Objective. Immune-mediated inflammatory arthritis (IMIA) is a heterogeneous group of diseases including rheumatoid arthritis (RA), psoriatic arthritis (PsA), and spondyloarthritis (SpA). Disease-modifying antirheumatic drugs (DMARDs) target very different cellular components of the disease processes. Characterization of the pathobiological subtypes of IMIA could provide more specific treatment approaches for each disease. For example, RA has been proposed to consist of at least three synovial pathotypes (lymphoid, myeloid, and fibroid), and only a subgroup of RA patients have erosive disease. The objective of this study was to evaluate the effects of various DMARDs on different synovial cell subsets using human ex vivo models of IMIA.Methods. Synovial fluid and blood samples were obtained from a study population consisting of patients with RA, PsA, or peripheral SpA with at least one swollen joint (n = 18). The DMARDs used in this study were methotrexate, adalimumab, etanercept, tocilizumab, anakinra, ustekinumab, secukinumab, tofacitinib, and baricitinib. Paired synovial fluid mononuclear cells (SFMCs), peripheral blood mononuclear cells (PBMCs), and fibroblast-like synovial cells (FLSs) were used in three different previously optimized ex vivo models.Results. In SFMCs cultured for 48 hours, all DMARDs except anakinra decreased the production of monocyte chemoattractant protein (MCP)-1. In SFMCs cultured for 21 days, only the two tumor necrosis factor alpha (TNFα) inhibitors adalimumab and etanercept decreased the secretion of tartrate-resistant acid phosphatase (P < 0.01, P < 0.001). In the FLS and PBMC 48-hour co-cultures, only tocilizumab (P < 0.001) and the two Janus kinase inhibitors tofacitinib and baricitinib (both P < 0.05) decreased the production of MCP-1 by around 50%.Conclusion. TNFα inhibition was effective in preventing inflammatory osteoclastogenesis, whereas tocilizumab, tofacitinib, and baricitinib had superior efficacy in cultures dominated by FLSs. Taken together, this study reveals that responses to cytokine inhibitors associate with cellular composition in models of IMIA. In particular, this study provides new evidence on the differential effect of DMARDs on leukocytes compared with stromal cells.

Objective Patients accommodated in single-bed rooms may have a reduced risk of hospital-acquired infections (HAIs) compared to those in multi-bed rooms. This study aimed to examine the effect of single-bed accommodation on HAIs in older patients admitted to a geriatric ward. Methods A retrospective cohort study of patients admitted to geriatric wards in a university hospital in Central Denmark Region linked to a move to a newly built hospital, involving all consecutively admitted patients aged 65 years and over from 15 September to 19 December 2016 and a similar cohort admitted in the same three months in 2017. We compared the incidence of HAIs in patients in single-bed accommodation to those in multi-bed accommodation using retrospective review of electronic patient records, with all infections verified microbiologically or by X-ray with onset between 48 hours after admission to 48 hours after discharge from hospital. Results In total 446 patients were included. The incidence of HAIs in multi-bed accommodation was 30% compared to 20% in single-bed accommodation. The hazard ratio was 0.62 (95% Confidence Interval 0.43–0.91, p = 0.01) for single-bed accommodation. This finding remained robust after adjustment for age, sex, infection at admission, risk of sepsis, use of catheter, treatment with prednisone or methotrexate, and comorbidity index. Conclusion Accommodation in single-bed rooms appeared to reduce HAIs compared to multi-bed rooms in two geriatric wards. This finding should be considered as hypothesis-generating and be examined further using an experimental design.

Background Resveratrol is a natural polyphenol found in berries, roots and wine that is well known to have anti-inflammatory and anti-oxidative properties. The anti-inflammatory effect has been reported for both immune cells and connective tissues, but only few studies have investigated effects on immune mediated inflammatory arthritis. None of which have studied this effect when combining resveratrol with methotrexate or adalimumab, two major drugs in the treatment of immune mediated inflammatory arthritis. We therefore aimed to investigate the anti-inflammatory effect of resveratrol alone and in combination with methotrexate or adalimumab in ex vivo models of immune mediated inflammatory arthritis. We furthermore aimed to describe any variations in this effect based on disease activity and cellular composition of the synovial fluid infiltrate. Methods Synovial fluid mononuclear cells from patients with rheumatoid arthritis ( n = 7) and spondyloarthritis ( n = 7) were cultured for either 48 h or 21 days. In both models, synovial fluid mononuclear cells were treated with resveratrol alone or in combination with methotrexate or adalimumab. Monocyte chemoattractant protein 1, matrix metalloproteinase 3 and tartrate resistant acidic phosphatase were measured to quantify inflammation, enzymatic degradation and osteoclast differentiation, respectively. Results Resveratrol reduced monocyte chemoattractant protein 1 production by synovial fluid mononuclear cells significantly ( p = 0.005) compared to untreated controls. The effect of resveratrol was greatest in cultures from patients with low disease activity, i.e. DAS28CRP ≤ 3.2 ( p = 0.022), and in cultures dominated by lymphocytes ( p = 0.03). Further, the combination of methotrexate and resveratrol significantly reduced monocyte chemoattractant protein 1 levels compared with methotrexate alone in cultures from patients with low disease activity ( p = 0.016), and in cultures with high lymphocyte count ( p = 0.011). Resveratrol did not significantly affect matrix metalloproteinase 3 and tartrate resistant acidic phosphatase production. Conclusion Resveratrol has anti-inflammatory properties in our ex vivo model of immune mediated inflammatory arthritis. Results show an additive effect of resveratrol, when combined with methotrexate in samples dominated by lymphocytes and samples from patients with low disease activity. This suggests further investigations in vitro and whether this effect may also be present in a clinical setting. Electronic supplementary material The online version of this article (10.1186/s41927-018-0036-5) contains supplementary material, which is available to authorized users.

BackgroundResveratrol (RSV), a non-toxic polyphenol found in grapes, certain nuts, roots etc., have received increased attention in the last decade due to its anti-inflammatory modulation of a number of pathways, including cyclooxygenase-1/-2, nuclear factor kappa-beta and cytokine production. In vitro, RSV has been shown to reduce production of interleukin 1-beta and tumor necrosis factor alpha in monocytes and inhibit T-cell activation and synoviocyte proliferation. In vivo, intra-articular injections of RSV have demonstrated anti-inflammatory and pannus inhibiting effects in rats with induced arthritis.ObjectivesHere, we tested whether the anti-inflammatory effect of RSV in arthritis patients depends on the degree of systemic inflammation and the cellular composition of extracted synovial fluid. Furthermore, we evaluated the anti-inflammatory effect of RSV in combination with methotrexate (MTX) and adalimumab.MethodsSynovial fluid mononuclear cells (SFMCs) from patient with rheumatoid arthritis (n=7) and spondyloarthritis (n=7) were cultured in monoculture for 48 hours (in vivo activated lymphocytes and monocytes) or 21 days (spontaneous generation of osteoclasts). Cultures were either left untreated or treated with RSV (25 μM), methotrexate (0.5 μg/ml), adalimumab (5 μ g/ml) or in combination. Supernatants were analysed for the production of monocyte chemoattractant protein-1 (MCP-1) and matrix metalloproteinase 3 (MMP3). Osteoclast differentiation was analysed with a tartrate resistant acidic phosphatase (TRAP) enzymatic assay.ResultsIn the SFMC cultures, RSV reduced MCP-1 production by 25% compared with untreated cells (P=0.032). When grouping results by c-reactive protein (CRP), i.e. < median vs. ≥ median, the inhibitory effect of RSV was primarily seen in cultures from patients with CRP <21 mg/l. In this group, RSV reduced MCP-1 production by 63%. Combining MTX and RSV reduced MCP-1 production compared to MTX alone, but only in the group of patients with CRP <21 mg/l (P=0.002). Combining adalimumab and RSV seemed to reduce MCP-1 in the group of patients with CRP <21 mg/l, but increase production in patients with CRP ≥21 mg/l (P=0.03). Similar grouping based on lymphocyte count showed RSV, MTX and adalimumab, alone or in combination, all reduced MCP-1 significantly compared to untreated cells in cultures from patients with ≥62% lymphocytes in the synovial fluid. RSV, MTX and adalimumab did not affect MMP3 production in the SFMC cultures. In the osteoclast cultures, RSV alone did not affect MCP-1 or TRAP. However, the combination of RSV and MTX reduced MCP-1 compared to no treatment (P=0.004). Adalimumab alone or combined with RSV reduced TRAP compared with untreated cultures (P<0.027).ConclusionsRSV exhibits an anti-inflammatory effect on SFMCs. Interestingly; our data suggest that this effect is most pronounced in patients with relatively low CRP. Further, RSV produces an additive anti-inflammatory response in combination with MTX in the group of patients with low CRP and a synovial fluid dominated by lymp...

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BackgroundTargeting intracellular pathways with oral small molecules is an attractive therapeutic approach for treating immune mediated inflammatory diseases. The mitogen-activated protein kinase (MAPK) pathway is activated by environmental stressors, growth factors and inflammatory cytokines. However, the inhibition of central MAPK proteins has so far had undesirable side effects. The MAPK-activated protein kinase 2 (MK2) is a downstream mediator in the MAPK signalling pathway and could therefore be inhibited without the same side effects (see figure 1).ObjectivesThe objective of this study was to study the effects of a small molecule inhibiting MK2 on inflammation and structural changes in ex vivo models of immune mediated inflammatory arthritis.MethodsSynovial fluid mononuclear cells (SFMCs), fibroblast like synovial cells (FLSs) and peripheral blood mononuclear cells (PBMCs) were obtained from a study population consisting of patients with active RA or peripheral SpA with at least one swollen joint (for obtaining synovial fluid) (n=14). SFMCs were cultured for 48 hours with and without addition of a MK2 inhibitor (Celgene) at 1000 nM, 333 nM and 111 nM and supernatants were analysed by the Olink proseek multiplex interferon panel and commercially available ELISA assays. Because FLSs are only found in small amounts among SFMCs, autologous co-cultures of FLS and PBMCs and SFMCs were also used. SFMCs cultured for 21 days were used to study inflammatory macrophage differentiation and osteoclastogenesis.ResultsIn SFMCs cultured for 48 hours, the MK2 inhibitor decreased the production of CXCL9 (p<0.001), CXCL10 (p<0.01), HGF (p<0.01), CXCL11 (p<0.01), TWEAK (p<0.05), and IL-12B (p<0.05) and increased the production of CXCL5 (p<0.0001), CXCL1 (p<0.0001), CXCL6 (p<0.001), TGFα (p=0.01), MCP-3 (p<0.01), LAP TGFβ (p<0.05) dose-dependently after Bonferroni correction (all corrected P values). At the highest concentration, the MK2 inhibitor also decreased MCP-1 production (p<0.05). In FLS-SFMC co-cultures, the MK2 inhibitor decreased MCP-1 production (p<0.05) but did not change the production of DKK1 and MMP3. In FLS-PBMC co-cultures, the MK2 inhibitor decreased the production of MCP-1 (p<0.0001), increased MMP3 production (p<0.05) but did not change DKK1 production. In SFMCs cultured for 21 days as a model of inflammatory macrophage differentiation and osteoclastogenesis, the MK2 inhibitor decreased the production of MCP-1 (p<0.05) and tartrate-resistant acid phosphatase (TRAP) (p<0.05) but did not change the production of IL-10.Abstract THU0096 – Figure 1Modified from Wagner & Nebreda, Nature Review Cancer, 2009.ConclusionsThis study reveals the effects of a MK2 inhibitor in ex vivo models of immune mediated inflammatory arthritis. The MK2 inhibitor changed the secretory profile of SFMCs and decreased inflammatory osteoclastogenesis. Taken together, this points to a role of this MK2 inhibitor in attenuating inflammatory and destructive arthritis.Disclosure of InterestT. Kragstrup: None declared, A. Mellemkjær: None decl...