Strong inhibition of TNF-α production and inhibition of IL-8 and COX-2 mRNA expression in monocyte-derived macrophages by RWJ 67657, a p38 mitogen-activated protein kinase (MAPK) inhibitor

In inflammatory processes, the p38 mitogen-activated protein kinase (MAPK) signal transduction route regulates production and expression of cytokines and other inflammatory mediators. Tumor necrosis factor α (TNF-α) is a pivotal cytokine in rheumatoid arthritis and its production in macrophages is under control of the p38 MAPK route. Inhibition of the p38 MAPK route may inhibit production not only of TNF-α, but also of other inflammatory mediators produced by macrophages, and indirectly of inflammatory mediators by other cells induced by TNF-α stimulation. Here we investigate the effects of RWJ 67657, a p38 MAPK inhibitor, on mRNA expression and protein production of TNF-α and other inflammatory mediators, in monocyte-derived macrophages. A strong inhibition of TNF-α was seen at pharmacologically relevant concentrations of RWJ 67657, but also inhibition of mRNA expression of IL-1β, IL-8, and cyclooxygenase-2 was shown. Furthermore, it was shown that monocyte-derived macrophages have a high constitutive production of matrix metalloproteinase 9, which is not affected by p38 MAPK inhibition. The results presented here may have important implications for the treatment of rheumatoid arthritis

Expression and regulation of HIF-1alpha in macrophages under inflammatory conditions; significant reduction of VEGF by CaMKII inhibitor

<p>Abstract</p> <p>Background</p> <p>Macrophages expressing the pro-angiogenic transcription factor hypoxia-inducible factor (HIF)-1alpha have been demonstrated in rheumatoid arthritis (RA) in the synovial tissue. Aim of the present study was to investigate intracellular signal transduction regulation of pro-inflammatory HIF-1 alpha expression in macrophages to identify possible new intervention strategies. We investigated the effects of CaMKII-inhibitors amongst other kinase inhibitors, on HIF-1 alpha expression and downstream production of pro-angiogenic factors in macrophages.</p> <p>Methods</p> <p>Differentiated THP-1 cells and synovial fluid (SF) macrophages were stimulated with 1 μg/ml LPS with or without pretreatment with specific inhibitors of the ERK pathway (PD98059), the PI3K pathway (LY294002), and the CaMKII pathway (KN93 and SMP-114). mRNA and protein expression of HIF-1 alpha, VEGF, MMP-9, and IL-8 was measured in cell lysates and cell supernatants.</p> <p>Results</p> <p>HIF-1 alpha protein expression in LPS-stimulated THP-1 macrophages could be blocked by ERK- and PI3K-inhibitors, but also by the CaMKII inhibitor KN93. THP-1 and SF macrophages produced high levels of VEGF, IL-8, and MMP-9, and VEGF protein production was significantly inhibited by PI3K-inhibitor, and by both CaMKII inhibitors. LPS stimulation in an hypoxic environment did not change VEGF levels, suggesting that LPS induced VEGF production in macrophages is more important than the hypoxic induction.</p> <p>Conclusions</p> <p>Expression of HIF-1 alpha and downstream effects in macrophages are regulated by ERK-, PI3K, but also by CaMKII pathways. Inhibition of HIF-1α protein expression and significant inhibition of VEGF production in macrophages was found using CaMKII inhibitors. This is an unknown but very interesting effect of the CaMKII inhibitor SMP-114, which has been in clinical trial as DMARD for the treatment of RA. This effect may contribute to the anti-arthritic effects of SMP-114.</p

Standardised assessment of membrane proteinase 3 expression. Analysis in ANCA‐associated vasculitis and controls

Objectives: Increased numbers of neutrophils expressing proteinase 3 on their membrane (mPR3) have been reported in anti-neutrophil cytoplasm antibody ( ANCA)-associated vasculitis (AAV) and are suggested to be involved in AAV immunopathogenesis. In most studies, neutrophils were analysed for mPR3 expression without priming with TNF alpha, suggesting that mPR3 expression on neutrophils is dependent on other priming events, such as isolation procedures. These priming events can be variable. Therefore, we analysed mPR3 expression on neutrophils before and after priming with TNFa to assess whether standardised assessment of mPR3 expression requires priming. Using neutrophils before and after priming with TNFa, we assessed percentages of mPR(3+) neutrophils in patients with AAV and in disease and healthy controls. Methods: Neutrophils from patients with PR3-AAV and MPO-AAV, systemic lupus erythematosus (SLE) and rheumatoid arthritis ( RA), and from healthy controls were analysed before and after priming with TNFa for mPR3 expression. Results: 42% of all individuals analysed showed minimal expression for mPR3 on all neutrophils before priming with TNFa, whereas after priming a clear mPR3+ subset was observed next to mPR(3-) neutrophils, corresponding to bimodal mPR3 expression. In patients with PR3-AAV or MPO-AAV, the percentage of mPR3+ neutrophils after priming with TNFa was significantly increased ( p <0.01 and p, 0.05, respectively) compared with healthy controls. Percentages of mPR3+ PMN were also increased in patients with SLE ( p <0.01) but not in RA. Conclusion: Standardised assessment of proteinase 3 on the membrane of neutrophils requires priming with TNFa. Percentages of mPR3+ PMN are increased in AAV and SLE, but not in RA

The effectiveness and acceptability of different cueing devices for people with parkinsons disease and gait initiation difficulties

Purpose: To investigate the effects of three cueing devices (visual, auditory, and somatosensory) on movement and muscular control during gait initiation in people with Parkinson’s disease who experience freezing and to ascertain the acceptability of these cueing devices to novice users. Relevance: Over six million people worldwide are affected by Parkinson’s disease. It is estimated that thirty percent may experience freezing (an inability to produce effective steps) and have difficulty initiating gait, and maintaining gait whilst negotiating obstacles. This limits their capacity for physical activity and ability to participate in usual activities, and can result in social isolation. Various sensory cues have been shown to improve on-going gait in people with Parkinson’s Disease, but there is limited evidence on their effectiveness for people with gait initiation difficulties. Jiang and Norman (2006) found that transverse line visual cues improved gait initiation, while auditory cues had no effect. Dibble et al. (2004), using auditory and cutaneous cues during maximal speed gait initiation found an adverse effect on movement outcomes. Portable cueing devices are commercially available, however their influence in gait initiation and acceptability to users is currently unknown. InformaWPT2011, Research Report Abstracts eS773 tion on their effectiveness would enable physical therapists to provide better informed advice to potential purchasers. Participants: Twenty participants with idiopathic Parkinson’s disease and a history of freezing of gait (evaluated using item 14 of the Unified Parkinson’s Disease Rating Scale) were recruited; 14 males and 6 females, mean age 68 years and 11.5 years since diagnosis. Methods: An experimental trial of five randomised conditions: laser cane, sound metronome, vibrating metronome, walking stick and uncued. After using each cue participants’ opinions were obtained via a questionnaire. Motion data were collected using a 10 camera motion analysis system, force platforms and surface Electromyography. Analysis: Questionnaire responses from twelve participants who experienced freezing during testing were analysed using a Wilcoxon signed ranks test. Motion data from these participants were analysed using one-way ANOVA tests with post-hoc pair-wise comparisons to test for differences between conditions. Results: Significant differences were seen in step length, Centre of Mass and Centre of Pressure movement in the anterior/ posterior and medial/lateral directions between freezing and non-freezing episodes. The post hoc pair-wise comparisons showed significant improvements in the Centre of Mass and Centre of Pressure movement when using the laser cane and the walking stick and greater step length when using the laser cane. Participants rated the perceived effectiveness of the devices, theWilcoxon test showed a significant improvement in satisfaction when using the laser cane for both starting and maintaining walking (p < 0.05). Conclusions: The laser cane was overall the most effective cueing device for people with Parkinson’s disease and gait initiation difficulties in both user’s perception and efficacy tested in the gait laboratory. However the longer term effectiveness and acceptability of cueing devices at home and outdoors requires further investigation. Implications: This study would support the use of the laser cane as a relatively cheap intervention for people with Parkinson’s disease who experience spontaneous freezing

Time course of induction of mRNA expression of tumor necrosis factor (TNF)-α, IL-1β, IL-6, IL-8, and cyclooxygenase 2 (COX-2)

<p><b>Copyright information:</b></p><p>Taken from "Strong inhibition of TNF-α production and inhibition of IL-8 and COX-2 mRNA expression in monocyte-derived macrophages by RWJ 67657, a p38 mitogen-activated protein kinase (MAPK) inhibitor"</p><p>Arthritis Research & Therapy 2004;6(4):R384-R392.</p><p>Published online 21 Jun 2004</p><p>PMCID:PMC464924.</p><p>Copyright © 2004 Westra et al.; licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.</p> Monocyte-derived macrophages from healthy controls (= 2) were stimulated for increasing periods of time with lipopolysaccharide (50 ng/ml). mRNA expression was determined with real-time RT-PCR and results were calculated as fold induction in comparison with unstimulated cells (fold induction = 1)

Humoral responses after influenza vaccination are severely reduced in patients with rheumatoid arthritis treated with rituximab

Objective. For patients with rheumatoid arthritis (RA), yearly influenza vaccination is recommended. However, its efficacy in patients treated with rituximab is unknown. The objectives of this study were to investigate the efficacy of influenza vaccination in RA patients treated with rituximab and to investigate the duration of the possible suppression of the humoral immune response following rituximab treatment. We also undertook to assess the safety of influenza vaccination and the effects of previous influenza vaccination. Methods. Trivalent influenza subunit vaccine was administered to 23 RA patients who had received rituximab (4-8 weeks after rituximab for 11 patients [the early rituximab subgroup] and 6-10 months after rituximab for 12 patients [the late rituximab subgroup]), 20 RA patients receiving methotrexate (MTX), and 29 healthy controls. Levels of antibodies against the 3 vaccine strains were measured before and 28 days after vaccination using hemagglutination inhibition assay. The Disease Activity Score in 28 joints (DAS28) was used to assess RA activity. Results. Following vaccination, geometric mean titers (GMTs) of antiinfluenza antibodies significantly increased for all influenza strains in the MTX-treated group and in healthy controls, but for no strains in the rituximab-treated group. However, in the late rituximab subgroup, a rise in GMT for the A/H3N2 and A/H1N1 strains was demonstrated, in the absence of a repopulation of CD19+ cells at the time of vaccination. Seroconversion and seroprotection occurred less often in the rituximab-treated group than in the MTX-treated group for the A/H3N2 and A/H1N1 strains, while seroprotection occurred less often in the rituximab-treated group than in the healthy controls for the A/H1N1 strain. Compared with unvaccinated patients in the rituximab-treated group, previously vaccinated patients in the rituximab-treated group had higher pre- and postvaccination GMTs for the A/H1N1 strain. The DAS28 did not change after vaccination. Conclusion. Rituximab reduces humoral responses following influenza vaccination in RA patients, with a modestly restored response 6-10 months after rituximab administration. Previous influenza vaccination in rituximab-treated patients increases pre- and postvaccination titers. RA activity was not influenced

Novel Association in Chromosome 4q27 Region with Rheumatoid Arthritis and Confirmation of Type 1 Diabetes Point to a General Risk Locus for Autoimmune Diseases

Recently, association of celiac disease with common single-nucleotide polymorphism (SNP) variants in an extensive linkage-disequilibrium block of 480 kb containing the KIAA1109, Tenr, IL2, and IL21 genes has been demonstrated in three independent populations (rs6822844 Pcombined=1.3×10-14). The KIAA1109/Tenr/IL2/IL21 block corresponds to the Idd3 locus in the nonobese diabetic mouse model of type 1 diabetes (T1D). This block was recently found to be associated with T1D in a genomewide association study, although this finding lacks unequivocal confirmation. We therefore aimed to investigate whether the KIAA1109/Tenr/IL2/IL21 region is involved in susceptibility to multiple autoimmune diseases. We tested SNP rs6822844 for association with disease in 350 T1D-affected and 1,047 rheumatoid arthritis (RA)–affected Dutch patients and in 929 controls. We replicated the association with T1D (P=.0006; OR 0.64 [95% CI 0.50–0.83]), and revealed a similar novel association with RA (P=.0002; OR 0.72 [95% CI 0.61–0.86]). Our results replicate and extend the association found in the KIAA1109/Tenr/IL2/IL21 gene region with autoimmune diseases, implying that this locus is a general risk factor for multiple autoimmune diseases