Renata Sokolik

The Activity of JAK/STAT and NF-κB in Patients with Rheumatoid Arthritis.

BACKGROUND Research is still being conducted in order to determine the mechanisms responsible for the initiation of rheumatoid arthritis (RA) as well as for its persistence and progression. OBJECTIVES The aim of this work was to establish the expression of the signal transducer and activator of transcription (STAT) transcription factors and the nuclear factor kappa-light-chain-enhancer of activated B cells (NFκB) transcription factor in peripheral blood leukocytes and synovial fluid cells. The correlations between the activation level of the transcription factors and the activity of the disease were also analyzed. MATERIAL AND METHODS In total, the study included 34 RA patients and 19 healthy individuals as controls. The expression of NFκB, STAT1, STAT3, STAT4, STAT5 and STAT6 in peripheral blood leukocytes and synovial fluid cells was established. The immunocytochemistry method was used to determine the degree of activation of STAT and NF-κB transcription factors. For the location of the factors, primary polyclonal anti-STATs and monoclonal anti-NF-κB antibodies were used. RESULTS The expression of STAT1, STAT3, STAT4, STAT5, STAT6 and NFκB was significantly higher in the group of RA patients than in the controls. No statistically significant differences were found between the expression of STATs in peripheral blood leukocytes and synovial fluid cells. CONCLUSIONS In comparison with the control group, the expression of the STAT and NFκB transcription factors in RA patients was higher, which may be helpful in better understanding the etiopathogenesis of the disease in the future, and may potentially have important therapeutic implications.

Significance of association of HLA-C and HLA-E with psoriatic arthritis

Psoriatic arthritis (PsA) is a complex genetic disorder that results from an interplay between multiple genetic and environmental factors. The aim of the study was to assess the significance of the association between the HLA-C and HLA-E allelic groups and PsA. Our results confirm the association between HLA-C(∗)06 and PsA (OR=5.16, p<0.0001). Furthermore, HLA-C(∗)06-positive patients develop more severe disease (p<0.01) and more frequently present with polyarticular pattern of PsA (p=0.08). Additionally our study revealed that the HLA-C(∗)02 allele was more frequently observed in PsA patients (OR=5.40, p<0.0005) and also that the HLA-E(∗)01:01 allele was significantly over-represented among HLA-C(∗)02-negative patients in comparison to healthy individuals (OR=6.44, p=0.045). Therefore these results suggest that the HLA-E and HLA-C(∗)02 molecules may also play an important role in determination immune response contributing to the PsA development.

Cytokine profiles in axial spondyloarthritis.

Objectives Current studies concentrate on the cytokine network and its role in the pathogenesis of spondyloarthritis (SpA). In this study, we analyzed whether the serum cytokine profile (interleukins: IL-10, IL-11, IL-12, IL-15, IL-17, IL-23 and IL-33) correlates with demographic data, clinical manifestations, disease activity and treatment outcome in a group of patients with axial spondyloarthritis. Material and methods Forty-nine patients with an established diagnosis of axial spondyloarthritis (aSpA) and 19 healthy volunteers as controls were enrolled in the study. Clinical evaluation included patients medical history, 44 joint count, back pain intensity and global disease activity in the preceding week (VAS), the duration of morning stiffness and blood tests. Disease activity was assessed using BASDAI and ASDAS-CRP. Serum concentration of IL-10, IL-11, IL-12, IL-15, IL-17, IL-23 and IL-33 was determined. Results In patients with aSpA, elevated serum concentration of IL-10, IL-15, IL-17 and IL-23 was detected. In the aSpA group we detected higher values of serum concentration of IL-23 and IL-33 in the subgroup with anterior uveitis (83.1 ±184.0 pg/ml vs. 14.0 ±17.1 pg/ml, p < 0.0001 and 45.5 ±71.9 pg/ml vs. 18.4 ±14.3 pg/ml, p < 0.0001, respectively). Additionally, in the subgroup with peripheral arthritis, elevation of serum concentration of IL-12 (249.3 ±246.9 pg/ml vs. 99.9 ±105.9 pg/ml, p = 0.0001) was detected. Patients with preradiological SpA had higher serum concentration of IL-17 than patients with established diagnosis of AS (6.37 ±8.50 pg/ml vs. 2.04 ±2.98 pg/ml, p = 0.0295). No differences in serum concentration of analyzed cytokines were found between the subgroup with low to moderate disease activity and the subgroup with high to very high disease activity. Conclusions We report that in aSpA patients, compared to controls, elevated serum concentrations of IL-10, IL-15, IL-17 and IL-23 were observed. Some cytokines may predispose to a more severe course of aSpA.

Interstitial lung disease in systemic sclerosis: challenges in early diagnosis and management

Interstitial lung disease (ILD) is a group of lung diseases characterized by thickening of the interstitium surrounding pulmonary alveolar walls. It is related to specific radiographic features in lung imaging and/or the presence of restrictive disorders in pulmonary function tests (PFTs). ILD is one of the leading causes of death in systemic sclerosis patients. Major risk factors of ILD associated with SSc (SSc-ILD) include male sex, diffuse type of cutaneous SSc and presence of anti-Scl-70 antibodies. SSc-ILD is challenging to diagnose at an early stage as the symptoms are non-specific. The greatest risk of its development is during the 4–5 years after the initial diagnosis of systemic sclerosis. Clinical vigilance at the time, including regular pulmonary function tests and/or high-resolution com-puted tomography (HRCT), is needed. The aim of this paper is to summarize the current knowledge on early diagnostic methods and progression risk factors for SSc-ILD.

AB0006 Mir-26a polymorphism is associated with susceptibility of rheumatoid and psoriatic arthritis

Background Serum levels of miR-26a has been reported to act as potential biomarker of rheumatic diseases. Objectives The aim of the study was to analyse the genetic variation and expression of miR-26a as potential diagnostic and/or prognostic markers of rheumatoid diseases. Methods The miR-26a polymorphism was examined in 111 patients with rheumatoid arthritis (RA), 86 patients with psoriatic arthritis (PsA) and 162 healthy blood donors that served as a control group. Genotyping for miR-26a rs7372209 was performed using a LightSNiP assay. For analysis of the miR-26a expression, RNA was isolated from sera of 15 RA patients (before and 3 months after anti-TNF treatment) and 10 controls (NucleospinmiRNA Plasma; MACHEREY-NAGEL GmbH and Co. KG) followed by cDNA synthesis (TaqMan MicroRNA Reverse Transcription Kit; Applied BiosystemsTM by Life Technologies) and Real-time PCR amplifications with hsa-miR-26a TaqMan specific and U6 snRNA control primers for each probe. The results were analysed using the (ΔΔCt) calculations. Results It was found that the presence of miR-26a TT genotype (rs7372209) more than 5 times increases the risk of RA (OR=5.28, p=0.003) while the presence of CC homozygotes is associated with the risk of PsA (OR=1.77, p=0.037). There was no significant difference in the miR-26a serum levels between patients and controls. Also miR-26a serum levels did not significantly differed between RA patients before, 3 and 6 months after the implementation of biological therapy with TNF-alpha inhibitors. Conclusions These results imply that miR-26a rs7372209 allelic variants differentially affect the risk of rheumatoid and psoriatic arthritis while anti-TNF biological treatment seems not to affect the miR-26a expression in RA patients. Disclosure of Interest None declared

FRI0474 Serum Level of IL-23 and IL-23R Polymorphisms in Patients with Psoriatic Arthritis

Background Interleukin (IL) - 23 is one of the of cytokines involved in systemic inflammation. Interaction between this cytokine and its receptor (IL-23R) that plays an important role in pathogenesis of psoriatic arthritis (PsA). Objectives The present study aimed to assess the associations between polymorphisms within gene coding IL-23R, IL-23 serum levels and disease activity in patients with PsA. Methods Fifty-two PsA patients (diagnosed by the criteria recommended by CASPAR group) were genotyped for the IL-23R (rs11209026 and rs7530511) polymorphisms. The nuclear factor kappa (NF-kB1 (rs28362491; ins/del)) polymorphism (associated with the promoter activity of this gene and cytokine gene expressions, including IL-23) was also analyzed in PsA patients group. IL-23 serum levels were assessed by ELISA in patients with PsA, and for comparison, 10 healthy individuals. These laboratory data were further related with clinical characteristics of the patients. Disease Activity Score was measured (swollen and tender joints, ESR, CRP) in addition to BASDAI, BASFI, VAS, and PASI scores. Results Significantly (p<0.05) elevated levels of IL-23 cytokine were observed in PsA patients (126.5 pg/ml) when compared to control group (24.9 pg/ml). Moreover, IL-23 serum levels were associated with the IL-23R rs7530511 polymorphism. Patients carrying the IL-23R T allele characterized with higher concentrations of IL-23 in serum (299.1 vs 86.8, p<0.05). Interestingly, patients with the IL-23R T allele were also more frequently carrying the ins/ins homozygous NF-kB1 genotype (associated with a better promoter activity and higher expression of cytokines) (7/17 vs 4/34, distribution of the T allele among ins/ins vs del allele positive patients, p=0.03). No association was found between for IL-23 levels or IL-23R polymorphisms and disease activity. Conclusions Patients with PsA characterize with higher serum levels of IL-23 than healthy individuals. IL-23 concentrations in serum of PsA patients are associated with the polymorphism (rs7530511) of IL-23 receptor encoding gene. Disclosure of Interest None declared

AB0762 Clinical Associations of the IL-17A, IL-17F and Nfkb-1 Gene Polymorphisms in Patients with Rheumatoid and Psoriatic Arthritis

Background The pro-inflammatory cytokines IL-17A and IL-17F, primarily produced by Th17 cells, has been shown to be involved in all stages of the autoimmune and inflammatory diseases [1]. Th17 cells are thought to play an important role in rheumatoid (RA) and psoriatic arthritis (PsA) [2]. Objectives The present study aimed to determine the association between their polymorphic variants with possible involvement in disease susceptibility and effect on disease progression in patients with PsA and RA susceptibility, progression and response to therapy with TNF-alpha inhibitors. Methods Eighty-nine RA and 54 PsA patients and 126 healthy individuals were investigated and genotyped for the IL-17A (rs2275913; G-197A), IL-17F (rs763780; A7488G; His161Arg) and transcription nuclear factor NFkB-1 (rs28362491; -94 ins/del ATTG) alleles. Results Female RA patients carrying the IL-17A wild type genotype more frequently presented with radiographic changes grade 4 (Steinbrocker) (8/24 vs. 6/47, p=0.058) and were characterized by more active disease after 3 months of therapy with the TNF inhibitors (12/23 vs. 15/45, p=0.040). The presence of the IL-17F minor (G) variant (OR=3.97, p<0.001) and its homozygosity (OR=29.62, p<0.001) was more frequent among RA patients than healthy individuals. The NFkB1 del genotype was more frequently observed in patients with PsA onset over 40 year of age (16/20 vs. 8/17, p=0.047). PsA patients lacking this NFkB1 del/del genotype (ins positive cases) more frequently presented with severe disease (PASI >10, BSA >10, more than five swollen and tender joints) and subjected to anti-TNF-alpha therapy (20/21 vs. 12/18, p=0.035). Conclusions These results suggest that the IL-17A and IL-17F polymoprhisms play a significant role in RA while NFkB-1 variants seem to affect PsA. References Miossec P, Kolls JK. Targeting IL-17 and TH 17 cells in chronic inflammation. Nat Rev Drug Discov 2012; 11:763:776 Shen H., Goodal JC, Hill Ga ston JS. Frequency and phenotype of peripheral blood Th17 cells in ankylosing spodylitis and rheumatoid arthritis. Arthritis Rheum 2009; 60:1647-1656. Disclosure of Interest None declared DOI 10.1136/annrheumdis-2014-eular.5856

AB0763 Serum Level IL-23, IL-18, IL-17A, IL-17F, IL-12, IL-6 in Psoriatic Arthritis PSA Patients Hospitalized at the Rheumatology Clinic of Medical University in Wroclaw, Poland

Background IL-23, IL-12, IL-18, IL-17, IL-6 belong to the family of cytokines involved in systemic inflammation which plays a key role in the pathogenesis of psoriasis (PSO) and psoriatic arthritis (PsA).[1,2] Objectives The aim of this study was to investigate the association between serum levels of IL-23, IL-12, IL-18, IL-17, IL-6, and disease activity in PsA patients and control group. Methods The study group consisted of 55 patients with PsA (32 men and 23 women) hospitalized at the Rheumatology Clinic of Medical University in Wroclaw, Poland, between 2012 and 2013. Ten (n=10) healthy persons served as a control group. Study was performed with the permission of the Commission of Bioethics (Medical University, Wroclaw, Poland). The median age of patients was 48 years. PsA was diagnosed by the criteria recommended by CASPAR group. The average duration of PsA and PSO was 7 and 15 years, respectively. Serum levels of cytokines were assessed with ELISA test. Disease Activity Score was measured (swollen and tender joints, ESR, CRP) in addition to BASDAI, BASFI, VAS, and PASI scores. Results Mean serum levels of cytokines are depicted in the table below: Mean serum levels of IL in PsA Mean serum levels of IL in control group Threshold [pg/ml] [pg/ml] [pg/ml] Il-23 126,5 24,9 <63,57 Il-12 123,7 1,8 65–483 Il-6 7,7 0,24 <12,5 Il-17A 3,35 0,36 <3 Il-17F 4,8 0 <6 Il-18 164,87 59 <393 We observed elevated levels of IL-23 cytokine (126,5 pg/ml) in PsA patients, which reached statistical significance (P<0.05) if compared to control group (24,9 pg/ml). There were no significant differences between groups in other cytokines. We found a positive correlation between IL-6 serum levels and both OB and CRP. Conclusions In our study we have successfully demonstrated a significant increase in IL-23 serum levels in PsA patients. In addition we identified a possible link between serum levels of IL-6 and parameters of inflammatory state. These data suggest a potential role of IL-23 and IL-6 in pathogenesis of PsA. References Gottlieb A., Narang K. Ustekinumab in the treatment of psoriatic arthritis: latest findings and clinical potential. Therapeutic Advances in Musculoskeletal Disease 2013; 5(5) 277-285 Her M., Kavanaugh A. Treatment of spondyloarthropathy: the potential for agents other than TNF inhibitors. Curr Opin Rheumatol 2013, 25:455-459 Disclosure of Interest None declared DOI 10.1136/annrheumdis-2014-eular.5815

THU0404 Vitamin D level in patients with early arthritis. A preliminary report

Background Vitamin D, because of its pleiotropic effect, is involved in many aspects of pathophysiology including calcium-phosphorus metabolism, cellular growth and differentiation and immune functions. Epidemiological studies suggest that deficiency of vitamin D may be a risk factor of many conditions including autoimmune diseases such as rheumatoid arthritis or systemic lupus erythematosus [1,2]. There are reports showing an inverse relationship between vitamin D concentrations and measures of disease activity in early inflammatory arthritis [3]. But some other data are conflicting [4]. Objectives To estimate the vitamin D status in patients with early arthritis and to find relationship with some relevant features of disease activity. To examine associations between 25(OH)D level and current treatment. Methods The study participants were patients with the diagnosis of early arthritis (lasting <2years) hospitalized in the Dept. of Rheumatology University Hospital, Wroclaw, Poland. 25(OH)D was measured in serum samples using commercial ELISA kit, all the necessary data concerning disease activity were also collected. Functional status was derived from HAQ questionnaire. Patients were diagnosed as having vitamin D deficiency when serum level of 25(OH)D were under 30 ng/ml. Results A total of 39 patients (31 women, 8 men,), mean age 45.2±16.8 years were included in the study. In analyzed group mean disease duration was 6.3±7.3 months. Estimated disease activity according to DAS28 had a mean value 4.8±1.4. Mean serum level of 25(OH)D was 18.1±9.4 ng/ml. Only 2 out of 39 patients had 25(OH)D level over 30 ng/ml, the rest had under 12 ng/ml (severe deficiency). There was no correlation between vitamin D level and parameters of disease activity (ESR, C reactive protein, DAS28, morphologic parameters of the peripheral blood) or functional status. There was also no differences in 25(OH)D serum concentrations among women and men. Analyzing the influence of the treatment on 25(OH)D levels, the levels was significantly higher in patients treated with sulphasalazin, compared to those not treated with any of the disease modifying antirheumatic drugs. The use of corticosteroids (mean dose of prednisone in the study group was 10.4±8.6 mg per 24h) did not influence the 25(OH)D serum level significantly. Conclusions The prevalence of vitamin D deficiency among patients with early arthritis is very high. The appropriate treatment regiments seem to influence the serum 25(OH)D levels. In contrast to earlier reports with recent-onset inflammatory arthritis, in our study, there were no associations between 25(OH)D levels and disease activity. References Fletcher JM, Basdeo SAet al. Therapeutic use of vitamin d and its analogues in autoimmunity.Recent Pat Inflamm Allergy Drug Discov.2012 Jan 1;6(1):22-34. Cutolo M, Pizzorni C, Sulli A. Vitamin D endocrine system involvement in autoimmune rheumatic diseases.Autoimmun Rev.2011 Dec;11(2):84-7. Patel S, et al. Association between serum vitamin D metabolite levels and disease activity in patients with early inflammatory polyarthritis. Arthritis Rheum. 2007; 56(7):2143–9. Craig SM, Yu F, Curtis JR, Alarcόn GS et al. Vitamin D status and its associations with disease activity and severity in African Americans with recent-onset rheumatoid arthritis.J Rheumatol.2010 Feb;37(2):275-81. Disclosure of Interest None Declared

Early diagnostic Achilles tendinopathy in patients with psoriasis and arthralgia

Psoriatic arthritis belongs to a group of spondyloarthropaties that are characterised by, among other things, enthesopathy. The most typical enthesitis is heel pain related to inflammation of the Achilles tendon or the plantar fascia insertion. Musculoskeletal ultrasonography is a useful tool for evaluating joint and soft tissue pathology and it can be used to visualise enthesopathy.1,2,3 …