addition, the production of IFN-γ (24 and 48 h, P/I) and the IFN-γ/IL-5 ratio (24 h and 48 h, P/I) at the age of 1 yr was lower among children with food atopic mothers. In conclusion, our results suggest that both adaptive and innate immune responses increase from birth to 1 yr of age, but are still weak in comparison to adult responses. Cytokine responses of children begin to correlate with those of their mothers during the first year of life. Although only few associations were observed between atopy and cytokine-producing capacity, our results suggest that children of atopic mothers express Th2-polarized cytokine pattern. [ABSTRACT FROM AUTHOR]

Objective. The purpose of the present study was to determine the effect of body weight on blood lipid profile, insulin resistance and inflammatory biomarkers in Mexican-American children. Methods. Children (13.3±0.1 year) were recruited from a local school and assigned to one of three groups as a volunteer sample: healthy weight (HW) (≥10th and <85th BMI percentile; n = 42), at risk of overweight (RO) (≥85th and <95th; n = 25) or overweight (OW) (≥95th; n = 42). Plasma concentrations of hsCRP, sCD14, sIL-6R, sTNF-αR1, sTNF-αR2, IL-6 and TNF-α were determined by ELISA. Results. OW children had significantly greater plasma concentrations of hsCRP (P = 0.003), sCD14 (P = 0.013), sIL-6R (P = 0.010), sTNF-αR1 (P < 0.001), sTNF-αR2 (P = 0.005), insulin (P = 0.001), TC:HDL ratio (P < 0.001) and triglycerides (P < 0.001) than HW children. Also plasma concentrations of hsCRP, sIL-6R and sTNF-αR1 were significantly greater in OW compared with RO children. Conclusion. Overweight Mexican-American children had a higher concentration of inflammatory biomarkers than healthy weight children. To our knowledge, this is the first study to report that sCD14 is elevated in overweight compared with healthy weight Mexican-American children. [ABSTRACT FROM AUTHOR]

Aim: To compare the frequencies of cytokine and receptor molecule genotypes in patients with chronic periodontitis with the corresponding frequencies in a reference population and to study the relationship between periodontal disease severity and polymorphisms in the studied genes. Subjects and methods: CD14, IL-6, TNF- α, IL-10, IL-1 α, IL-1 β, and TLR-4 polymorphisms of 51 periodontitis patients were studied using polymerase chain reaction. The genotype frequencies in the periodontitis patients and a reference population ( n=178) were compared. Probing pocket depth (PD), periodontal attachment level (AL), and alveolar bone level (BL) were related to the genotypes. Results: No statistically significant differences could be found between the frequencies of the cytokine genotypes in the periodontitis patients and in the reference group. The extent of periodontal disease was higher in subjects with the T-containing genotype of CD14−260 and the GG genotype of IL-6−174 when compared with the extent in the rest of the group. Subjects carrying the composite genotype of the above two were most severely affected by periodontal disease. Conclusion: According to the present results, an evident association exists between the carriage of the T-containing genotype of CD14−260 and the GG genotype of IL-6−174 and the extent periodontal disease. [ABSTRACT FROM AUTHOR]

Aim To determine the influence of Type 1 diabetes mellitus on circulating adipocytokines in children. Methods The circulating concentrations of leptin, adiponectin, resistin and tumour necrosis factor (TNF)-α were measured in 91 children, aged 11.1 ± 2.7 years, with Type 1 diabetes mellitus (T1DM). Ninety-one healthy children were selected as control subjects. Results Body mass index-adjusted leptin concentrations were higher in the pubertal diabetic children compared with the control children. There was a significant positive correlation between leptin and daily insulin dose in the diabetic group. Circulating adiponectin concentrations were higher in the prepubertal diabetic children and were positively associated with HbA1c. Resistin concentrations were lower in the prepubertal non-diabetic subjects compared with the pubertal non-diabetic children, whose values were higher than those of the diabetic children. TNF-α concentrations were similar in non-diabetic and diabetic children. Conclusions Circulating concentrations of adipocytokines are abnormal in Type 1 diabetic children, although the direction of change differs by cytokine. Pubertal development, in addition to insulin treatment and glycaemic control, also influences the concentrations. [ABSTRACT FROM AUTHOR]

Objective: To evaluate whether or not genetic polymorphism of IL-1B, IL-1RN, and TNF-Α is an important factor in the different expression of gastroduodenal diseases after Helicobacter pylori infection. Methods: This study consisted of 1360 subjects: control, gastric cancer (GC, intestinal type, and diffuse type), benign gastric ulcer (BGU), duodenal ulcer, and first-degree gastric cancer relative (GCR). IL-1Β-511 and TNF-A-308 biallelic polymorphism were genotyped by 5′ nuclease polymerase chain reaction (PCR) assays, and PCR-restriction fragment length polymorphism (PCR-RFLP). IL-1RN penta-allelic variable number of tandem repeats was genotyped by PCR. Results: There was no difference in the genetic polymorphism of IL-1Β-511, IL-1RN and TNF-A in the patients with gastric cancer regardless of H. pylori positivity compared with control. However, the frequencies of IL-1B-511 C/T (OR: 0.5, 95% CI: 0.3–0.7) and T carrier (OR: 0.6, 95% CI: 0.4–0.8) were lower in the H. pylori-positive BGU patients. The IL-1RN 2/2 was higher (OR: 5.5, 95% CI: 1.1–28.5) in the H. pylori-positive GCR. There was no significance in the polymorphism of TNF-Α-308 regardless of H. pylori-induced gastroduodenal diseases. Conclusions: The IL-1Β-511 T-carrier polymorphism has a negative effect on the development of H. pylori-positive BGU, and high frequency of IL-1RN 2/2 was found in the H. pylori-positive relatives of GC patients, which suggest that this genetic polymorphism could play some role in the H. pylori-induced gastroduodenal diseases in Korea. [ABSTRACT FROM AUTHOR]

Abstract: Tumor necrosis factor-alpha (TNF-a) is a major mediator of inflammatory response in many diseases. It inhibits bone formation and stimulates bone resorption. To determine the molecular mechanisms involved in the regulation of gene expression of osteoblast-like cells, we analyzed the effects of TNF-a on the human osteosarcoma cell line Saos2. We used RT-PCR to examine the effects of TNF-a on bone sialoprotein (BSP), core binding factor a1 (Cbfa1), osterix, a1 (I) collagen, cyclooxygenase-2 (COX-2), interleukin-6 (IL-6), cathepsin B, cathepsin L and tissue inhibitors of metalloproteinase-1 (TIMP-1). TNF-a (10ng/ml) increased BSP, IL-6 and COX-2 mRNA levels after 3h, reaching maximal levels at 12h. Cbfa1 mRNA levels increased after 3h, but decreased by 24h. Osterix, cathepsin B, cathepsin L and TIMP-1 mRNA levels did not change after stimulation with TNF-a. On the other hand, a1 (I) collagen mRNA expression was suppressed by TNF-a at 24h. Transient transfection analyses were performed using chimeric constructs of the rat BSP gene promoter linked to a luciferase reporter gene. TNF-a (10ng/ml) had no effect on the promoter activities of BSP transfected into Saos2 cells. The results of gel mobility shift assays using radiolabeled double-stranded cAMP response element (CRE) and FGF2 response element (FRE) oligonucleotides in the proximal promoter of the rat BSP gene showed increased binding of nuclear proteins at 6h. Gel mobility shift assays with radiolabelled COX-2-CRE and COX-2-NF¿B oligonucleotides revealed an increase in the binding of nuclear proteins from TNF-a-stimulated Saos2 cells. These studies, therefore, showed that TNF-a indirectly increased BSP expression, and that it could be mediated through COX-2 and Cbfa1 expression in Saos2 osteoblast-like cells. [Copyright &y& Elsevier]

1: The present study was undertaken to investigate the anti-inflammatory effects of a synthetic compound, LCY-2-CHO, on the expression of inducible nitric oxide synthase (iNOS), COX-2, and TNF-a in murine RAW264.7 macrophages. 2: Within 1-30?µM, LCY-2-CHO concentration-dependently inhibited lipopolysaccharide (LPS)-induced nitric oxide (NO), prostaglandin E2 (PGE2), and tumor necrosis factor-a (TNF-a) formation, with IC50 values of 2.3, 1, and 0.8?µM, respectively. Accompanying inhibition of LPS-induced iNOS, cyclooxygenase-2 (COX-2), and pro-TNF-a proteins was observed. 3: Reverse transcription-polymerase chain reaction (RT-PCR) and promoter analyses indicated that iNOS expression was inhibited at the transcriptional level (IC50=2.3?µM), that inhibition of COX-2 expression only partially depended on gene transcription (IC50=7.6?µM), and that TNF-a transcription was unaffected. 4: Transcriptional assays revealed that activation of AP-1, but not NF-?B, was concomitantly blocked by LCY-2-CHO. Our results showed that LCY-2-CHO was capable of interfering with post-transcriptional regulation, altering the stability of COX-2 and TNF-a mRNAs. 5: Since the 3'-untranslated region (3' UTR) of both COX-2 and TNF-a mRNA contains a p38 mitogen-activated protein kinase (MAPK)-regulated element involved in mRNA stability, we assessed the effect of LCY-2-CHO on p38 MAPK. Our data clearly indicated an inhibition (IC50=1.7?µM) of LPS-mediated p38 MAPK activity, but not of extracellular signal-regulated kinase (ERK) or c-Jun N-terminal kinase (JNK) activity. However, kinase assays ruled out a direct inhibition of p38 MAPK action. The selective p38 MAPK inhibitor, SB203580, inhibited the promoter activities of iNOS and COX-2 rather than that of TNF-a. 6: In conclusion, LCY-2-CHO downregulates inflammatory iNOS, COX-2, and TNF-a gene expression in macrophages through interfering with p38 MAPK and AP-1 activation.British Journal of Pharmacology (2004) 141, 1037-1047. doi:10.1038/sj.bjp.0705700 [ABSTRACT FROM AUTHOR]

Pro-inflammatory cytokines play critical roles in regulating valvular interstitial cell (VIC) phenotypic changes that can cause heart valve fibrosis and calcification. Tumor necrosis factor alpha (TNF-α) is a cytokine known to influence VIC behavior and has been reported at high levels in calcified valves ex vivo. We sought to understand the specific effects of TNF-α on VIC phenotypes (eg, fibroblast, profibrotic activated myofibroblasts) and its link with heart valve disorders. We characterize human aortic valve tissue from patients with valve disorders and identify a high variability of fibrotic and calcific markers between tissues. These results motivated in vitro studies to explore the effects of TNF-α on defined VIC fibroblasts and profibrotic activated myofibroblasts, induced via FGF-2 and TGF-β1 treatment. Using 3D hydrogels to culture VICs, we measure the effect of TNF-α (0.1-10 ng/mL) on key markers of fibrosis (eg, αSMA, COL1A1) and calcification (eg, RUNX2, BMP2, and calcium deposits). We observe calcification in TNF-α-treated VIC activated myofibroblasts and identify the MAPK/ERK signaling cascade as a potential pathway for TNF-α mediated calcification. Conversely, VIC fibroblasts respond to TNF-α with decreased calcification. Treatment of VIC profibrotic activated myofibroblast populations with TNF-α leads to increased calcification. Our in vitro findings correlate with findings in diseased human valves and highlight the importance of understanding the effect of cytokines and signaling pathways on specific VIC phenotypes. Finally, we reveal MAPK/ERK as a potential pathway involved in VIC-mediated matrix calcification with TNF-α treatment, suggesting this pathway as a potential pharmaceutical target for aortic valve disease. [ABSTRACT FROM AUTHOR]