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ObjectiveAs part of European League against Rheumatism (EULAR)/European Musculoskeletal Conditions Surveillance and Information Network, 20 user-focused standards of care (SoCs) for rheumatoid arthritis (RA) addressing 16 domains of care were developed. This study aimed to explore gaps in implementation of these SoCs across Europe.MethodsTwo cross-sectional surveys on the importance, level of and barriers (patients only) to implementation of each SoC (0–10, 10 highest) were designed to be conducted among patients and rheumatologists in 50 European countries. Care gaps were calculated as the difference between the actual and maximum possible score for implementation (ie, 10) multiplied by the care importance score, resulting in care gaps (0–100, maximal gap). Factors associated with the problematic care gaps (ie, gap≥30 and importance≥6 and implementationResultsOverall, 26 and 31 countries provided data from 1873 patients and 1131 rheumatologists, respectively. 19 out of 20 SoCs were problematic from the perspectives of more than 20% of patients, while this was true for only 10 SoCs for rheumatologists. Rheumatologists in countries with lower gross domestic product and non-European Union countries were more likely to report problematic gaps in 15 of 20 SoCs, while virtually no differences were observed among patients. Lack of relevance of some SoCs (71%) and limited time of professionals (66%) were the most frequent implementation barriers identified by patients.ConclusionsMany problematic gaps were reported across several essential aspects of RA care. More efforts need to be devoted to implementation of EULAR SoCs.

Objective.To determine whether serum trough concentrations of tocilizumab (TCZ) administered as a fixed-dose subcutaneous (SC) injection for the treatment of rheumatoid arthritis (RA) are associated with disease activity responses.Methods.We analyzed datasets from the Israeli branch of the multinational TOZURA study, which evaluated a weekly subcutaneous TCZ treatment regimen in a real-life clinical setting. Generalized estimating equations (GEE) were used to evaluate associations between the TCZ levels and the study outcomes. Linear models and GEE were used to evaluate associations between patient characteristics and TCZ levels.Results.A significant association between the TCZ concentrations and the change in the Clinical Disease Activity Index (CDAI) score was observed. In a multivariate binary GEE model, every increase of 10 µg/ml in the concentration of TCZ was associated with being in a state of CDAI remission or low disease activity (OR 1.41) versus a moderate/high disease activity state. An OR of 1.52 was associated with being in a state of Health Assessment Questionnaire–Disability Index remission. In univariate linear models, there was an inverse association between body mass index (BMI) and improvement in the CDAI score, and the BMI score was associated with lower TCZ concentrations. Patients who weighed > 100 kg had lower TCZ concentrations.Conclusion.In the first 24 weeks of treatment with SC TCZ injections, TCZ concentrations were associated with clinical improvement, while body weight and BMI were inversely associated with TCZ concentrations. Personalizing the dose of SC TCZ to body weight may improve outcomes of clinical disease activity in patients with RA.

Nucleotide pyrophosphatase/phosphodiesterase-1 (NPP1) inhibitors have been suggested as a potential treatment for calcium pyrophosphate dihydrate (CPPD) deposition disease. Here, we targeted the development of improved NPP1 inhibitors based on acyclic mimics of Pα,α-phosphorodithioate-substituted adenine nucleotides, 7-10. The latter were obtained in a facile two-step synthesis from adenine-(methoxy)ethanol. Among analogs 7-10, adenine-(methoxy)ethoxy-Pα,α-dithio-triphosphate, 8, was the most potent NPP1 inhibitor both with purified enzyme (IC50 0.645 μM) and in osteoarthritic human chondrocytes (IC50 0.033 μM). Furthermore, it efficaciously (10-fold vs. control) inhibited ATP-induced CPPD in human articular chondrocytes. Importantly, 8 was a highly selective NPP1 inhibitor which showed only minor inhibition of NPP3, CD39 and CD73, and did not inhibit TNAP (tissue nonspecific alkaline phosphatase) activity in human chondrocytes. Furthermore, 8 did not activate P2Y1,2,6 receptors. Analog 8 was not toxic to cultured chondrocytes at 100 μM. Therefore, 8 may be suitable for further development as a drug candidate for the treatment of CPPD arthritis and other NPP1-related diseases.

Objective Subclinical myocardial dysfunction has been reported to occur early in systemic lupus erythematous (SLE). The study aim was to search for biomarkers of subclinical myocardial dysfunction which may correlate with disease activity in SLE patients. Methods This is a prospective, controlled, cross-sectional study of 57 consecutive patients with SLE and 18 controls. Serum samples were obtained to determine serum soluble ST2 (sST2), CXCL-10 and high-sensitivity troponin (hs-troponin) levels. All participants underwent an echocardiographic tissue Doppler study. Results sST2, CXCL-10 and hs-troponin levels were higher in patients with higher SLE disease activity (SLEDAI). sST2 and CXCL-10 levels were higher in patients with more disease damage as measured by the SLE damage index. Measures of diastolic dysfunction, as assessed by echocardiographic tissue Doppler negatively correlated with log CXCL-10: including E/A; E/e′lateral and E/e′septal, while E/e′ positively correlated with CXCL-10. Diastolic dysfunction parameters also correlated with log sST2 levels, a negative correlation was seen with E/e′lateral and a positive correlation was seen with E/e′. Systolic dysfunction parameters positively correlated with hs-troponin: LVED, LVES, IVS, LVMASS and LVMASS index. In a multivariate analysis, sST2 and CXCL-10 were found to be significantly different in SLE vs. healthy controls, independent of each other and independent of cardiovascular risk factors. Conclusions Soluble ST2 and CXCL-10 are markers of disease activity and accrued damage in SLE and may serve as sensitive biomarkers for detection of subclinical diastolic dysfunction, independent of traditional cardiovascular risk factors.

Summary Objectives The chondrocytes' pericellular matrix acts as a mechanosensor by sequestering growth factors that are bound to heparan sulfate (HS) proteoglycans. Heparanase is the sole mammalian enzyme with HS degrading endoglycosidase activity. Here, we aimed to ascertain whether heparanase plays a role in modulating the anabolic or catabolic responses of human articular chondrocytes. Methods Primary chondrocytes were incubated with pro-heparanase and catabolic and anabolic gene expression was analyzed by quantitative polymerase chain reaction (PCR). MMP13 enzymatic activity in the culture medium was measured with a specific fluorescent assay. Extracellular regulated kinase (ERK) phosphorylation was evaluated by Western blot. Human osteoarthritis (OA) cartilage was assessed for heparanase expression by reverse-transcriptase PCR, by Western blot and by a heparanase enzymatic activity assay. Results Cultured chondrocytes rapidly associated with and activated pro-heparanase. Heparanase induced the catabolic genes MMP13 and ADAMTS4 and the secretion of active MMP13, and down-regulated the anabolic genes ACAN and COL2A1. PG545, a HS-mimetic, inhibited the effects of heparanase. Heparanase expression and enzymatic activity were demonstrated in adult human osteoarthritic cartilage. Heparanase induced ERK phosphorylation in cultured chondrocytes and this could be inhibited by PG545, by fibroblast growth factor 2 (FGF2) neutralizing antibodies and by a FGF-receptor inhibitor. Conclusions Heparanase is active in osteoarthritic cartilage and induces catabolic responses in primary human chondrocytes. This response is due, at least in part, to the release of soluble growth factors such as FGF2.

Nucleotide pyrophosphatase/phosphodiesterase-1 (NPP1) inhibitors have been suggested as a potential treatment for calcium pyrophosphate dihydrate (CPPD) deposition disease. Here, we targeted the development of improved NPP1 inhibitors based on acyclic mimics of Pα,α-phosphorodithioate-substituted adenine nucleotides, 7-10. The latter were obtained in a facile two-step synthesis from adenine-(methoxy)ethanol. Among analogs 7-10, adenine-(methoxy)ethoxy-Pα,α-dithio-triphosphate, 8, was the most potent NPP1 inhibitor both with purified enzyme (IC50 0.645 μM) and in osteoarthritic human chondrocytes (IC50 0.033 μM). Furthermore, it efficaciously (10-fold vs. control) inhibited ATP-induced CPPD in human articular chondrocytes. Importantly, 8 was a highly selective NPP1 inhibitor which showed only minor inhibition of NPP3, CD39 and CD73, and did not inhibit TNAP (tissue nonspecific alkaline phosphatase) activity in human chondrocytes. Furthermore, 8 did not activate P2Y1,2,6 receptors. Analog 8 was not toxic to cultured chondrocytes at 100 μM. Therefore, 8 may be suitable for further development as a drug candidate for the treatment of CPPD arthritis and other NPP1-related diseases.

Objectives Calcium pyrophosphate deposition (CPPD) is associated with osteoarthritis and is the cause of a common inflammatory articular disease. Ecto-nucleotide pyrophosphatase/phosphodiesterase 1 (eNPP1) is the major ecto-pyrophosphatase in chondrocytes and cartilage-derived matrix vesicles (MVs). Thus, eNPP1 is a principle contributor to extracellular pyrophosphate levels and a potential target for interventions aimed at preventing CPPD. Recently, we synthesized and described a novel eNPP1-specific inhibitor, SK4A, and we set out to evaluate whether this inhibitor attenuates nucleotide pyrophosphatase activity in human OA cartilage. Methods Cartilage tissue, chondrocytes and cartilage-derived MVs were obtained from donors with OA undergoing arthroplasty. The effect of SK4A on cell viability was assayed by the XTT method. eNPP1 expression was evaluated by western blot. Nucleotide pyrophosphatase activity was measured by a colorimetric assay and by HPLC analysis of adenosine triphosphate (ATP) levels. ATP-induced calcium deposition in cultured chondrocytes was visualized and quantified with Alizarin red S staining. Results OA chondrocytes expressed eNPP1 in early passages, but this expression was subsequently lost upon further passaging. Similarly, significant nucleotide pyrophosphatase activity was only detected in early-passage chondrocytes. The eNPP1 inhibitor, SK4A, was not toxic to chondrocytes and stable in culture medium and human plasma. SK4A effectively inhibited nucleotide pyrophosphatase activity in whole cartilage tissue, in chondrocytes and in cartilage-derived MVs and reduced ATP-induced CPPD. Conclusion Nucleotide analogues such as SK4A may be developed as potent and specific inhibitors of eNPP1 for the purpose of lowering extracellular pyrophosphate levels in human cartilage with the aim of preventing and treating CPPD disease.

Diffuse idiopathic skeletal hyperostosis (DISH) is a non-inflammatory condition that involves calcification and ossification of the spinal ligaments and entheses. While, characteristic magnetic resonance imaging (MRI) lesions of the spine in patients with axial spondyloarthritis, another enthesitis-related disease, have been described and defined, there is a paucity of information regarding the MRI findings in DISH. The aim of this study was to describe the MRI findings of patients with DISH. We collected computed tomography studies with findings characteristic of DISH and that also had corresponding and concurrent MRI studies of the spine. For each patient, sagittal T1-weighted and STIR MRI sequences were evaluated for anterior/posterior vertebral corners of bone marrow edema (BME) and fat deposition. In total, we assessed 156 vertebral units in 10 patients that had both radiographic evidence of DISH and available MRI studies of the spine. Lesions consistent with BME corners were detected in five patients, and in three of them, three separate sites were involved, a finding that is suggestive of axial spondyloarthritis (SpA) according to the ASAS/OMERACT consensus statement. Fat deposition corners were detected in eight patients and in seven of them, several sites were involved. Spinal MRI lesions that are characteristic of axial SpA were commonly observed in a cohort of patients with DISH. This bears relevance to cases with diagnostic uncertainty and may imply overlapping pathogenetic mechanisms for new bone formation in both SpA and DISH. Further study is indicated to better characterize the similarities and differences between the MRI lesions of DISH and SpA.

Patients with psoriatic arthritis (PsA) are at increased risk of cardiovascular disease (CVD). High-sensitivity cardiac troponin T (hs-cTnT) is a novel biomarker of CVD. The objective of this study is to determine the prevalence of circulating hs-cTnT in patients with PsA compared to the general population and to characterize a PsA subset with detectable hs-cTnT. A cross-sectional analysis of serum hs-cTnT levels was performed in 116 consecutive patients with PsA and the Tel-Aviv Medical Center Inflammatory Survey cohort of the general population (n = 6052) as a control group. The level and prevalence of hs-cTnT (ng/L) were similar in the entire study population: 4.94 ± 4.4, 30.2% in PsA, 5.17 ± 6.7, 34.2% and 5.38 ± 4.3, 37.9% in unmatched and matched control groups according to age, gender and cardiovascular risk factors, respectively. Factors associated with detectable hs-cTnT in PsA included male gender (p = 0.002), age (p = 0.007), hypertension (p

Objective To assess the immunogenicity and safety of vaccination against seasonal influenza in psoriatic arthritis (PsA) patients treated with secukinumab versus healthy controls (HC). Methods PsA patients administered secukinumab for ≥3 months and HC received the Sanofi Pasteur vaccine composed of 3 antigens (H3N3, H1N1, and B) and underwent clinical and laboratory assessments on the day of vaccination and 4–6 weeks later. Immunogenicity of the vaccine was evaluated by hemagglutination inhibition assay against those 3 antigens. Responders to each antigen were defined by a 4-fold increase in the antigen titer or by seroconversion in patients whose baseline level was Results Thirty-two consecutive PsA patients treated with secukinumab for ≥3 months comprised the study group, 10 of whom received concomitant conventional synthetic disease-modifying drugs, mostly methotrexate. There were 17 age- and gender-matched HC (median age 48.5 years, 6 females). The geometric mean titers of each antigen increased significantly in both groups. The number of responders in each group was similar for H3N2 and H1N1, and significantly higher for B/Brisbane in the PsA group. The proportion of patients with a seroprotective level (a titer >1/40) was high and similar in both groups. There was no correlation between the response rate and age, gender, or selected parameters of disease activity (tender/swollen joint counts, Leeds enthesitis index, physician and patient global assessment, psoriasis area severity index, and C-reactive protein). No disease exacerbation was observed following the vaccination. No serious adverse effects were observed in both groups during the study period. Conclusion Secukinumab treatment does not affect the humoral response to influenza vaccine of patients with PsA.

Overexpression of ecto-nucleotide pyrophosphatase-1 (NPP1) is associated with diseases such as calcium pyrophosphate dihydrate deposition disease, calcific aortic valve disease, and type 2 diabetes. In this context, NPP1 inhibitors are potential drug candidates for the treatment of these diseases. The present study focuses on the analysis of the structure-activity relationship of NPP1 inhibitors based on acyclic uracil-nucleotides. For this purpose, we synthesized acyclic uridine-monophosphate analogs, 10-11, uridine-diphosphate analogs, 12–14, and uridine-Pα,α-dithio-triphosphate analogs, 15–17. We evaluated their inhibitory activity and selectivity towards NPP1, -3, NTPDase1, -2, -3, and -8, and P2Y2,4,6 receptors. Analogs 16 and 17 were the most selective and potent NPP1 inhibitors (Ki 0.94 and 0.73 μM, respectively) among the tested molecules. Analogs 10–17 had only minute effect on uracil-nucleotide responding P2Y2,4,6 receptors. Analog 17 (100 μM) displayed 96% inhibition of NPPase activity in osteoarthritic human chondrocytes. Analogs 14–17 displayed weak inhibitory effect on alkaline phosphatase activity at equimolar concentrations in human chondrocytes. All tested analogs showed no toxicity at human chondrocytes. We concluded that ribose-ring to chain transformation, as well as the type of the nucleobase, are parameters of minor significance to NPP1 inhibition, whereas the major parameter is Pα-dithio-substitution. In addition, the length of the phosphate chain also significantly affects inhibition. Overall, the experimental results were well reproduced by molecular docking. A correlation was observed between the activities of the compounds and the number of H-bonds and salt bridges formed between the inhibitors and NPP1 binding site residues. Uracil-N1-(methoxy)ethyl-β-Pα,α-dithio, Pβ,γ-methylene tri-phosphate, 17, was identified as the most potent, selective, and non-toxic NPP1 inhibitor among the tested analogs, and may be used as a lead structure for further drug development.

Background Serum trough levels of TNFα inhibitor biologics have been found to be associated with clinical responses in rheumatoid arthritis (RA) patients. It is unknown whether serum trough levels of tocilizumab (TCZ) administered as a fixed dose subcutaneous injection, also associate with clinical disease activity responses. Objectives To ascertain whether serum TCZ trough levels at weeks 12 and 24 after beginning treatment with a 162 mg once weekly regimen of SC tocilizumab, were associated with clinical disease activity outcomes in RA patients. Methods We analysed data sets from the Israeli branch (TASC, NCT01988012) of the Roche multinational umbrella study TOZURA, which evaluated a SC TCZ treatment regimen of 162 mg once weekly as monotherapy or in combination with methotrexate or other csDMARDs in a real-life clinical setting. The study comprised of 100 patients. A paired-samples T test was used to compare mean serum TCZ levels at week 12 relative to week 24. Clinical disease activity index (CDAI) scores were natural log-transformed in order to achieve normal distribution. Generalised estimating equations were used to evaluate associations between the predictors (TCZ levels, soluble IL-6 receptor (sIL6R) levels) and the study outcomes (CDAI scores, HAQ scores, CDAI remission/low disease activity status, HAQ DI remission). Generalised estimating equations were also used to evaluate associations between age, sex, weight, BMI, baseline CRP levels, serum TCZ and sIL6r serum levels. P values below 0.05 were considered significant. Results Serum trough levels of TCZ at week 24 (mean 41.1, SD 23.2) were higher than at week 12 (mean 36.3, SD 18.1). In a univariate analysis, for every increase of 1 microgram/ml in the serum concentration of TCZ there was a corresponding decrease of 1.3% (95% CI: 0.4% to 2.3%) in the CDAI score and for every increase of 100 ng/ml in the serum concentration of sIL6R there was a corresponding decrease of 12.6% (95% CI: 2% to 22%) in the CDAI score. In a multivariate model which included age, sex, visit date and both sIL6R and TCZ levels, only the associated between TCZ levels and CDAI scores remained significant. Similarly, every increase of 10 microgram/ml in the serum concentration of TCZ was associated with an odds ratio of 1.35 (95% CI: 1.07 to 1.72) of being in a state of CDAI remission or low disease activity versus moderate/high disease activity state. TCZ and sIL6R serum levels were not associated with HAQ DI scores. Female sex was associated with an increase of 12.9 microgram/ml in the serum TCZ concentrations (95% CI: 5.9 to 20.0). Also, every increase of 1 BMI unit was associated with a decrease of 1.5 microgram/ml in the serum TCZ concentrations (95% CI: 0.8 to 2.2) and every increase of 1 kg in weight was associated with a decrease of 0.6 microgram/ml in the serum TCZ concentrations (95% CI: 0.3 to 0.9). Conclusions In the first year of TCZ treatment with a fixed dose regimen of 162 mg SC injection once a week, serum trough concentrations of TCZ are associated with CDAI scores and a state of CDAI remission/low disease activity. Body weight and BMI are inversely associated with serum TCZ concentrations. These results suggest that personalising the dose of SC TCZ to body weight may improve clinical outcomes of RA disease activity. Acknowledgements We thank Roche for the TASC study data sets. Disclosure of Interest None declared

Rheumatoid arthritis (RA) patients have many therapeutic options; however, tools to predict individual patient response are limited. The Genefron personal diagnostic kit, developed by analyzing large datasets, utilizes selected interferon stimulated gene expressions to predict treatment response. This study evaluates the kit's prediction accuracy of individual RA patients' response to tumor necrosis alpha (TNFα) blockers.A retrospective analysis was performed on RA patients reported in published datasets. A prospective analysis assessed RA patients, before and 3 months after starting a TNFα blocker. Clinical response was evaluated according to EULAR response criteria. Blood samples were obtained before starting treatment and were analyzed utilizing the kit which measures expression levels of selected genes by quantitative real time polymerase chain reaction (PCR). ROC analysis was applied to the published datasets and the prospective data.The Genefron kit analysis of retrospective data predicted the response to a TNFα blocker in 53 of 61 RA patients (86.8% accuracy). In the prospective analysis, the kit predicted the response in 16 of 18 patients (89% accuracy) achieving a EULAR moderate response, and in 15 of 18 patients achieving a EULAR good response (83.3% accuracy). ROC analysis applied to the two published datasets yielded an AUC of 0.89. ROC analysis applied to the prospective data yielded an AUC of 0.83 (sensitivity - 100%, specificity - 75%) The statistical power obtained in the prospective study was .9.The diagnostic kit predicted the response to TNFα blockers in a high percentage of patients assessed retrospectively or prospectively. This personal kit may guide selection of a suitable biological drug for the individual RA patient.

Galectin-3 is a β-galactoside-binding lectin that plays an important role in the modulation of immune responses. It has been shown to aggravate joint inflammation and destruction in experimental arthritis. We investigated the role of galectin-3 in TLR-induced cell activation in human synovial fibroblasts (SF) in order to better understand the mechanism(s) of the proinflammatory function of galectin-3 in arthritis. Galectin-3 expression in SF obtained from rheumatoid arthritis and osteoarthritis patients was inhibited by siRNA mediated gene-knockdown. Galectin-3 was also inhibited with modified citrus pectin (MCP), a polysaccharide galectin-3 ligand. Galectin-3 knockdown inhibited TLR-2, -3 and -4-induced IL-6 secretion, but not TLR-2, -3 and -4-mediated matrix metalloproteinase-3 or CC chemokine ligand-5 secretion. When the SF were stimulated with phorbol 12-myristate 13-acetate, a protein kinase C activator that bypasses the membranal receptors, galectin-3 knockdown no longer influenced IL-6 secretion. MCP reduced IL-6 levels in a dose-dependent manner. Our results indicate that galectin-3 is a positive sensor-regulator of TLR-induced IL-6 secretion in human synovial fibroblasts, thus adding new insights into the mechanisms by which galectin-3 augments synovial inflammation. These findings corroborate the potential role of glycan inhibitors of galectin-3 as a therapeutic approach for the treatment of inflammatory arthritis.

The objective of this study is to describe a series of patients above the age of 50 years with large vessel arteritis and vascular involvement typical of TAK. A retrospective review of 18 patients (median age 64 years) with emphasis on clinical characteristics, laboratory values, and vascular involvement by CT, MRI, or planar angiography. Five patients fulfilled the ACR criteria for GCA, five for TAK, three both GCA and TAK, while five patients did not fulfill the criteria for either disease. The dominant presenting symptoms were constitutional, while only a few patients had cranial or peripheral symptoms. Sixty-one percent had physical signs of vascular compromise. Temporal artery biopsy showed giant cell arteritis in six out of nine biopsies. Arterial involvement: 78 % had either involvement of the ascending aorta, the aortic arch, descending or/and abdominal aorta, 9 carotid, 12 subclavian, 5 axillary, 3 renal, 7 iliac, and 2 femoral arteries; 7 mesenteric or celiac trunk. All the patients were treated with prednisone and 50 % with steroid-sparing drug. Nine out of 15 patients (60 %) achieved remission after 1 year of follow-up. No substantial differences in the distribution of vascular involvement, type of treatment, or outcome measures were observed between patients fulfilling criteria for GCA or TAK. Vascular involvement typical of TAK in patients above the age of 50 years with large vessel arteritis seems to be more frequent than previously assumed. Our findings support the assumption that TAK and GCA represent a spectrum of the same disease.

Background Rheumatoid arthritis (RA) patients have many therapeutic options. However, there are limited tools to predict the individual patient9s response to therapy. The Genefron personal diagnostic kit (IFR 300) has been developed based on analysis of large databases to select interferon stimulated gene (ISG) expressions which could predict response to a biologic agent Objectives This study aims to evaluate the ability of the Genefron diagnostic kit to predict the individual RA patient response to TNFα blockers. Methods Two separate analyses were performed, one retrospective and one prospective analysis. The response of 61 RA patients reported in 2 published data sets was analyzed retrospectively utilizing the Genefron kit. In addition, 18 patients with RA were assessed prospectively, before and 3 months after starting treatment with a TNFα blocker. Clinical assessment included swollen and tender joint counts, patient and physician assessments of disease activity. Patients9 blood samples were obtained before administration of the TNFα blocker and were analyzed utilizing the Genefron diagnostic kit which measures expression levels of selected genes by quantitative real time PCR. Results Genefron kit analysis of retrospective data correctly predicted the response to a TNFα blocker in 53 of 61 RA patients (accuracy - 86.8%). In the prospective analysis 6 patients achieved a moderate EULAR response, 6 achieved a good EULAR response and 6 did not respond. According to the EULAR moderate response, the Genefron diagnostic kit predicted the response correctly in 16 of 18 patients (accuracy-89%, sensitivity -100%, specificity - 67%). According to the EULAR good response, the kit predicted the response correctly in 15 of 18 patients (accuracy - 83.3%, sensitivity - 100%, specificity - 75%). Conclusions The Genefron diagnostic kit predicted the response to TNFα blockers in a high percentage of RA patients assessed either retrospectively or prospectively in a real life setting. This personal diagnostic kit has the ability to guide selection of a suitable biological drug for the individual RA patient Disclosure of Interest None declared

The objective of the study was to combine ultrasonographic (US) with clinical findings for comparing the effect of adalimumab (ADA) to methotrexate (MTX) on the thickness of tendons and enthesis in psoriatic arthritis (PsA) patients. Forty-three consecutive PsA patients were examined at baseline and after 6 and 12 weeks of treatment with ADA or MTX. The US assessment included thickness measurement of the extensor (ET) and flexor tendons (FT) of the second and third finger of both hands, plantar aponeurosis (PA) and the Achilles tendon (AT) bilaterally. Disease activity (DA) was assessed by the number of tender (TJ) and swollen joints (SJ), the number of inflamed enthesis (IE), pain assessment (PAI), and patient (PDAI) and physician (PHDAI) disease activity evaluations by visual activity score (VAS). Nineteen patients received MTX and 24 patients received ADA. All DA parameters improved in both groups. A decrease in thickness of tendons and enthesis was observed only in the ADA group, reaching a level of significance for the left AT (p = 0.01), left PA (p = 0.007), the second left FT (p = 0.04) and the third ET (p = 0.04). ADA patients showed a trend towards a better response to treatment compared to MTX patients that reach significance at week 6 of treatment for the thickness of left AT (p = 0.04), left PA (p = 0.03), the number of TJ (p = 0.0136), PAI (p = 0.0028), and PDAI (p = 0.029). ADA treatment for PsA compared to MTX significantly improved signs of DA and several US parameters. US assessment of enthesis can be an additional useful tool in the monitoring of psoriatic enthesopathy.

Overproduction of extracellular diphosphate due to hydrolysis of ATP by NPP1 leads to pathological calcium diphosphate (pyrophosphate) dihydrate deposition (CPPD) in cartilage, resulting in a degenerative joint disease that today lacks a cure. Here, we targeted the identification of novel NPP1 inhibitors as potential therapeutic agents for CPPD deposition disease. Specifically, we synthesized novel analogs of AMP (NPP1 reaction product) and ADP (NPP1 inhibitor). These derivatives incorporate several chemical modifications of the natural nucleotides including (1) a methylene group replacing the Pα,β-bridging oxygen atom to provide metabolic resistance, (2) sulfonate group(s) replacing phosphonate(s) to improve binding to NPP1’s catalytic zinc ions, (3) an acyclic nucleotide analog to allow flexible binding in the NPP1 catalytic site, and (4) a benzimidazole base replacing adenine. Among the investigated compounds, adenine-N9-(methoxy)ethyl-β-bisphosphonate, 10, was identified as an NPP1 inhibitor (Ki 16.3 μM vs. the artificial substrate p-nitrophenyl thymidine-5′-monophosphate (p-Nph-5′-TMP), and 9.60 μM vs. the natural substrate, ATP). Compound 10 was selective for NPP1 vs. human NPP3, human CD39, and tissue non-specific alkaline phosphatase (TNAP), but also inhibited human CD73 (Ki 12.6 μM). Thus, 10 is a dual NPP1/CD73 inhibitor, which could not only be of interest for treating CPPD deposition disease and calcific aortic valve disease but may also be considered for the immunotherapy of cancer. Compound 10 proved to be a promising inhibitor, which almost completely reduces NPPase activity in human osteoarthritic chondrocytes at a concentration of 100 μM.

Dysregulated activity of A Disintegrin And Metalloproteinase 17 (ADAM17)/TNFα Converting Enzyme (TACE) is associated with inflammatory disorders and cancer progression by releasing regulatory membrane-tethered proteins like TNFα, IL6R and EGFR ligands. Although specific inhibition of TACE is thought to be a viable strategy for inflammatory disorders and for malignancies treatment, the generation of effective inhibitors in vivo has been proven to be challenging. Here we report on the development of a protein inhibitor that leverages the endogenous modulator of TACE. We have generated a stable form of the auto-inhibitory TACE prodomain (TPD), which specifically inhibits in vitro and cell-surface TACE, but not the related ADAM10, and effectively modulated TNFα secretion in cells. TPD significantly attenuated TACE-mediated disease models of sepsis, rheumatoid arthritis (RA) and inflammatory bowel disease (IBD), and reduced TNFα in synovial fluids from RA patients. Our results demonstrate that intervening with endogenous ADAM sheddase modulatory mechanisms holds potential as a general strategy for the design of ADAM inhibitors.

Objective.To estimate the longterm humoral response of an antipneumococcal polysaccharide vaccine (PPSV23) in patients with rheumatoid arthritis (RA), psoriatic arthritis (PsA), ankylosing spondylitis (AS), or inflammatory bowel disease (IBD)-associated spondyloarthropathy (SpA), and the effect of demographic and clinical factors and treatment on the longterm efficacy of the vaccine.Methods.A total of 145 consecutive patients treated with biologics [tumor necrosis factor-α (TNF-α) or interleukin 6 (IL-6) receptor inhibitors] or methotrexate (MTX) participated in this study. Fifteen were excluded because of absent information regarding their vaccination status (n = 9) or because of technical problems in obtaining their serum sample (n = 6). They were diagnosed with RA (n = 63, 48.5%), PsA (n = 29, 22.3%), AS (n = 28, 21.5%), or IBD-associated SpA (n = 3, 2.3%). Their mean age was 54.6 years, and 61.5% were women. Data were collected on the timing of vaccination, demographic and clinical characteristics, and treatment, and patients’ serum antipneumococcal antibody levels were tested.Results.Two-thirds of the patients (67.7%) had received PPSV23 45 months (mean) earlier. Treatment included TNF-α inhibitors (73.9%), IL-6 receptor inhibitors (13.1%), or MTX without a biological treatment (13%). The uptake of vaccination was significantly higher in the older population (> 65 yrs). Vaccinated patients had significantly higher antibody levels compared with vaccine-naive patients. The antibody levels had been preserved after 10 years. MTX use, but not biologics, was associated with significantly lower antibody levels.Conclusion.The longterm efficacy of the PPSV23 vaccination seems to be preserved among patients with RA, PsA, AS, and IBD-associated SpA for at least 10 years. Efficacy is slightly impaired by MTX, but it is not affected by biologics. These findings suggest that revaccination after 5 years might not be needed for all, and testing the antibody titers should be considered to identify those who may benefit from revaccination.

Objective: Monitoring of immune responses is essential in the care of immunosuppressed individuals, including rheumatic patients. Evaluation of cellular immunity is essential for confirming virus-specific effector cell functions, but it is poorly standardized, and suffers from technical limitations and inaccurate results. There is, therefore, a need for reliable techniques for assessing cell-mediated immunity. In this study we compared the cell-mediated immunity response to influenza vaccine between a population of rheumatoid arthritis (RA) patients and healthy subjects by three methods. Methods: Trivalent influenza subunit vaccine was administered to 18 RA patients who were taking disease-modifying antirheumatic drugs and to 18 healthy controls. Peripheral blood mononuclear cells (PBMCs) and sera were obtained immediately before and ~28 days after vaccination. Cell-mediated immunity responses to vaccination were evaluated by (1) flow cytometric analysis of IL-2/IFN-γ production in activated CD4/CD8 T-cells, (2) enzyme-linked immunosorbent assay for the analysis of IFN-γ secretion, and (3) Granzyme B activity assay. Humoral response was evaluated by the hemagglutination inhibition assay. Results: Vaccination induced a significant increase in PBMC IFN-γ secretion and Granzyme B activity in the RA patients. Granzyme B activity also significantly increased in the controls, but there was no change in the levels of secreted IFN-γ. No group differences in the frequencies of IFN-γ/IL-2-producing activated CD4/CD8 T-cells were observed by flow cytometry. The geometric mean of hemagglutination inhibition antibody titers increased significantly for the H1N1/H3N2 influenza strains in both groups. Conclusions: Granzyme B activity assay was the only method to detect a significant cell-mediated immunity response in both groups while significant increase in IFN-γ secretion was demonstrated only in RA patients. Flow cytometric analysis failed to show IL-2 and IFN-γ production in both groups. Currently available methods for measuring cellular responsiveness to influenza vaccination are inconsistent and limited in their ability to reflect acquired cellular immunity.

Objective.The longterm use of tumor necrosis factor (TNF)-α blockers is limited by the formation of neutralizing antibodies. To the best of our knowledge, immunogenicity in psoriatic arthritis (PsA) has not been investigated in depth. Our objective was to evaluate the prevalence and significance of TNF-α blocker immunogenicity in PsA.Methods.Consecutive patients with PsA treated with either infliximab (IFX), adalimumab (ADA), or etanercept (ETN) > 3 months participated in our cross-sectional study. Their demographic and clinical characteristics, skin and joint disease activity, and records of use of methotrexate (MTX) and other medications were collected. Drug levels (ELISA) and antidrug antibodies (ADAb; Bridging ELISA) were evaluated before the next injection or infusion.Results.A total of 93 patients with PsA were recruited (48 receiving ADA, 24 IFX, and 21 ETN), with a mean age of 53 years (range 21–83 yrs), composed of 53% women. One-fourth of the patients were concomitantly treated with MTX. Altogether, 77% of the patients demonstrated therapeutic drug levels. High levels of ADAb were found in 29% of patients taking ADA, 21% taking IFX, and 0% taking ETN. ADAb significantly correlated with lower drug levels, higher 28-joint Disease Activity Scores, and higher global assessments. MTX use correlated significantly with a lower prevalence of ADAb.Conclusion.Significant levels of ADAb were present in up to 29% of patients with PsA treated with ADA or IFX. ADAb clearly correlated with low therapeutic drug levels and higher disease activity variables. The use of MTX significantly decreased ADAb prevalence, and its use should be strongly considered in combination with TNF-α blocker antibodies in patients with PsA.

We aimed to assess the immunogenicity and safety of vaccination against seasonal influenza in psoriatic arthritis (PsA) and psoriasis (Pso) patients.Patients with PsA or Pso and healthy controls were vaccinated with the Sanofi Pasteur vaccine recommended by the WHO in 2012. Clinical and laboratory assessments were performed on the day of the vaccination and 4-6 weeks later. The immunogenicity of the vaccine was evaluated by haemagglutination inhibition assay.The study included 63 consecutive PsA patients and 4 Pso patients (mean age 50.1, 37 females, 30 males, 55.2% treated with tumour necrosis factor alpha blockers [TNF-α], 31.3% on disease-modifying anti-rheumatic drugs [DMARDs]) and 30 healthy controls. The geometric mean titers increased significantly in all participants for each of the subtypes tested. A substantial and similar proportion of patients in both groups responded to the vaccine. The response rate was not affected by parameters such as age, gender, disease activity or the use of TNF-α blockers or DMARDs. There were no significant changes in the patients' 68 tender and 66 swollen joint counts, dactylitis, PASI, global evaluation of the patient and physician and ESR, while there was a rise in CRP levels.Vaccination against seasonal influenza is safe and induces an appropriate response in patients with PsA, similar to healthy controls.

We present the technological merits of combining a high fill factor, one-dimensional, MEMS micro-mirror array for switching and liquid crystal shutters for amplitude control within a wavelength-selective switch (WSS). The combination allows for simplified MEMS actuator design, support for high port counts, high attenuation accuracy, high natural frequency, mechanical robustness against shock & vibrations, and fast switching times. The complexity associated with dual modulating elements at the focal plane is addressed by design choices and optimization of the depth of focus. The WSS is fully Telcordia qualified and carrying live traffic in deployed networks.

Background Osteoarthritis (OA) affects hundreds of millions of people worldwide, resulting in significant morbidity. Articular cartilage homeostasis is governed by resident chondrocytes that can either contribute to cartilage anabolism by producing tissue constituents or drive catabolic events by secreting aggrecanases and collagenases. The chondrocytes9 pericellular matrix (PCM) acts as a mechanosensor by sequestering growth factors that are bound to heparan sulfate (HS) proteoglycans, such as perlecan. Heparanase is the sole mammalian enzyme with HS degrading activities. Heparanase enzymatic activity releases HS-bound growth factors, cytokines, chemokines and other ligands stored within the extracellular matrix. Through both enzymatic and non-enzymatic activities, heparanase influences the expression of a number of important genes and has been implicated in numerous processes involved in cancer progression, inflammation and remodeling of the ECM. Little is known about whether heparanase plays a role in cartilage homeostasis. Objectives To ascertain whether heparanase may have a role in modulating the anabolic or catabolic responses of human articular chondrocytes. Methods Heparanase enzymatic activity was measured in collagenase digests of osteoarthritic articular cartilage using solid-phase-bound radiolabeled heparan sulfate as substrate. Primary human chondrocytes obtained from OA patients undergoing joint replacement surgery, were incubated with or without recombinant human pro-heparanase. In some experiments, the heparanase inhibitor, PG545, was also added. Heparanase was also added together with fibroblast growth factor 2 (FGF-2) or interleukin 1 (IL-1). The association of pro-heparanase with the chondrocytes and its conversion to active heparanase was evaluated by Western blot. Induction of the catabolic genes MMP13 , ADAMTS4 and ADAMTS5 and the anabolic genes ACAN (encoding the aggrecan core protein) and COL2A1 (encoding the alpha chain of type II collagen), was analyzed by real time reverse-transcriptase PCR. MMP13 enzymatic activity in the culture medium was measured with a specific fluorescent assay. ERK phosphorylation was evaluated by Western blot. Results The collagenase-digested cartilage (but not the collagenase blend alone), liberated small fragments compatible with HS-degradation products, indicated active heparanase is present in osteoarthritis cartilage. Cultured chondrocytes rapidly associated with the inactive 62-kDa pro-heparanase, which was then cleaved to the active 50-kDa enzyme. Addition of heparanase to cultured human chondrocytes activated the catabolic genes MMP13 and ADAMTS4 by 4 hours and reduced the expression of anabolic genes ACAN and COL2A1 by 20 hours. Heparanase also induced human chondrocytes to secrete active MMP13 to the culture medium by 20 hours and induced intracellular ERK phosphorylation by 30 minutes. A synergistic effect was observed between heparanase and FGF-2, but not between heparanase and IL-1. PG545, a heparan sulfate mimetic, reversed the effects of heparanase. Conclusions Heparanase is active in osteoarthritic cartilage and induces catabolic responses in cultured primary human chondrocytes. This response seems to be due, at least in part, to the release of heparan sulfate-bound growth factors such as FGF-2. Disclosure of Interest None declared

Monogenic periodic fever syndromes are characterized by recurrent episodes of fever, accompanied by localized inflammatory manifestations. Among them, familial Mediterranean fever (FMF) is the most studied and is by far the most prevalent periodic fever syndrome in Israel. We present a diagnostic workup of a patient suffering from a periodic fever syndrome, initially thought to be FMF and characterized by attacks of fever, severe abdominal pain, a migratory erythematous rash and conjunctivitis. The development of periorbital edema presenting as diplopia led to consideration of tumor necrosis factor receptor-1-associated periodic syndrome (TRAPS). Genetic tests confirmed the diagnosis. This case should alert us that even in Israel, a patient with periodic fever not fully consistent with the typical features of FMF, should be evaluated for other periodic fever syndromes.

Objective To assess the efficacy and safety of vaccination against pandemic H1N1 virus in patients with rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), psoriatic arthritis (PsA), and ankylosing spondylitis (AS) compared with healthy controls. Methods The study population comprised 41 RA patients, 21 SLE patients, 17 PsA patients, 15 AS patients, and 25 healthy controls. All were vaccinated using the Novartis MF59-adjuvanted H1N1v monovalent influenza vaccine. The immunogenicity of the vaccine was assessed on day 1 and again 4 weeks later by hemagglutination inhibition assay. Geometric mean titers and seroconversion rates were calculated for each group. The safety of the vaccine was evaluated using the 28-joint Disease Activity Score (DAS28) for RA and PsA, the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI), and the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI). Results The proportion of baseline protective levels of antibodies against H1N1 was similar in all but the AS group, in which it was lower. The geometric mean titers increased significantly in all 5 groups. A substantial proportion of patients and controls responded to the vaccine. The healthy controls demonstrated a better response than each of the other groups: 84% versus 56% for RA, 67% for SLE, 59% for PsA, and 53% for AS. Multivariate logistic regression analysis identified RA and PsA as parameters of significantly lower response. The DAS28, BASDAI, and SLEDAI remained unchanged after vaccination. Conclusion Vaccination against pandemic H1N1 using an adjuvanted H1N1v monovalent influenza is safe and induced an appropriate response in patients with RA, SLE, PsA, and AS.

Objectives Yearly vaccination against influenza is currently recommended to patients with rheumatoid Arthritis (RA). Antibody and cell-mediated responses are both involved in the defense against influenza. Humoral responses to influenza vaccine are impaired in RA patients treated with rituximab (RTX). The objectives of this study were to comparatively assess cell mediated and humoral responses to influenza vaccination in RA patients with or without RTX-induced CD20 B-cell depletion. Methods Trivalent influenza subunit vaccine was administered to 46 RA patients and to 16 healthy controls. The RA group included 29 patients treated by RTX and 17 on conventional disease-modifying anti-rheumatic drugs (DMARDs), mostly methotrexate. Peripheral blood mononuclear cells and sera were obtained immediately before and 4–6 weeks after vaccination. Cell-mediated response to influenza antigens was evaluated by flow cytometry for activated CD4 T-cells. Humoral response was evaluated by haemagglutination inhibition assay. Results Cellular response : Cell-mediated responses were comparable in RTX-treated vs. DMARDs-treated patients. The recall postvaccination CD4+ cellular response was similar in RA patients and healthy controls. A positive correlation was found between CD19+ cell count on the day of vaccination and cellular response in RTX-treated RA patients. Humoral response : The antibody response rate was significantly impaired in the RTX group: being 26.4%, 68.4% and 47.1% in RTX-treated, DMARDs-treated and controls, respectively. Conclusion Cellular immunity to influenza vaccination in RTX-treated patients was similar to DMARDs-treated patients and healthy controls, while humoral immunity was severely impaired. The preservation of cellular immunity may explain the relatively low rate of infection among B-cell depleted patients.

Objectives To report 3 cases ANCA-associated vasculitis (AAV) that developed in patients suffering from systemic sclerosis (SSc) and to review previously reported cases. Methods We describe 3 patients diagnosed with SSc who developed severe AAV that presented as crescentic glomerulonephritis (GN) and/or alveolar hemorrhage. A retrospective review of the literature was performed using the PubMed database. Results The first patient presented with rapidly progressive renal failure and then with 2 episodes of massive alveolar hemorrhage. She was partially refractory to treatment with corticosteroids and cyclophosphamide but responded promptly to treatment with rituximab. The second patient suffered from 2 episodes of fulminant alveolar hemorrhage; the first responded to intravenous corticosteroids, but the second was fatal despite aggressive immune suppression with corticosteroids and cyclophosphamide. The third patient presented with a clinical picture compatible with scleroderma renal crisis (SRC) but was later diagnosed with crescentic GN and subsequently died from probable alveolar hemorrhage. Thirty-seven cases of AAV in SSc patients have been described in the English literature. Clinical manifestations include rapidly progressive GN, alveolar hemorrhage, limb ischemia, and vasculitic skin rash. In contrast to SRC that usually develops early in the course of SSc, ANCA-associated GN in SSc patients occurred later, after several years of illness. Hypertension, microangiopathic hemolytic anemia, and thrombocytopenia that are the hallmark of SRC were observed only in a minority of AAV cases. Almost all cases of AAV in SSc were positive for MPO-ANCA. Conclusions AAV in the setting of SSc is a diagnostic challenge. Differential diagnosis from SRC is crucial as treatment approach for these conditions completely differs. High doses of corticosteroids and immune suppression are advocated in severe AAV. In resistant cases, treatment with rituximab may be considered.

Background Calcium pyrophosphate deposition (CPPD) is associated with osteoarthritis (OA) and is the cause of a common inflammatory articular disease (1). Nucleotide pyrophosphatase/phosphodiesterase1 (eNPP1) is the major ecto-pyrophosphatase (NTPPPH) in chondrocytes and chondrocyte-derived matrix vesicles (MVs) (2). Thus, eNPP1 is a principle contributor to extracellular pyrophosphate levels and a potential target for interventions aimed at preventing CPPD. Recently, we synthesized and described a novel eNPP1 inhibitor (3). Objectives To evaluate whether the eNPP1-specific inhibitor, SK4A, attenuates NTPPPH activity in human OA articular cartilage. Methods Cartilage tissue, primary chondrocytes and cartilage-derived MVs were obtained from donors with osteoarthritis undergoing knee replacement surgery.The effect of SK4A on cell viability was assayed by the XTT method. eNPP1 expression was evaluated by Western blot. NTPPPH activity was measured by a colorimetric assay with thymidine 59-monophosphate p-nitrophenyl ester as a substrate and by HPLC analysis of ATP levels. Results Primary OA chondrocytes express eNPP1 in early passages, but this expression is subsequently lost upon further passaging. Similarly, significant NTPPPH activity can only be detected in early-passage human chondrocytes. The eNPP1 inhibitor, SK4A, does not affect chondrocyte viability at concentrations of up to 1mM. SK4A effectively inhibits the ex vivo NTPPPH activity in whole cartilage tissue, in isolated primary chondrocytes and in cartilage-derived MVs. Conclusions SK4A inhibits NTTTPH activity in human OA cartilage and may prevent extracellular pyrophosphate accumulation and calcium pyrophosphate deposition. References Zhang W, Doherty M, Bardin T, et al. Ann Rheum Dis. 2011;70(4):563-70. Johnson K, Terkeltaub R. Front Biosci. 2005; 10:988-97. Nadel Y, Lecka J, Gilad Y, et al. J Med Chem. 2014; 57(11): 4677-91. Disclosure of Interest None declared

Efficient reconstitution of defective genes in hepatocytes could be used to treat various liver and systemic diseases through gene therapy. To explore the potential of SV40-based vectors in liver gene therapy, we constructed SV/luc, an SV40 T-antigen replacement transduction vector, that was propagated on COS and COT cells, which supply the SV40 T-antigen in trans. For liver targeting, BALB/C mice were injected via the tail vein with SV/luc stocks containing 3 x 10(6) to 10(8) transducing units in a volume of 1-2 ml. Luciferase activity was monitored with a light-detection cooled charged-coupled device (CCCD) camera, which enables continuous in vivo measurement of luc expression. The SV40 vector proved to be efficient in gene delivery to the liver, leading to long-term (or =107 days) transgene expression in hepatocytes. Optimal results were obtained with 3 x 10(6) to 3 x 10(7) transducing units. The hydrodynamic vector delivery method caused transient liver inflammatory changes, with full recovery within days. Low levels of SV40-neutralizing antibodies were detected in the sera of treated mice; however, there was no indication of vector or transgene-specific cellular immune responses. Vectors packaged in vitro, using recombinant capsid proteins and plasmid DNA, were also effective in liver transduction. These results suggest that SV40 vectors may be useful for liver gene therapy.

Background/Aims : Chronic HBV infection, a world-wide epidemic, can lead to chronic hepatitis and eventually to cirrhosis and hepatocellular carcinoma. The liver poses obstacles for many available gene-transfer vectors. SV40-based vectors can transduce human hepatic and hematopoietic cells. We studied the effect of HBV on the transduction – efficiency of human hepatic cells by SV40 – based vectors. Methods : A SV40-vector carrying the luciferase gene, and wild-type SV40, were used to assess transduction efficiency of human HBV-positive and HBV-negative hepatic cells. Transduction efficiency was measured as luciferase activity or by T-antigen staining. To evaluate whether differences in transduction efficiency are due to cell recognition and/or nuclear transport, MHC-I receptors were measured by FACS analysis and SV40-DNA was extracted from the nuclei of transduced cells and quantified. Results : Two HBV-positive cell-lines, HepG2.2.2.15 and FLC4-A10II, were transduced significantly more efficiently than their parental HBV-negative cell-lines. Transient transfection of HuH-7 cells with the HBV genome also increased transduction efficiency. The level of MHC-I, the cellular receptor for SV40, was comparable in all the cell-lines studied. However, soon after infection with SV40, the nuclei of HepG2.2.2.15 contained >6-fold more SV40-DNA than HepG2. Conclusions : HBV increases transduction by SV40-vectors. This is due to enhanced vector entry and/or transport into the nucleus. SV40-vectors appear to have a potential for gene therapy for the treatment of HBV infections.

Simian virus 40 (SV40) vectors are efficient vehicles for gene delivery to hematopoietic and hepatic cells. To ensure their replication incompetence and because of safety considerations, it is critical that the vectors do not contain T-antigen sequences. Available packaging cell lines for T-antigen replacement vectors, COS and CMT4, contain considerable sequence identity with the vectors, leading to homologous recombination and reacquisition of the T-antigen gene. We constructed a packaging cell line, COT18, with minimal sequence identity to the vector. Vector stocks produced by passaging on COT18 had high transducing activity and undetectable levels of T-antigen-positive, replication-competent contaminants. This cell line provides a means for the preparation of safe SV40 vector stocks.

The problem of finding a large independent set in a hyper-graph by an online algorithm is considered. We provide bounds for the best possible performance ratio of deterministic vs. randomized and non-preemptive vs. preemptive algorithms. Applying these results we prove bounds for the performance of online algorithms for routing problems via fixed paths over networks.

Background Galectin-3 is a β-galactoside-binding lectin that plays an important role in the modulation of immune responses. Galectin-3 levels are increased in rheumatoid arthritis (RA) synovial tissue, synovial fluid and peripheral blood 1 . Recombinant exogenous galectin-3 stimulates pro-inflammatory cytokine secretion by fibroblast-like synoviocytes (FLS) 2 . In a murine, antigen-induced arthritis model, Gal-3-/- mice displayed significantly reduced synovitis and joint erosion compared to WT mice 3 . Objectives To examine the effect of galectin-3 inhibition on toll-like receptor (TLR)-induced secretion of interleukin-6 (IL-6) in FLS from RA and osteoarthritis (OA) patients. Methods FLS from RA and OA patients were harvested from synovial fluid aspirates or directly from synovial tissue obtained during orthopedic surgery. Galectin-3 was inhibited either by siRNA mediated gene-knockdown, or by the galectin ligands thiodigalactoside and modified citrus pectin (MCP). The cells were then stimulated with ligands for TLR-2, 3 and 4; peptidoglycan (PGN), poly(I:C) and lipopolysaccharide (LPS), respectively, and the levels of IL-6 secretion were measured by ELISA. Results Galectin-3 gene knockdown inhibited TLR-2, 3 and 4 induced IL-6 secretion in FLS. Treatment with thiodigalactoside, a galectin oligosaccharide ligand, also decreased TLR-meditated IL-6 secretion. Treatment with MCP, a polysaccharide galectin-3 ligand, attenuated IL-6 secretion due to stimulation with PGN and poly(I:C), but augmented IL-6 secretion following LPS-stimulation, in a dose dependent manner. Conclusions Galectin-3 modulates TLR-induced IL-6 secretion in FLS. Knockdown of galectin-3 inhibits PGN, poly(I:C) and LPS-induced IL-6 secretion. Galectin-3 saccharide ligands can also inhibit TLR-induced IL-6 secretion. Understanding the role of galectin-3 in TLR-induced cytokine secretion may add new insights into the pathological mechanisms of autoimmunity and perhaps aid in the design of galectin-3 inhibitors as potential therapeutics. References Ohshima S, Kuchen S, Seemayer CA, et al. Galectin 3 and its binding protein in rheumatoid arthritis. Arthritis Rheum. 2003; 48(10): 2788-95. Filer A, Bik M, Parsonage GN, et al. Galectin 3 induces a distinctive pattern of cytokine and chemokine production in rheumatoid synovial fibroblasts via selective signaling pathways. Arthritis Rheum. 2009; 60(6): 1604-14. Forsman H, Islander U, Andreasson E, et al. Galectin 3 aggravates joint inflammation and destruction in antigen-induced arthritis. Arthritis Rheum. 2011; 63(2): 445-54. Disclosure of Interest None Declared

Background Takayasu arteritis (TAK) and Giant cell arteritis (GCA) are classified as two different vasculitides.[1] The differentiation is based on age at onset (under 40y for TAK, above 50y for GCA) and vascular distribution (aorta and its primary branches for TAK and mainly extracranial arteries for GCA). However, both diseases involve large vessels and share histopathology findings. Objectives Our goal is to describe a series of patients above the age of 50y with large vessel arteritis and vascular involvement typical of TAK. Methods Eighteen Patients (median age 64y, 16 females) were selected from 3 centers in Israel. A retrospective review of the cases was preformed with special emphasis on clinical characteristics (including clinical course and outcome), laboratory values and vascular involvement. Vascular imaging by CT angiography, MRI angiography or planar angiography was done in all cases. Results Five patients fulfilled the ACR criteria for GCA, 5 the ACR criteria for TAK, 3 both GCA and TAK criteria, while 5 patients had clear involvement of large vessels without fulfilling the criteria for either disease. The dominant presenting symptoms were constitutional, while only a few patients had cranial or peripheral symptoms. On physical examination, 61% had signs of vascular compromise including: blood pressure arm differences, non palpable limb pulses or vascular bruits. All patients had elevated acute phase reactants, 78% had anemia. Temporal artery biopsy showed giant cell arteritis in 6 out of 9 biopsies. The majority (78%) showed either involvement of the ascending aorta, the aortic arch, descending aorta or a combination of these. The distribution of vessel involvement was as following: carotid arteries in 9 patients, subclavian arteries in 12, axillary arteries in 5, abdominal aorta in 9, mesenteric or celiac trunk in 7, renal arteries in 3, iliac arteries in 7 and femoral arteries in 2. All the patients were treated with prednisone and 50% with steroid sparing drug. Nine out of 15 patients (60%) that have completed at least 1 year of follow-up achieved remission. Three additional patients had a partial remission. Three patients of the remission group relapsed. A comparison between the patients who fulfilled the ACR criteria for GCA and those who fulfilled for TAK showed that the first group was slightly older (70y vs. 64y) and had more constitutional symptoms while the second group had more peripheral symptoms and more vascular involvement signs in physical examination. The acute phase reactants were slightly higher in the GCA group. However, no substantial differences in the distribution of vascular involvement, type of treatment or outcome measures were observed. Conclusions Vascular involvement typical of TAK in patients above the age of 50y with large vessel arteritis seems to be more frequent than previously assumed. The differences between those who fulfill the criteria for each disease are minor. Our findings support the assumption that TAK and GCA represent a spectrum of the same disease. References Fries JF, Hunder GG, Bloch DA, et al. The ACR 1990 criteria for the classification of vasculitis. Arthritis Rheum. 1990;33:1135-6. Disclosure of Interest None Declared

Background Screening for latent tuberculosis (TB) is mandatory before treatment with TNFα blockers. It has been suggested that latent TB based on tuberculin skin test (TST) may be over diagnosed due to Koebner phenomenon in patients with psoriasis. Objectives To assess the effect of the extent of skin and joint involvement on the diagnosis of latent TB and to compare the TST and QuantiFERON-TB Gold (QTF-G) in patients with psoriasis (PSO) and psoriatic arthritis (PsA) to rheumatoid arthritis (RA) patients. Methods Forty-one PSO patients (35 with PsA) and 35 RA patients candidates to receive TNF α were prospectively evaluated for TST, QTF-G, risk factors for latent TB as well as for the severity of the underlying diseases using psoriasis activity severity index (PASI) and the number of painful and swollen joints. Results 41 PSO patients (mean age 49.7, 48% women, 36% treated with MTX) were compared to 35 RA patients (mean age 60 (p=0.006), 70% women, 70% treated with MTX (p=0.02)). No differences were observed in the mean TST (5±6.7 for PSO, 5.8±6.3 for RA), the proportion of positive TST based on 5mm as well as 10 mm, the proportion of QTF-G positive patients (7.2% in PSO vs 12% in RA). The level of agreement between TST and QTF-G was similar between the 2 groups (using TST 5 and TST 10, 74% and 81% for PSO vs 60% and 76% for RA, respectively). Furthermore, no correlation was found between the extent of psoriasis involvement evaluated by PASI and the TST, QTF-G, and the level of agreement between them. The number of tender and swollen joints, the use of MTX or corticosteroids did not affect the diagnosis of screening TB Conclusions The extent of severity of psoriasis does not affect the results of TST and QTF-G in patients with psoriasis. The level of agreement between TST and QTF-G was similar in RA and PSO. Although it remains to be determined with test is preferable in the diagnosis of latent TB, whether the patient suffers from PSO/PsA or RA does not seem to play an important role in this choice. Disclosure of Interest None Declared

Background Monitoring of immune responses is essential in a number of clinical research areas that address immunosuppressed individuals, including oncologic and rheumatic patients. During the past 10 years, our group has been deeply involved in the study of vaccinations in rheumatic patients, with a special concern regarding the impact of drugs. While humoral responsiveness is considered the gold standard for most vaccines, including seasonal influenza, evaluation of cellular immune responses is much less standardized, requires specialized equipment and often suffers from technical limitations that eventually lead to inaccurate and inconsistent results. Since cellular response is essential for supporting virus-specific effector cell functions, reliable assessment techniques are of great importance. Objectives To comparatively assess the cellular immune response to influenza vaccine by 3 different methods, in concomitant with humoral immune response evaluation, in rheumatoid arthritis (RA) patients relative to healthy subjects. Methods Trivalent influenza subunit vaccine was administered to 18 RA patients treated by synthetic DMARDs, mostly methotrexate (MTX), and to 18 healthy controls. Peripheral blood mononuclear cells (PBMCs) and sera were obtained immediately before and ~28 days after vaccination. Three different methods to measure cellular mediated immune responses to influenza vaccination were compared in RA patients and in healthy subjects. These methods included (1) flow cytometric analysis of IL-2 and IFNg production in activated CD4/CD8 T-cells; (2) enzyme-linked immunosorbent assay (ELISA) for the analysis of IFNg secretion and (3) granzyme B activity assay in influenza-stimulated PBMC. The humoral response was evaluated by the hemagglutination inhibition (HI) assay. Results Cellular response: Vaccination induced a significant increase in PBMC IFNg secretion and Granzyme B activity in the RA patients. A significant increase in Granzyme B activity was also measured in the healthy control group, but there was no change in the levels of secreted IFNg. There was no difference in the frequencies of IFN-g/IL-2 producing activated CD4 or CD8 T-cells, as measured by flow cytometry, in both groups, either before or 4 weeks following influenza vaccination. Humoral response: A significant increase in the geometric mean titer (GMT) of HI was demonstrated for the H1N1 and H3N2 influenza strains in both groups. The response rate to HIN1 was significantly higher in DMARDs patients in comparison with healthy controls. The response rate to the H3N2 and B strains was similar in both groups. Conclusions The results of this study confirm previous findings on the preserved humoral immune response of RA patients to influenza vaccination. Among the three methods utilized to cellular immune responsiveness - Granzyme B activity assay was the only one to detect such response in both groups tested. It appears that current available measurement methods for cellular responsiveness to influenza vaccination suffer from inconsistency, and are limited in their ability to reflect gained cellular immunity. Disclosure of Interest None Declared

Background Diffuse idiopathic skeletal hyperostosis (DISH) is a non-inflammatory condition that involves calcification and ossification of the spinal ligaments and entheses. In axial spondyloarthritis (SpA), an inflammatory disease associated with spinal ankylosis and enthesopathies, the pattern of new bone formation seen in the spine is different. While, characteristic magnetic resonance imaging (MRI) lesions of the spine in patients with SpA have been extensively described and defined 1 , there is a paucity of information regarding the MRI findings in DISH. Objectives To describe the MRI findings of patients with a radiographic diagnosis of DISH. Methods We collected computed tomography studies with findings characteristic of DISH (according to the classification criteria set by Resnick and Niwayama 2 ) and that also had corresponding and concurrent MRI studies of the spine. For each patient, sagittal T1 and STIR MRI sequences were evaluated for anterior/posterior spondylitis and marrow fat depositions at the vertebral corners. Results Imaging studies of 10 patients had both radiographic evidence of DISH and available MRI studies of the spine. The male:female ratio was 4:1 and the median age was 70 years (range 64-82). The indications for imaging included back pain (n=4), trauma (n=2), suspected space occupying lesion (n=2), osteomyelitis (n=1) and atelectasis (n=1). Vertebral corner fat depositions were detected in 8 patients. In 3 of these patients, at least 3 vertebrae were involved, a finding suggestive of axial SpA according to the ASAS/OMERACT consensus statement 2 . Lesions consistent with anterior/posterior spondylitis at vertebral corners were detected in 3 patients, and in one patient lesions were detected in 3 separate vertebrae. Conclusions In a cohort of patients with DISH, about a third display spinal MRI lesions that are characteristic of axial spondyloarthritis. This bears relevance to cases with diagnostic uncertainty and may imply overlapping pathogenetic mechanisms for new bone formation in both SpA and DISH. Further study is indicated to better characterize the similarities and differences between the MRI lesions of DISH and SpA. References Hermann KG, Baraliakos X, van der Heijde DM, et al. Descriptions of spinal MRI lesions and definition of a positive MRI of the spine in axial spondyloarthritis: a consensual approach by the ASAS/OMERACT MRI study group. Ann Rheum Dis. 2012;71:1278-88. Resnick D, Niwayama G. Radiographic and pathologic features of spinal involvement in diffuse idiopathic skeletal hyperostosis (DISH). Radiology. 1976;119:559-568. Disclosure of Interest None Declared