872 Scientific Abstracts inflammation, and fibrosis. Microvascular injury and insufficient angiogenesis lead to tissue ischemia and clinical manifestations such as fingertip ulcers. UCD-206 chickens are the only animal model showing all hallmarks of the human disease, including microvascular damage and insufficient angiogenesis. The major inducer of angiogenesis is vascular endothelial growth factor (VEGF). Uncontrolled expression of VEGF and its receptors VEGFR-1 and VEGFR-2 has been shown in both, human and avian SSc. Another set of molecules that critically regulate angiogenesis are the endothelial cell-specific receptor tyrosine kinase 2 (Tie-2) and its ligands angiopoietin-1 (Ang-1) and angiopoietin-2 (Ang-2). Although abnormal plasma levels of soluble sTie-2, sAng-1, and sAng-2 have been reported in SSc patients, expression in lesional tissues has not been examined. Objectives: To study the expression of Tie-2, Ang-1 and Ang-2 in lesional and non-lesional skin of UCD-206 chickens. Methods: The expression of Tie-2, Ang-1 and Ang-2 was studied by indirect immunofluorescence tests on frozen tissue sections from lesional and non-lesional neck skins from 5 weeks old UCD-206 chickens. Age matched H.B15 chickens were used as healthy controls. Samples were double stained with monoclonal anti-Tie-2 antibody and the endothelial cell marker anti-von Willebrand factor (vWF), monoclonal anti-Ang-1 antibody and anti-alpha smooth muscle actin antibody, as a marker for mural cells, or with monoclonal anti-Ang-2 antibody and anti-vWF. The tissue cytometry software TissueQuest ® was used for quantitative analyses. Statistical significance was calculated using the Mann-Whitney-U test. Results: The number of Tie-2 expressing endothelial cells was significantly increased in lesional skin compared to non-lesional UCD-206 skin and healthy controls. Expression of Ang-2 was significantly decreased in lesional compared to non-lesional UCD-206 skin and to healthy H.B15 skin. Whereas no significant difference was seen in the numbers of Ang-1 expressing mural cells, other Ang-1 expressing cells were reduced in lesional skin compared to non-lesional and healthy controls. The altered endothelial expression of Ang-2 and Tie-2 resulted in significantly decreased Ang-2:Tie-2 ratios in lesional (median 0.9, IQR 0.3 – 1.8) compared to non-lesional UCD-206 skin (median 6.9, IQR 3.4 – 14.2, p≤0.0005) and healthy controls. (median 11.2, IQR 2.4 – 25.2, p≤0.0005). The Ang-2:Ang-1 ratio was also significantly diminished in lesional (median 0.4, IQR 0.1 – 0.5) compared to non-lesional UCD-206 skin (median 0.7, IQR 0.4 – 1.3, p≤0.01) and healthy controls. (median 1.0, IQR 0.6 – 1.6, p≤0.002). Conclusions: Altered expression of Ang-1, Ang-2 and Tie-2 might contribute to the vasculopathy in SSc. Acknowledgements: This work was funded by the Austrian Science Fund (FWF): P23230-B13. Disclosure of Interest: None declared DOI: 10.1136/annrheumdis-2014-eular.2338 AB0207 ENHANCED FORMATION AND IMPAIRED DEGRADATION OF NEUTROPHIL EXTRACELLULAR TRAPS IN DERMATOMYOSITIS AND POLYMYOSITIS: A POTENTIAL CONTRIBUTOR TO DM/PM- COMPLICATED ILD S. Zhang , G. Wang, X. Lu. Rheumatology, China-Japan Friendship Hospital, Beijing, China Background: Dermatomyositis (DM) and polymyosits (PM) are systemic au- toimmune diseases with ambiguous pathogenesis. Neutrophil extracellular traps (NETs) is considered to play an important role in pathogenesis of autoimmune disease. Objectives: The present study tested the hypothesis that NETs may be pathogenic in DM/PM. Methods: Plasma samples from 97 DM/PM patients (72DM, 25PM) and 54 healthy controls were tested for formation and degradation of NETs. Plasma DNase I activity was tested to further analyze the reason for abnormal degradation of NETs. Results from thirty five DM patients and seven PM patients with ILD were compared with DM/PM patients without ILD. Results: Compared with Control, DM/PM exhibited a significantly enhanced NETs formation (191.6±52.88 RFUs vs. 246±93.48 RFUs, P=0.002), which was supported by elevated level of plasma LL-37 and cfDNA in DM/PM. DM/PM also exhibited a significantly decreased ability to degrade NETs and DNase I activity. The percentage of NETs degraded by plasma significantly correlated with plasma DNase I activity in DM/PM. Moreover, DM/PM patients with ILD exhibited a significantly lower ability to degrade NETs and DNase I activity than patients without. DNase I activity in patients with anti-Jo-1 antibodies was significantly lower than that in patients with negative anti-Jo-1 antibodies. Glucocorticoids therapy seems to improve the activity of DNase I. Conclusions: The collective evidence demonstrated that the excessively formed NETs cannot be completely degraded owing to impairment of DNase I activity in DM/PM, especially in DML/PML, suggesting that residual NETs may participate in pathogenesis of ILD and DNase I may be a potential target. References: [1] Lundberg IE. Pathogenesis, classification and treatment of inflammatory myopathies. Nat Rev Rheumatol 2011;7:297–306 [2] Danoff SK, Casciola-Rosen L. The lung as a possible target for the immune reaction in myositis. Arthritis Research & Therapy 2011;13:230. [3] Cheng OZ, Palaniyar N. NET balancing: a problem in inflammatory lung diseases. Front Immunol 2013;4:1. [4] Khandpur R, Carmona-Rivera C, Vivekanandan-Giri A, et al. NETs Are a Source of Citrullinated Autoantigens and Stimulate Inflammatory Responses in Rheumatoid Arthritis. Sci Transl Med 2013 Mar 27;5(178):178ra40. [5] Keshari RS, Jyoti A, Dubey M, Kothari N, Kohli M, Bogra J, Barthwal MK, Dikshit M. Cytokines Induced Neutrophil Extracellular Traps Formation: Implication for the Inflammatory Disease Condition. PLoS ONE 2012;7(10): e48111. Acknowledgements: The authors thank all patients and volunteers who partici- pated in this study. Disclosure of Interest: None declared DOI: 10.1136/annrheumdis-2014-eular.2472 AB0208 FMD, VEGF, AND IL-6 IN VEDOSS PATIENTS: THE PRECOCITY OF ENDOTHELIAL DYSFUNCTION S.L. Bosello 1 , G. Canestrari 1 , A. Capacci 1 , A. Di Giorgio 2 , G. De Luca 1 , M. Bocci 1 , F. Parisi 1 , G. Berardi 1 , M. Rucco 1 , S. Alivernini 1 , A. Santoliquido 2 , G. Ferraccioli 1 . 1 Division of Rheumatology, Institute of Rheumatology; 2 Division of Angiology, Institute of Internal Medicine, Rome, Italy Background: Endothelial dysfunction is a key feature of systemic sclerosis (SSc) and the involvement of the microvasculature is one of the earliest features of the disease. Recent new criteria for very early diagnosis of systemic sclerosis (VEDOSS) have been proposed. Objectives: The aim of this study was to investigate brachial artery endothelial- dependent flow-mediated dilation (FMD), IL-6 and VEGF levels in patients with primary Raynaud’s phenomenon (RP) and SSc. Methods: In this study we enrolled 59 patients: 10 VEDOSS patients fulfilling the proposed VEDOSS criteria, 28 SSc patients fulfilling the 1987 ACR criteria, 11 gender and age matched healthy individuals as first control group and 10 gender and age matched primary RP patients with normal capillaroscopic findings as the second control group. Demographic, clinical and immunological parameters have been collected at the beginning of the study. Ultrasound assessment of FMD was performed in all RP subjects and in healthy subjects to evaluate endothelial dysfunction. VEGF, VEGF-RII, IL-6 and IL-6R plasma were determined by ELISA. Results: Scleroderma patients showed a reduced FMD (5.8±4.7%) com- pared to healthy controls (18.9±6.7%) (p<0.0001), and to subjects with RP (1.6±5.6%),(p=0.001). FMD of scleroderma patients and VEDOSS patients was comparable (6.6±3.8%). VEDOSS patients showed values of FMD significantly compromised compared to those of RP (p=0.05) and healthy controls (p<0.001). Finally, patients with primitive RP showed reduced FMD values compared to healthy controls (p=0.008). The values of FMD correlated inversely with disease duration (r=-0.46, p=0.004) and directly with the levels of KCO (r=0.47, p=0.007). The plasma levels of VEGF in patients with SSc (29.8±40.7 pg/ml) were signifi- cantly higher than in healthy controls (13.0±24.8 pg/ml,(p=0.03) while they were similar to those of patients with VEDOSS (22.6±21.7 pg/ml). Plasma levels of IL-6 were higher in SSc patients (5.7±11.1 pg/ml) compared to VEDOSS (1.5±1.2 pg/ml) (p=0.04),RP (1.1±0.5 pg/ml), (p<0.0001) and healthy controls (1.9±3.1 pg/ml) (p<0.001). Levels of VEGF-R2 and sIL-6R were comparable in all groups. IL-6 levels were higher in dcSSc patients than in lcSSc patients (7.9±13.0 pg/ml vs 1,9±0,6 pg/ml) (p=0,0027). Finally in patients with SSc the capillaroscopic density inversely correlated with the levels of IL-6, the Activity and Severity Indices, the Skin Score and the duration of the disease. Conclusions: An impairment of FMD was present in patients with RP, in particular in SSc and VEDOSS patients, suggesting a contemporary impairment of microvascular and macrovascular compartments. The deeper FMD impairment that characterize either SSc and VEDOSS patients, suggests that the endothelial dysfunction is already established since the early phases of the disease and it worses during the disease. Disclosure of Interest: None declared DOI: 10.1136/annrheumdis-2014-eular.4446 AB0209 ENDOTHELIN-1 INDUCES DIRECT ACTIVATION OF JNK AND C-JUN SIGNALLING PATHWAYS IN CULTURED HUMAN DERMAL FIBROBLASTS S. Soldano 1 , R. Brizzolara 1 , P. Montagna 1 , A. Sulli 1 , E. Gianetta 2 , M. Cutolo 1 . 1 Research Laboratory and Academic Division of Clinical Rheumatology, Department of Internal Medicine; 2 Department of Surgical Sciences and Integrated Diagnostics, University of Genova, Genova, Italy Background: Myofibroblasts are final key mediators of the fibrotic process in several autoimmune connective tissue diseases, including systemic sclerosis (SSc) and contribute to the excessive synthesis and deposition of extracellular matrix (ECM) molecules (i.e. fibrillar collagens, fibronectin) in skin and internal organs (1-3). Several growth factors, such as transforming growth factorβ (TGFβ), induce myofibroblast differentiation through the activation of intracellular signalling pathways, as Jun-N-terminal kinase (JNK) or c-Jun (4). Endothelin-1 (ET-1) plays an important role in SSc, inducing the activation of myofibroblasts to enhance the synthesis of ECM proteins (5,6). Objectives: To investigate the ability of ET-1 to directly induce the activation of the intracellular signalling pathways involved in the profibrotic myofibroblast activation in cultured human normal dermal fibroblasts.