Combination Of Tumor Necrosis Factor Research Articles

Impact of treatment of rheumatoid arthritis on periodontal disease: A review.

Numerous studies support a bidirectional association between rheumatoid arthritis (RA), a chronic autoimmune degenerative inflammatory joint disease, and periodontitis, a chronic inflammatory disease caused by the immune reaction to bacteria organized in biofilms. RA and periodontitis are both multifactorial chronic inflammatory diseases that share common modifiable and non-modifiable risk factors. There is no cure for RA; treatment is based on lifestyle modifications and a variety of medications: nonsteroidal anti-inflammatory drugs (NSAID), glucocorticoids, and disease-modifying antirheumatic drugs (DMARDs, e.g., conventional synthetic DMARDs [csDMARDs]; biological DMARDs [bDMARD] and targeted synthetic DMARDs). There are molecular pathways of inflammation that are common to both RA and periodontitis. Thus, there is a potential effect of RA treatments on periodontitis. This systematic review aims to assess the impact of antirheumatic agents on periodontal conditions of patients suffering from both RA and periodontitis. PubMed/MEDLINE, Cochrane Library, and Embase online databases were systematically explored, and a manual search was performed to identify relevant studies published until January 2023. This review is registered in the PROSPERO database (CRD42023409006). A total of 2827 articles were identified, and 35 fulfilled the inclusion criteria. The included studies generally show a consensus that, at normal dosage, NSAID and corticosteroids have negligible impact on periodontium. Similarly, csDMARD alone or in combination with other csDMARD demonstrated no adverse effect on periodontium. Monotherapy with bDMARD had a positive effect on periodontal pocket depths and gingival inflammation in the longitudinal studies up to 6 months but showed negligible effect on the periodontium in interventional studies with a longer follow-up (9 months and 15.1 months). However, the combination of tumor necrosis factor (TNF)-α inhibitors+methotrexate (MTX) was associated with a rise in gingival inflammation. Due to the considerable heterogeneity of the study designs, a meta-analysis could not reasonably be performed. Within the limitations of the available studies, there is evidence to suggest that bDMARD monotherapy may improve the periodontal condition of RA patients with periodontal disease to a certain extent; the concomitant medication of TNF inhibitor+MTX could worsen gingival inflammation. More data are required to understand the impact of RA therapies on periodontal health.

Multifaceted Analysis of IL-23A- and/or EBI3-Including Cytokines Produced by Psoriatic Keratinocytes.

Interleukin (IL) 23 (p19/p40) plays a critical role in the pathogenesis of psoriasis and is upregulated in psoriasis skin lesions. In clinical practice, anti-IL-23Ap19 antibodies are highly effective against psoriasis. IL-39 (p19/ Epstein-Barr virus-induced (EBI) 3), a newly discovered cytokine in 2015, shares the p19 subunit with IL-23. Anti-IL-23Ap19 antibodies may bind to IL-39; also, the cytokine may contribute to the pathogenesis of psoriasis. To investigate IL23Ap19- and/or EBI3-including cytokines in psoriatic keratinocytes, we analyzed IL-23Ap19 and EBI3 expressions in psoriasis skin lesions, using immunohistochemistry and normal human epidermal keratinocytes (NHEKs) stimulated with inflammatory cytokines, using quantitative real-time polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA), and liquid chromatography-electrospray tandem mass spectrometry (LC-Ms/Ms). Immunohistochemical analysis showed that IL-23Ap19 and EBI3 expressions were upregulated in the psoriasis skin lesions. In vitro, these expressions were synergistically induced by the triple combination of tumor necrosis factor (TNF)-α, IL-17A, and interferon (IFN)-γ, and suppressed by dexamethasone, vitamin D3, and acitretin. In ELISA and LC-Ms/Ms analyses, keratinocyte-derived IL-23Ap19 and EBI3, but not heterodimeric forms, were detected with humanized anti-IL-23Ap19 monoclonal antibodies, tildrakizumab, and anti-EBI3 antibodies, respectively. Psoriatic keratinocytes may express IL-23Ap19 and EBI3 proteins in a monomer or homopolymer, such as homodimer or homotrimer.

The Immunomodulatory Potential of Copper and Silver Based Self-Assembled Metal Organic Biohybrids Nanomaterials in Cancer Theranostics.

Copper high aspect ratio structures (CuHARS) and silver cystine nanoparticles (AgCysNPs) are two unique micro/nano particles under study here that show extensive anti-cancer effects on a glioma tumor cell line. These micro/nano particles have shown potent toxicity in the presence of inflammatory stimulus (combination of tumor necrosis factor, [TNF] and lipo-polysaccharide, LPS). CuHARS with a concentration of 20 μg/ml uniquely increased the catalytic generation of nitric oxide (NO), an important contributor in the immune system. This NO was generated in a cell culture tumor microenvironment (TME) in the presence of 25 µM S-nitrosothiol (cysteine-NO) and the inflammatory stimulus. CuHARS increased the NO production by 68.75% when compared to untreated glioma cells with CysNO and inflammatory stimulus. The production of NO was significantly higher under similar circumstances in the case of normal primary structural cells like brain microvascular endothelial cells (BMVECs). The production of NO by BMVECs went up by 181.25% compared to glioma cells. This significant increase in the NO concentration could have added up to tumorigenesis but the anti-cancer effect of CuHARS was prominent enough to lower down the viability of glioma cells by approximately 20% and increased the metabolism of structural cells, BMVECs by approximately 200%. The immunomodulatory effect of NO in the TME under these circumstances in the presence of the novel micro/nano material, CuHARS has risen up compared to the effect of inflammatory stimulus alone. The potency and specific nature of these materials toward tumor cells may make them suitable candidates for cancer treatment. Successive treatment of CuHARS to glioma cells also proved to be an effective approach considering the decrease in the total count of cells by 11.84 fold in case of three successive treatments compared to a single dose which only decreased the cell count by 2.45 fold showing the dose-dependent increasing toxicity toward glioma cells. AgCysNPs are another potent nanomaterial which also proved its significant toxic nature toward tumor cell lines as demonstrated here, but their immunomodulatory response is still unclear and needs to be explored further.

Triple Oral Therapy Versus Antitumor Necrosis Factor Plus Methotrexate (MTX) in Patients with Rheumatoid Arthritis and Inadequate Response to MTX: A Systematic Literature Review

For patients with rheumatoid arthritis (RA) who have an inadequate response to methotrexate (MTX), the relative effectiveness of the combination of conventional disease-modifying antirheumatic drugs (DMARD) compared with the combination of tumor necrosis factor (TNF) inhibitors and MTX, as second-line therapy, is uncertain. The aim of this study was to compare the efficacy and tolerance of triple oral DMARD therapy versus anti-TNF agents associated with MTX in patients with RA after MTX failure. We performed a systematic search of the literature up to November 2015 in MEDLINE, Embase, the Cochrane library, and abstracts from the American College of Rheumatology (ACR) and the European League Against Rheumatism (EULAR) meetings from 2006 to 2015. Articles were included if they were of randomized controlled trials of patients receiving triple oral combination therapy (TT; MTX + sulfasalazine + hydroxychloroquine) compared with anti-TNF agents plus MTX. Treatment effects were examined by disease activity [Disease Activity Score in 28 joints (DAS28)], ACR and EULAR response criteria, structural damage by the modified total Sharp score, and functional disability by the Health Assessment Questionnaire (HAQ). Our search identified 263 articles; only 5 fulfilled the selection criteria. Analysis of ACR and EULAR response criteria, DAS28, and modified Sharp scores favored anti-TNF agents combined with MTX. Functional disability (HAQ) and rates of adverse events did not differ between treatments. In patients with RA in whom MTX has failed, the addition of a TNF antagonist to MTX may be a valid option, with better clinical outcomes and better radiographic results in the presence of poor prognostic factors. In the absence of poor prognostic factors and/or with contraindications to biologic agents, TT retains its place in the therapeutic strategy for RA in a currently restricted economic context.

Abstract B55: Chemoprevention of pancreatic cancer by targeting Kras mutations for apoptosis

Abstract Pancreatic cancer is often diagnosed at an advanced stage and has a poor prognosis. The 5-year survival rate is less than 5% with most patients dying within one year. Therefore, more effective approaches to treat advanced pancreatic cancer and chemopreventive strategies are in urgent need. Recent reports of early dissemination in pancreatic cancer stress the need for early intervention. Activating mutations of the Kras oncogene are one of the most common genetic abnormalities in pancreatic cancer, present in ~90%–95% of cases. In addition to its role in cancer progression, mutations of the Kras gene are one of the earliest genetic abnormalities observed in pancreatic cancer, supporting its role as an initiating event for pancreatic cancer formation. Therefore, mutant Kras gene or its gene product represents an obvious target for the prevention of pancreatic cancer. Unfortunately, direct inhibition of Kras or its downstream effectors has been largely ineffective in treating pancreatic cancer in clinical trials. We recently proposed a new cancer chemoprevention approach SITEP (short-term intermittent therapy to eliminate premalignancy) to target the “drivers” of tumorigenesis to overcome the challenges in cancer chemoprevention. We demonstrated that a two-drug combination of tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) and a small molecule mimic of the mitochondrial protein Smac /DIABLO (Smac), through a synthetic lethal interaction, specifically targets mutant Kras expressing premalignant cells for apoptosis without harming normal cells. We further showed that short-term, intermittent in vivo treatment with TRAIL and Smac mimic induced apoptosis in tumor cells and reduced tumor burden in a murine model of Kras-induced lung cancer. In this study, we examined whether targeting Kras mutant cells for apoptosis in pancreatic precursor lesions (e.g., PanINs) by TRAIL and Smac mimic would be an effective approach for pancreatic cancer chemoprevention. We showed that constitutive expression of mutant Kras led to the activation of downstream signaling events in the immortalized human pancreatic normal epithelial (HPNE) cells, including phosphorylation of ERK and AKT. Control cells are resistant to either TRAIL, or Smac mimic, or TRAIL and Smac mimic combination. The expression of Kras mutant sensitized HPNE cells to the combination of TRAIL and Smac mimic. Short term treatment of Pdx1-Cre, LSL-KrasG12D mice showed that TRAIL and Smac mimic induced significant apoptosis in abnormal pancreatic ductal cells and reduced number of PanINs. Similar short-term and intermittent treatment studies are ongoing in Pdx1-Cre, LSL-KrasG12D, LSL-p53 mouse model. We conclude that targeting oncogenic Kras signaling pathways in precursor lesions by TRAIL and Smac mimic may prove useful in developing chemoprevention against pancreatic cancer. Citation Format: Oksana Zagorodna, Huamin Wang, Xiangwei Wu. Chemoprevention of pancreatic cancer by targeting Kras mutations for apoptosis. [abstract]. In: Proceedings of the Thirteenth Annual AACR International Conference on Frontiers in Cancer Prevention Research; 2014 Sep 27-Oct 1; New Orleans, LA. Philadelphia (PA): AACR; Can Prev Res 2015;8(10 Suppl): Abstract nr B55.

Abstract B82: Chemoprevention of pancreatic cancer by targeting Kras mutations for apoptosis

Abstract Pancreatic cancer is often diagnosed at an advanced stage and has a poor prognosis. The 5-year survival rate is less than 5% with most patients dying within one year. Therefore, more effective approaches to treat advanced pancreatic cancer and chemopreventive strategies are in urgent need. Recent reports of early dissemination in pancreatic cancer stress the need for early intervention. Activating mutations of the Kras oncogene are one of the most common genetic abnormalities in pancreatic cancer, present in ~90%–95% of cases. In addition to its role in cancer progression, mutations of the Kras gene are one of the earliest genetic abnormalities observed in pancreatic cancer, supporting its role as an initiating event for pancreatic cancer formation. Therefore, mutant Kras gene or its gene product represents an obvious target for the prevention of pancreatic cancer. Unfortunately, direct inhibition of Kras or its downstream effectors has been largely ineffective in treating pancreatic cancer in clinical trials. We recently proposed a new cancer chemoprevention approach SITEP (short-term intermittent therapy to eliminate premalignancy) to target the “drivers” of tumorigenesis to overcome the challenges in cancer chemoprevention. We demonstrated that a two-drug combination of tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) and a small molecule mimic of the mitochondrial protein Smac /DIABLO (Smac), through a synthetic lethal interaction, specifically targets mutant Kras expressing premalignant cells for apoptosis without harming normal cells. We further showed that short-term, intermittent in vivo treatment with TRAIL and Smac mimic induced apoptosis in tumor cells and reduced tumor burden in a murine model of Kras-induced lung cancer. In this study, we examined whether targeting Kras mutant cells for apoptosis in pancreatic precursor lesions (e.g., PanINs) by TRAIL and Smac mimic would be an effective approach for pancreatic cancer chemoprevention. We showed that constitutive expression of mutant Kras led to the activation of downstream signaling events in the immortalized human pancreatic normal epithelial (HPNE) cells, including phosphorylation of ERK and AKT. Control cells are resistant to either TRAIL, or Smac mimic, or TRAIL and Smac mimic combination. The expression of Kras mutant sensitized HPNE cells to the combination of TRAIL and Smac mimic. Short term treatment of Pdx1-Cre, LSL-KrasG12D mice showed that TRAIL and Smac mimic induced significant apoptosis in abnormal pancreatic ductal cells and reduced number of PanINs. Similar short-term and intermittent treatment studies are ongoing in Pdx1-Cre, LSL-KrasG12D, LSL-p53 mouse model. We conclude that targeting oncogenic Kras signaling pathways in precursor lesions by TRAIL and Smac mimic may prove useful in developing chemoprevention against pancreatic cancer. Citation Format: Oksana Zagorodna, Huamin Wang, Xiangwei Wu. Chemoprevention of pancreatic cancer by targeting Kras mutations for apoptosis. [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Innovations in Research and Treatment; May 18-21, 2014; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2015;75(13 Suppl):Abstract nr B82.

AB0099 Neutrophilic Chemoattractants as Well as Monocyte Chemoattractant Protein-1 (MCP-1) and Interleukin (IL-) 6 Are Induced in Synoviocytes Stimulated with IL-17: Implications for the Pathogenesis of Rheumatoid Arthritis

BackgroundWe previously reported that helper T (Th) cells that produce interleukin (IL-) 17 by stimulation with IL-23 promote the differentiation of osteoclasts that resorb the bone matrix1. This effect was...

Combination of Tumor Necrosis Factor α and Interleukin‐6 Induces Mouse Osteoclast‐like Cells With Bone Resorption Activity Both In Vitro and In Vivo

To clarify the function of osteoclast-like multinuclear cells differentiated from bone marrow-derived macrophages (BMMs) by a combination of tumor necrosis factor α (TNFα) and interleukin-6 (IL-6), and to investigate the molecular mechanisms underlying the differentiation. BMMs were stimulated by TNFα and/or IL-6. The cells were then compared with conventional osteoclasts differentiated in vitro by RANKL. An in vitro pit formation assay on dentine slices and an in vivo resorption assay of calvarial bones were performed. We also evaluated the activities and expression levels of NF-κB, c-Fos, and NF-ATc1, which are essential to the differentiation of conventional osteoclasts. Small interfering RNA was used to knock down c-Fos. The effects of genetic ablation of STAT-3 and pharmacologic inhibitors of NF-AT, JAK, and ERK were also studied. Osteoclast-like cell differentiation depended on TNFα and IL-6 and was not inhibited by osteoprotegerin. These differentiated cells were associated with both in vitro and in vivo bone resorption activity. TNFα and IL-6 had a synergistic effect on the activity and expression of c-Fos. Knockdown of c-Fos inhibited the expression of NF-ATc1 and the differentiation of osteoclast-like cells. All of these inhibitors blocked differentiation of the cells in vitro, but surprisingly, the conditional knockout of STAT-3 did not. Tofacitinib also inhibited the bone destruction caused by TNFα and IL-6 in vivo. Our results demonstrate that a combination of the inflammatory cytokines TNFα and IL-6 can induce osteoclast-like cells that have in vitro and in vivo bone-resorptive activity.

Three-Dimensional Model of Mouse Epidermis for Experimental Studies of Psoriasis

Three-dimensional models of skin and epidermis imitate the structure of real tissues and provide accurate information about certain skin conditions, such as psoriasis. A three-dimensional model of mouse epidermis was generated from the epidermal keratinocytes of newborn mice and treated with cytokines. The aim of this study was to evaluate this model as an experimental model of psoriasis and to assess the changes occurring in its structure and gene expression after the exposure to proinflammatory cytokines. Treatment of the three-dimensional model with either interleukin 17 or a combination of tumor necrosis factor and interferon γ was shown to produce morphological changes, which were similar to acanthosis in psoriatic skin. The observed changes in gene expression of metalloproteinases and certain psoriasis biomarkers, such as mki67, krt16 and fosl1, were similar to the changes in patients' skin. Notably, changes caused by interleukin 17 were less evident than those caused by the combination of interferon γ and tumor necrosis factor. On the contrary, HaCaT cells exhibited no significant changes in the expression of fosl1 and had decreased levels of mki67 after being treated with a combination of TNF and IFNG. Moreover, treatment with IL17 had no significant effect on krt16 and mki67 expression and even reduced the fosl1 levels. The findings suggest that artificially generated three-dimensional models of murine skin can be used to study psoriasis.

Abstract 1054: Targeting EGFR signaling by synthetic lethality for chemoprevention of lung cancer.

Abstract Background: Despite continuing improvement in treatment strategy, lung cancer remains the leading cause of cancer death in the US with a low 5-year survival rate. The epidermal growth factor receptor (EGFR) plays an important role in carcinogenesis and tumour progression through activation mechanisms including overexpression, mutation, and autocrine ligand production. In the United States, activating EGFR mutations including L858R are estimated to occur in 15% of patients with primary lung adenocarcinomas. We recently proposed a new cancer chemoprevention approach SITEP (short-term intermittent therapy to eliminate premalignancy) to target the “drivers” of tumorigenesis to overcome the challenges in cancer chemoprevention. In a scientific proof-of-concept study, we demonstrated that a two-drug combination of tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) and a small molecule mimic of the mitochondrial protein Smac /DIABLO (Smac), through a synthetic lethal interaction, specifically targets the activation of KRAS signaling for lung cancer chemoprevention (Huang et al., CaPR 2011). Since activation of EGFR leads to the activation of KRAS signaling, we hypothesize that TRAIL and Smac mimic will specifically target mutant EGFR for apoptosis through synthetic lethality and can be used for lung cancer chemoprevention. Thus the aim of this study was to elucidate the mechanism and evaluate the efficacy of TRAIL and Smac mimic on the induction of apoptosis in EGFR (L585R) mutant lung cancer cells in vitro. Methods: To explore the mechanism of EGFR-L858R-mediated sensitization to TRAIL and Smac in vitro, a EGFR-L858R expressing plasmid was transfected into normal immortalized lung epithelial cells. Activation of downstream signaling molecules MAPK and the AKT was assessed. Modulation of TRAIL receptor expression was analyzed. The effect on TRAIL and Smac-mediated apoptosis was measured. Results: As expected, the overexpression of EGFR-L858 receptor leads to an activation of ERK and AKT by phosphorylation, an activation of c-Myc and a repression of c-FLIP. More importantly, expression of EGFR-L858R sensitized normal epithelial cells to TRAIL and Smac mimic-induced apoptosis. Various inhibitors targeting the MAPK and PI3K-AKT pathways are used to confirm this effects and the contribution of these pathways on apoptosis. Conclusion: Our results showed that TRAIL and Smac mimic selectively induced apoptosis in EGFR-L858R expressing lung epithelial cells in vitro. Further studies are required to elucidate detailed mechanism, and to test the efficiency of this selective therapeutic approach in the chemoprevention of lung cancer in animal models. Citation Format: Yannis Hara, Shaoyi Huang, Xiangwei Wu. Targeting EGFR signaling by synthetic lethality for chemoprevention of lung cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1054. doi:10.1158/1538-7445.AM2013-1054

Abstract 1076: Chemoprevention of pancreatic cancer by targeting Kras mutations for apoptosis.

Abstract Pancreatic cancer is often diagnosed at an advanced stage and has a poor prognosis. The overall survival rate is less than 4% at 5 years with most patients dying within one year. Therefore, more effective approaches to treat advanced pancreatic cancer and chemopreventive strategies are in urgent need. Recent reports of early dissemination in pancreatic cancer stress the need for early intervention. Activating mutations of the KRAS oncogene are possibly the single most common genetic abnormality in pancreatic cancer, present in ∼90%-95% of cases. In addition to its role in cancer progression, mutations of the KRAS gene are one of the earliest genetic abnormalities observed in pancreatic cancer, supporting its role as an initiating event for pancreatic cancer formation. Therefore, mutant KRAS gene or its gene product represents an obvious target for the prevention of pancreatic cancer. Unfortunately, direct inhibition of KRAS or its downstream effectors has been largely ineffective in treating pancreatic cancer in clinical trials. The ideas are to find an innovative approach to target KRAS. We recently proposed a new cancer chemoprevention approach SITEP (short-term intermittent therapy to eliminate premalignancy) to target the “drivers” of tumorigenesis to overcome the challenges in cancer chemoprevention. We demonstrated that a two-drug combination of tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) and a small molecule mimic of the mitochondrial protein Smac /DIABLO (Smac), through a synthetic lethal interaction, specifically targets mutant KRAS expressing premalignant cells for apoptosis without harming normal cells. We further showed that short-term, intermittent in vivo treatment with TRAIL and Smac mimic induced apoptosis in tumor cells and reduced tumor burden in a murine model of Kras-induced lung cancer. In this study, we examined whether targeting KRAS mutant cells for apoptosis in pancreatic precursor lesions (e.g., PanINs) by TRAIL and Smac mimic would be an effective approach for pancreatic cancer chemoprevention. We showed that constitutive expression of mutant KRAS led to the activation of downstream signaling events in the immortalized human pancreatic normal epithelial (HPNE) cells, including phosphorylation of ERK and AKT. TRAIL and Smac mimic treatment showed that control cells are resistant to either TRAIL, or Smac mimic, or TRAIL and Smac mimic combination, and expression of KRAS mutant sensitized cells to the combination of TRAIL and Smac mimic. Short term treatment of Pdx1-Kras mice showed that TRAIL and Smac mimic induced significant apoptosis in abnormal pancreatic ductal cells and reduced number of PanINs. We conclude that targeting oncogenic KRAS signaling pathways in precursor lesions by TRAIL and Smac mimic may prove useful in developing chemoprevention against pancreatic cancer. Citation Format: Oksana Zagorodna, Shaoyi Huang, Huamin Wang, Xiangwei Wu. Chemoprevention of pancreatic cancer by targeting Kras mutations for apoptosis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1076. doi:10.1158/1538-7445.AM2013-1076

CXCR6 and CXCL16 in liver disease

BackgroundChemokines are small molecules that act through G-protein coupled receptors to mediate primarily lymphocyte migration. CXCL16, which interacts with only one receptor (CXCR6), can mediate lymphocyte recruitment and has been implicated in various disease conditions. Steatohepatitis, caused by metabolic syndrome or alcohol misuse, is the commonest cause of liver disease in the UK. We investigated the role of CXCL16 and CXCR6 in the development of steatohepatitis. MethodsExpression of CXCL16 in whole liver and isolated cells was investigated with real-time PCR and immunohistochemistry. Serum and supernatant concentrations of soluble CXCL16 were measured with ELISA. Expression of CXCR6 on lymphocytes was investigated with flow cytometry. Lymphocyte adhesion was assessed with freshly isolated lymphocytes from liver or peripheral blood flowed over confluent layer of isolated human hepatic sinusoidal endothelium (HSEC). FindingsWhole liver expression of CXCL16 was increased relative to normal liver in fatty liver disease with increasing expression seen with increasing steatohepatitis and fibrosis. Immunohistochemistry showed CXCL16 expressed throughout regenerative nodules in both alcoholic and non-alcoholic liver disease. Isolated HSEC, biliary epithelial cells, and hepatoma cell lines increased expression of CXCL16 and released soluble CXCR6 in response to pro-inflammatory cytokines, particularly the combination of tumour necrosis factor α and interferon γ. Peripheral blood lymphocyte CXCR6 expression was confined to CD4 cells; however in the liver CD8+ cells and CD56+ cells more commonly expressed CXCR6. Inhibition of CXCR6 or CXCL16 inhibited transmigration of lymphocytes across HSEC. InterpretationCXCL16 is expressed in diseased liver where it has a role in the transmigration of lymphocytes across endothelium. This may represent a new therapeutic target in liver disease. FundingUK Medical Research Council.

Plant polyphenols differentially modulate inflammatory responses of human keratinocytes by interfering with activation of transcription factors NFκB and AhR and EGFR–ERK pathway

Molecular mechanisms underlying modulation of inflammatory responses in primary human keratinocytes by plant polyphenols (PPs), namely the glycosylated phenylpropanoid verbascoside, the stilbenoid resveratrol and its glycoside polydatin, and the flavonoid quercetin and its glycoside rutin were evaluated. As non-lethal stimuli, the prototypic ligand for epidermal growth factor receptor (EGFR) transforming growth factor alpha (TGFalpha), the combination of tumor necrosis factor (TNFalpha) and interferon (IFNgamma) (T/I), UVA+UVB irradiation, and bacterial lipopolysaccharide (LPS) were used. We demonstrated differential modulation of inflammatory responses in keratinocytes at signal transduction, gene transcription, and protein synthesis levels as a function of PP chemical structure, the pro-inflammatory trigger used, and PP interaction with intracellular detoxifying systems.The PPs remarkably inhibited constitutive, LPS- and T/I-induced but not TGFalpha-induced ERK phosphorylation. They also suppressed NFkappaB activation by LPS and T/I. Verbascoside and quercetin invariably impaired EGFR phosphorylation and UV-associated aryl hydrocarbon receptor (AhR)-mediated signaling, while rutin, polydatin and resveratrol did not affect EGFR phosphorylation and further activated AhR machinery in UV-exposed keratinocytes. In general, PPs down-regulated gene expression of pro-inflammatory cytokines/enzymes, except significant up-regulation of IL-8 observed under stimulation with TGFalpha. Both spontaneous and T/I-induced release of IL-8 and IP-10 was suppressed, although 50μM resveratrol and polydatin up-regulated IL-8. At this concentration, resveratrol activated both gene expression and de novo synthesis of IL-8 and AhR-mediated mechanisms were involved. We conclude that PPs differentially modulate the inflammatory response of human keratinocytes through distinct signal transduction pathways, including AhR and EGFR.

Effect of Th1/Th2 Cytokine Pretreatment on RSV-Induced Gene Expression in Airway Epithelial Cells

Background: Respiratory syncytial virus (RSV) infection in infants with Th2 predisposition is thought to increase the risk of allergic sensitization, recurrent wheezing, and bronchial asthma during childhood. We attempted to clarify the molecular mechanisms by which Th1/Th2 predisposition in the host alters RSV infection and facilitates airway inflammation. Methods: A549 human airway epithelial cells were inoculated with live or UV-treated RSV after pretreatment with either a combination of tumor necrosis factor (TNF)-α and interferon-γ (Th1-primed) or a combination of TNF-α and interleukin-4 (Th2-primed) for 48 h. The gene and protein expression profiles of RSV-infected A549 cells were examined. Results: GeneChip analysis indicated that, at 96 h after inoculation with RSV, the expression of 62 genes was specifically enhanced (more than 2-fold by normalized data) in Th2-primed cells compared to that in unprimed or Th1-primed cells. An increase in mRNA and protein levels of monocyte chemoattractant protein (MCP)-1/CCL2 among those 62 genes was confirmed by real-time PCR and cytometric bead assay, respectively. RSV replication was markedly diminished in Th1-primed airway epithelial cells but not in Th2-primed cells, which was presumably caused at least in part by the early induction of antiviral genes. Conclusions: These results suggest that Th1/Th2 predisposition in the host prior to RSV infection critically regulates inflammatory reactions in the airways through alteration of gene expression, and that MCP-1/CCL2 plays an important role in the pathogenesis of severe RSV infection and the subsequent development of asthma in Th2-predisposed hosts.

Cytokine-mediated Regulation of CX3CL1 in Osteoblasts from Patients with Rheumatoid Arthritis

IntroductionCX3CL1 (fractalkine), a membrane-bound chemokine that induces both the adhesion and migration of leukocytes, is involved in the recruitment of cells to tissues undergoing inflammatory responses. To explore the regulation of CX3CL1 in inflammatory bone diseases, we examined CX3CL1 expression in osteoblasts.MethodsHuman osteoblasts isolated from the femora of rheumatoid arthritis patients were incubated in the presence or absence of various inflammatory stimuli. Culture supernatants were collected, and soluble CX3CL1 levels were determined with an enzyme-linked immunosorbent assay (ELISA). The expression of CX3CL1 mRNA transcripts in osteoblasts was examined using the quantitative TaqMan real-time polymerase chain reaction.ResultsThe combination of tumor necrosis factor (TNF)-α and interferon (IFN)-γ induced dramatic increases in levels of both soluble CX3CL1 protein and mRNA transcripts. CX3CL1 expression in osteoblasts was decreased by the addition of interleukin(IL)-4 or IL-17 but was increased when stimulation by IFN-γ and IL-17 was supplemented with IL-1β In addition, expression was decreased when TNF-α was added.ConclusionsMultiple cytokines, including IL-17, are able to either increase or decrease the expression of CX3CL1 by human osteoblasts.

The Anti‐Inflammatory Role of Heme Oxygenase‐1 in Lipopolysaccharide and Cytokine‐Stimulated Inducible Nitric Oxide Synthase and Nitric Oxide Production in Human Periodontal Ligament Cells

Although heme oxygenase-1 (HO-1) is involved in anti-inflammation, the mechanisms of its activity in regulating periodontal inflammation are largely unclear. Therefore, the aim of this study is to investigate the anti-inflammatory properties of HO-1 in lipopolysaccharide (LPS)- and proinflammatory cytokine-stimulated inducible nitric oxide synthase (iNOS) expression and nitric oxide (NO) production in human periodontal ligament (PDL) cells. PDL cells were treated with LPS plus a combination of tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta in serum-free media for 1 day. The production of NO was evaluated using a Griess reagent kit. The expression of iNOS and HO-1 proteins and mRNAs was evaluated using Western blotting and reverse transcriptase-polymerase chain reaction, respectively. Proinflammatory cytokines and LPS triggered iNOS and HO-1 expression and NO production in PDL cells. HO-1 inhibitor and HO-1 small interfering RNA (siRNA) attenuated the LPS- and cytokine-stimulated NO release and iNOS and HO-1 expression. Specific inhibitors of p38, extracellular signal-regulated kinase (ERK), and c-Jun N-terminal kinase (JNK) mitogen-activated protein kinases phosphatidylinositol 3-kinase (PI3K), nuclear factor-kappa B (NF-kappaB), and protein kinase C delta (PKC-delta) greatly reduced the levels of iNOS and HO-1 expression induced by LPS plus cytokines. Collectively, these data suggested that HO-1 inhibition blocked LPS- and proinflammatory cytokine-stimulated iNOS expression and NO production in PDL cells via a mechanism that involves p38, ERK, PI3K, NF-kappaB, and PKC-delta. Thus, the regulation of HO-1 activity may be a therapeutic strategy for periodontal disease.

Disease remission state in patients treated with the combination of tumor necrosis factor blockade and methotrexate or with disease‐modifying antirheumatic drugs: A clinical and imaging comparative study

For patients with rheumatoid arthritis (RA) in remission who are receiving disease-modifying antirheumatic drugs (DMARDs), radiographic progression correlates with imaging-detected synovitis as measured by power Doppler activity. In contrast, patients with disease in remission who are receiving the combination of tumor necrosis factor (TNF) blockade with methotrexate (MTX) (combination treatment) have reduced radiographic damage for the equivalent clinical state. We undertook this study to determine whether the difference in radiographic outcome is a result of more complete suppression of imaging-detected synovitis. One hundred patients with RA in remission (Disease Activity Score in 28 joints [DAS28] <2.6) for at least 6 months while receiving either combination treatment (n = 50) or DMARDs (n = 50) were matched for clinical variables. Ultrasound of metacarpophalangeal joints 1-5 and the wrist joints was performed. Remission according to imaging results was defined as a score of 0 for both grey scale synovitis and power Doppler activity. In patients receiving combination treatment or DMARDs (median DAS28 1.65 versus 1.78, median disease duration 120 months versus 90 months, and median duration of remission 13 months versus 18 months), the proportion with remission according to imaging results was not significantly different (10% versus 16%, respectively). The combination treatment group had more grey scale synovitis (P < 0.001) but similar power Doppler activity (48% versus 60%, respectively; P = 0.229) in any joint as compared with the DMARD group. Results were not affected by stratification for duration of disease or remission. In RA patients with disease in remission, imaging-detected synovitis persists, with power Doppler activity seen in >or=48% of the patients regardless of therapy. These results suggest that superior radiographic outcomes in patients treated with the combination of TNF blockade and MTX may not be due to complete suppression of imaging-detected synovitis.

Adding low dose tacrolimus in rheumatoid arthritis patients with an inadequate response to tumor necrosis factor inhibitor therapies

In the present study, we retrospectively evaluate the efficacy of low dose tacrolimus (TAC) as add-on therapy in refractory rheumatoid arthritis (RA) despite a combination of tumor necrosis factor (TNF) inhibitor and methotrexate (MTX) using consecutive case series of five patients with active RA (mean disease duration 2.3 years) despite MTX and TNF inhibitors for at least 3 months (mean 9.5 months) treated with low dose TAC (1.5-2 mg/day) for at least 6 months (mean 1.8 years). Clinical and radiographic efficacy was assessed according to the European league against rheumatism response criteria and the modified Sharp method, respectively. At 1 year, three patients reached to remission. The mean yearly progression of radiographic joint damage of all five patients after the onset of TAC was significantly decreased compared to that observed during anti-TNF therapy without TAC (p = 0.04). One patient temporally discontinued the treatment because of herpes zoster. In RA patients with inadequate response to MTX and a TNF inhibitor, additions of low dose TAC markedly improved clinical variables including radiographic scores without remarkable detrimental effects. It seems that TAC in combination with MTX and TNF inhibitors may be a hopeful treatment option for RA patients with inadequate response to anti-TNF therapy.

Comparative effectiveness of tumour necrosis factor α inhibitors in combination with either methotrexate or leflunomide

Objective:The objective of this study was to compare the effectiveness of a combination of tumour necrosis factor α (TNFα) inhibitors with either methotrexate or leflunomide in the treatment of patients...

Essential Role of the JAK/STAT1 Signaling Pathway in the Expression of Inducible Nitric-oxide Synthase in Intestinal Epithelial Cells and Its Regulation by Butyrate

Nitric oxide (NO) is a highly reactive free radical that modulates tumorigenesis through its ability to regulate cell proliferation, cell death, migration and angiogenesis. Although the role of NO has been well studied in inflammatory cells, much less is known about the regulation of NO production in epithelial cells. We demonstrated that in intestinal epithelial cells the expression of inducible NO synthase (iNOS), the critical enzyme in the synthesis of NO, is synergistically stimulated by bacterial lipopolysaccharide (LPS) and interferon gamma (IFNgamma) or by the combination of tumor necrosis factor (TNF) and IFNgamma at the transcriptional level. Expression of iNOS and the production of NO in response to LPS/IFNgamma were significantly increased upon induction of oncogenic K-Ras, underlying frequently elevated expression of iNOS in colon cancer. Silencing of STAT1, a major transcription factor involved in signaling by IFNgamma, or pharmacological inhibition of JAKs, kinases that phosphorylate STATs, prevented the induction of iNOS and the production of NO in response to stimulation of cells with LPS/IFNgamma or TNF/IFNgamma, underscoring the importance of the intact JAK/STAT signaling in the regulation of iNOS expression in intestinal epithelial cells. Butyrate, a histone deacetylase (HDAC) inhibitor and a dietary chemopreventive agent, decreased NO production in macrophages and in intestinal myofibroblasts, consistent with its anti-inflammatory activity. In contrast, in intestinal epithelial cells, butyrate significantly enhanced the expression of iNOS and the production of NO in response to treatment with LPS/IFNgamma. Despite the fact that, like butyrate, three structurally unrelated inhibitors of HDAC activity, trichostatin A, suberoylanilide hydroxamic acid, and apicidin, induced acetylation of H3 and H4 in epithelial cells, they failed to increase the production of NO, demonstrating that butyrate regulates NO production in epithelial cells in an HDAC-independent manner. The ability of butyrate to regulate the production of NO in a variety of cell types is likely to underlie its potent chemopreventive and anti-inflammatory activity.