E.Y. Lee

Renal expression of galectin-3 in systemic lupus erythematosus patients with nephritis

The aim of the study is to characterize the expression pattern of galectin-3 (Gal-3) in renal tissues of patients with systemic lupus erythematosus (SLE) nephritis and to determine whether tissue and serum Gal-3 are associated with SLE nephritis. Gal-3 expressions were examined with immunohistochemistry in renal biopsy specimens of 88 patients with SLE nephritis and in five normal specimens. Activity and chronicity indexes and glomerular Gal-3 expressions were analysed in each specimen. Serum Gal-3 levels were measured using enzyme-linked immunosorbent assays in 20 patients with SLE, including 11 with nephritis, and in 50 healthy controls. Glomerular Gal-3 expression was observed in 81.8% (72/88) of patients with SLE nephritis but not in 5 controls. Gal-3 staining was attributed mainly to its cellular expression rather than its deposition, and Gal-3 expression levels were correlated with histologic activity indexes, anti-dsDNA titers, and complement 3 and 4 levels. Serum Gal-3 levels were higher in patients with SLE, particularly in those with nephritis, than in healthy controls, and correlated with anti-dsDNA titers. In conclusion, glomerular Gal-3 expression in renal tissue and serum Gal-3 levels were elevated in patients with SLE nephritis versus healthy controls; moreover, they reflected disease activity. These findings suggest that Gal-3 might contribute to the inflammatory process in SLE.

Flow cytometric assessment of anti-neuronal antibodies in central nervous system involvement of systemic lupus erythematosus and other autoimmune diseases

The objective of this study is to evaluate the association between anti-neuronal antibody (anti-NA) and central nervous system (CNS) manifestations of systemic lupus erythematosus (SLE) and other rheumatic diseases using a flow cytometric method. Anti-NA was measured by flow cytometry in serum and cerebrospinal fluid (CSF) samples from patients with SLE (n = 44 for serum, n = 17 for CSF), other rheumatic diseases (n = 64 for serum, n = 21 for CSF) and from healthy controls (n = 65 for serum, n = 18 for CSF). Serum anti-NA was more frequently observed in SLE (31.8%, 14/44) than in other rheumatic diseases (4.7%, 3/64, P < 0.001) or in healthy controls (0%, 0/65, P < 0.00001). In SLE patients, the frequency of serum anti-NA was significantly higher in CNS-SLE (76.5%, 13/17) than in non CNS-SLE (3.7%, 1/27, P < 0.000001). CSF anti-NA was detected in 88.2% (15/17) of CNS-SLE and was more frequently detected in CNS-SLE (15/17, 88.2%) than in other rheumatic diseases with CNS involvement (1/21, 4.8%, P < 0.000001) or in healthy controls (0/18, P < 0.000001). In conclusion, serum anti-NA was more frequently found in CNS-SLE than in non CNS-SLE, other rheumatic diseases or in healthy controls. The frequency of CSF anti-NA in CNS-SLE was significantly higher than in other rheumatic diseases with CNS involvement or in healthy controls. Lupus (2008) 17, 21—25.

FRI0487 Association between Fever Pattern and Clinical Manifestations in Adult Onset of Still's Disease: Unbiased Analysis of Fever Pattern Using Hierarchical Clustering

Background Adult onset of Stills disease (AOSD) is a systemic inflammatory disease characterized by fever, arthritis, salmon pink rash and elevated serum inflammatory markers. The classical fever of AOSD is intermittent with a quotidian or double-quotidian pattern. However, a significant subset of AOSD exhibits persistently high fever with little temperature fluctuation. It remains unclear as to whether the certain fever pattern is associated with specific clinical manifestations. Objectives To investigate the association between fever pattern and clinical characteristics in patients with AOSD. Methods A total of 70 patients with AOSD who were treated as inpatient at Seoul National University Hospital from 2004 through 2015 were enrolled. Patients were grouped based on the fever curves on days 2, 3 and 4 using hierarchical clustering. The clinical and laboratory characteristics of the groups were compared. Results 70 patients were divided into 2 groups based on the fever curves. The group 1 with 14 patients had a higher mean temperature (38.1±0.4°C vs. 37.2±0.5°C, p<0.001) with wider daily variation (2.7±0.9°C vs. 1.9±0.7°C, p<0.001) as compared to 56 patients in the group 2. Group 1 tended to be younger (38.4 ± 14.7 years vs. 46.1 ± 17.6 years, p=0.141) (Figure). Fever, arthritis, arthralgia and rash did not differ between them. However, group 1 tended to have more pericardial and lung involvement (42.9% v.s 23.2%, p=0.181). Group 1 had significantly lower platelet count (198,900 ± 68,000/μl vs. 312,900 ± 156,900/μl, p=0.0001), higher LDH (816.6 ± 376.4 IU/L vs. 477.5 ± 327.1 IU/L, p=0.002), higher mean ferritin level (27,004.1 ± 48,891.5 ug/mL vs. 6,852 ± 10,628.2 ug/mL, p=0.072) and d-dimer (9.5 ±7.9 ug/mL vs 3.3 ± 3.4 ug/mL) as compared to group 2, whereas white blood cell count, hemoglobin levels, CRP levels did note differ between the both groups. Group 1 tended to require longer time to a clinical remission (455.5 ± 850 days vs. 9.4 ± 115.7 days, p=0.137). Conclusions The unbiased analysis of fever reveals the presence of at least 2 distinctive fever patterns in AOSD. Spiking fever with higher daily variation was more often associated with higher inflammatory markers and coagulopathy and required longer treatment. Thus, fever pattern might be a prognostic factor in AOSD and AOSD patients with spiking fever might need more intensified treatment. Disclosure of Interest None declared

OP0094 Survival Benefit of Early Use of Cyclosporine in Dermatomyositis-Associated Interstitial Lung Disease

Background Interstitial lung disease (ILD) is the most common cause of mortality in patients with dermatomyositis (DM). Cyclosporine has been reported to improve clinical outcome in some patients with DM-associated ILD. Objectives To investigate the benefit of early introduction of cyclosporine on the survival of patients with DM-associated ILD. Methods All the patients with DM-associated ILD, who were treated with cyclosporine at Seoul National University Hospital (SNUH) and Seoul National University Bundang Hospital (SNUBH) between 1990 and 2013, were enrolled. Enrolled patients were diagnosed with DM according to Bohan-Peters classification criteria (n=28) or with clinically amyopathic dermatomyositis (CADM) according to Sontheimers classification criteria (n=19). ILD was diagnosed based on the typical findings on computed tomography (CT) and relevant respiratory symptoms or signs. Kaplan-Meier survival analysis was applied to compare overall mortality rates between patients who took cyclosporine as the initial treatment and those as the delayed treatment. Cox regression analysis was used to estimate the benefit of early treatment with cyclosporine after adjusting confounding characteristics. Results Among the 47 patients who were diagnosed with DM-associated ILD, 16 patients (34.04%) received cyclosprorine initially after diagnosis and 31 patients (65.96%) received cyclosporine after the trial of other immunosuppressants such as corticosteroids, cyclophosphamide or azathioprine. The mean time of follow-up was 43.57 person-years in initial treatment group and 76.23 person-years in delayed treatment group. The mortality rate was significantly lower in the initial treatment group than the delayed treatment group (0.02 person-years vs 0.18 person-years, p=0.0092 by log-rank test). The two groups did not differ in regard to age, sex, duration of disease, presence of autoantibodies, degree of dyspnea at initial visit, initial requirement of oxygen supplement, functional vital capacity (FVC) and diffusion capacity (DLCO) on pulmonary function test (PFT) and the presence of malignancy. Only the proportion of patients with CADM was higher in the initial treatment group than in the delayed treatment group (62.53% vs 29.03%, p=0.027). Survival benefit in the initial cyclosporine treatment remained significant even after adjusting for the CADM status (HR 0.075, 95% CI 0.009 - 0.603, p=0.015 by Cox-regression analysis). Figure 1. Survival curve in initial and delayed use of cyclosporin in patients with dermatomyositis-associated interstitial lung disease. Conclusions This study showed significant survival benefit of initial cyclosporine treatment in patients with DM-associated ILD. A randomized controlled study is warranted to clearly demonstrate the therapeutic benefit of early use of cyclosporine in this potentially fatal disease. Disclosure of Interest None declared DOI 10.1136/annrheumdis-2014-eular.3718

AB0122 The Effect of GV1001, a Peptide Vaccine, in Animal Model of Rheumatoid Arthritis

Background GV1001 is a 16-mer peptide developed as a cancer vaccine to prime the immune system to recognize telomerase. Besides its role as a cancer vaccine, GV1001 has anti-inflammatory effects in in-vitro experiments using macrophage or monocyte. Objectives We evaluated the effect of GV1001 to inhibit joint inflammation in animal model of RA. Methods 7-9 week old BDA/1 mice (Harlan, Netherlands) were used to induce collagen induced arthritis (CIA). Mice were immunized with bovine type II collagen on day 1 and boosted on day 21. Treatments with GV1001 (0.2 nM, 1 nM, 2 nM, 5 nM, and 10 nM) or placebo were started on day 21, three times in alternative week (on day 21, 23, 25 and day 35, 37, 39). Weight and arthritis index of the mice using a standardized grading scale (0–3) were monitored from day 21. Observation finished on day 42, harvesting mouse serum for further analysis. The second experiment was proceeded with 0.2 nM GV1001 group, 0.2 nM control peptide and arthritis control without treatment in the same schedule to harvest splenocytes. Control peptide was one of the scrambled peptides of GV1001, consist of the same amino acids with a different sequence (PEP164; PKRPSFIEALTRPLR). Results Mean arthritis index lower tendencies in 0.2 nM GV1001 group compared to placebo group without statistical significance (p=0.1). Serum IL-6 level was significantly lower in 1 nM and 0.2 nM GV1001 group (p<0.01, both) compared to placebo group. In-vitro T cell re-stimulation with collagen, the level of interferon-gamma in 0.2 nM GV1001 treated group was significantly lower than placebo group (p<0.001) and control peptide group (p<0.01). Conclusions Protective effect of GV1001 was not dose dependent and suspected to be related with T cell immune reaction in animal models of RA. These results suggest that GV1001 can modulate immune response in RA patients and has a potential to be used as immunotherapy, such as an RA vaccine. Acknowledgements KAEL-GemVax Co., Ltd provided GV1001 peptide and educational grant KAEL-GemVax had no role in the study design and conduct of the study; the collection, management, analysis, and interpretation of the data. Disclosure of Interest None declared

AB0356 Comparison of the Clinical Characteristics of Rheumatoid Arthritis Patients with and Without Interstitial Lung Disease

Background The clinical importance of rheumatoid arthritis-associated interstitial lung disease has been increased because the usual interstitial pneumonia (UIP) is the most common subtype of interstitial lung disease (ILD) on high-resolution computed tomography (HRCT) in rheumatoid arthritis (RA)and has poor survival rate compared to other forms of ILD related to connective tissue disease. Objectives The clinical characteristics and laboratory data of RA patients with and without ILD were compared in order to clarify the risk factors for ILD in RA patients. Furthermore, risk factors of mortality in the RA patients with ILD group were analyzed. Methods A total of 77 RA patients with ILD and 231 age/sex-, and disease duration-matched RA patients without ILD who were followed from 1991 to 2011 at Seoul National University Hospital were enrolled in this study. Epidemiologic, clinical, and laboratory information including rheumatoid factor (RF) and anti-cyclic citrullinated peptide (anti-CCP) were obtained through medical chart review. The erythrocyte sedimentation rates (ESR), C-reactive protein (CRP) levels, presence of erosion on joint X-rays, and mean corticosteroid doses were examined in order to compare RA outcomes between the RA patients with and without ILD. Cox proportional hazard models were used to estimate the risk of mortality in the RA patients with ILD. Results In the RA with ILD group, the titers of the RF and the anti-CCP were significantly higher compared with those in the RA without ILD group (p=0.001 for both). The RA patients with ILD had higher frequency of a history of tuberculosis or nontuberculous mycobacteria (NTM) (p=0.022). In addition, the RA with ILD group exhibited higher levels of CRP at the time of RA diagnosis (p=0.014) and higher ESR (p=0.022) and CRP levels (p<0.001) throughout the 10-year follow-up period. These patients received higher mean daily dose of corticosteroid (p<0.001). In the subgroup analysis of RA patients with ILD, 28 (36.4%) patients died with the mean follow-up duration of 8.7 years. Male patients, high RF titers and UIP subtype on HRCT had significantly worse survival, and those with nonspecific interstitial pneumonia (NSIP) on HRCT had better survival. In the multivariate analysis, a UIP subtype on HRCT and older age at the time of ILD diagnosis were significantly associated with mortality of RA with ILD.Table 1. Clinical characteristics of RA patients with and without ILD. Conclusions The RA patients with ILD had higher RF and anti-CCP titers and baseline CRP levels compared with those without ILD. A UIP subtype on HRCT and older age at the time of the diagnosis of ILD were significantly associated with mortality of RA with ILD. Disclosure of Interest None declared

Urinary vitamin D-binding protein, a novel biomarker for lupus nephritis, predicts the development of proteinuric flare:

Lupus nephritis (LN) is a major complication of systemic lupus erythematosus (SLE). Conventional biomarkers for assessing renal disease activity are imperfect in predicting clinical outcomes associated with LN. The aim of this study is to identify urinary protein biomarkers that reliably reflect the disease activity or predict clinical outcomes. A quantitative proteomic analysis was performed to identify protein biomarker candidates that can differentiate between SLE patients with and without LN. Selected biomarker candidates were further verified by enzyme-linked immunosorbent assay using urine samples from a larger cohort of SLE patients (nu2009=u2009121) to investigate their predictive values for LN activity measure. Furthermore, the association between urinary levels of a selected panel of potential biomarkers and prognosis of LN was assessed with a four-year follow-up study of renal outcomes. Urinary vitamin D-binding protein (VDBP), transthyretin (TTR), retinol binding protein 4 (RBP4), and prostaglandin D synthase (PTGDS) were significantly elevated in SLE patients with LN, especially in patients with active LN (nu2009=u200921). Among them, VDBP well correlated with severity of proteinuria (rhou2009=u20090.661, pu2009<u20090.001) and renal SLE Disease Activity Index (renal SLEDAI) (rhou2009=u20090.520, pu2009<u20090.001). In the four-year follow-up, VDBP was a significant risk factor (hazard ratio 9.627, 95% confidence interval 1.698 to 54.571, pu2009=u20090.011) for the development of proteinuric flare in SLE patients without proteinuria (nu2009=u2009100) after adjustments for multiple confounders. Urinary VDBP correlated with proteinuria and renal SLEDAI, and predicted the development of proteinuria.

AB0071 Alpha-Enolase Expression on The Cell Surface of Osteoclast Precursors Plays A Positive Role in Osteoclastogenesis of Monocyte/macrophage in Rheumatoid Arthritis

Background The local bone loss around the inflamed joints is found from the early stage of rheumatoid arthritis. We previously reported that cell-surface expression of alpha-enolase (ENO1), multifunctional glycolytic protein, is increased in monocytes and macrophages isolated from RA patients and related to production of proinflammatory cytokines (1). Objectives This study was performed to examine the role of surface ENO1 on osteoclastogenesis in rheumatoid arthritis. Methods Monocytes isolated from RA patients and healthy control were used to examine the surface level of ENO1 by flow cytometry and confocal microscopy. U937, human monocytic cell line, was treated with ENO1-targeting siRNA (siENO1) to down-regulate ENO1 expression, then osteoclast differentiation were induced by RANKL and the level of ENO1 and NFATc1 was measured. Also, monocytes from RA patients were differentiated into macrophages and transfected with siENO1 to assess the effect on osteoclast differentiation. Results We found the surface expression level of ENO1 was higher in monocytes isolated from RA patients than healthy control and gradually decreased during osteoclastogenesis in both cells, while the total level of ENO1 remained unchanged. ENO1-specific siRNA significantly decreased the surface ENO1 level in U937, but did not affect the viability and proliferation of the cells. When osteoclast formation was induced by RANKL, osteoclast-specific transcription factor, nuclear factor of activated T-cells, cytoplasmic 1 (NFATc1) expression was markedly inhibited in ENO1 down-regulated cells compared to control cells. We also examined the effect of down regulation of ENO1 in osteoclast precursor cells isolated from RA patients (N=10). The surface ENO1 level was significantly decreased and RANKL-induced osteoclast differentiation was markedly inhibited in ENO1 down-regulated cells compared to control cells. Conclusions These results suggest that increased expression level of ENO1 on the surface of monocytes/macrophages have positive effect on osteoclast formation in RA. References S. Bae, H. Kim, N. Lee, C. Won, H-R. Kim, Y. Hwang, Y W. Song, J S. Kang, and W J. Lee. A-Enolase expressed on the surfaces of monocytes and macrophages induces robust synovial inflammation in rheumatoid arthritis. The Journal of Immunology. (2012) 189: 365–372. Disclosure of Interest None declared

AB0317 Drug Survival of Biologic Agents in Rheumatoid Arthritis Using 10 Years of Data from The National Health Insurance in Korea: A Population-Based Study and Comparison with A Single-Institution Cohort

Background Since 2004, Korean national health insurance began to cover the cost of biologic agents in rheumatoid arthritis (RA) patients experiencing treatment failure by conventional disease modifying antirheumatic agents. However, real-world issues such as selecting or switching among biologic agents in clinical practice became more complicated as types of the agents increased. Objectives To evaluate 1) first choice among biologic agents, 2) drug retention rate and duration and 3) switching rate and duration before switching of each biologic agent in RA. Methods We used prospective cohort data by the National Health Insurance Service (NHIS) in Korea, which consisted of more than one million subjects representing age, sex and income status of whole Korean population followed from 2004 to 2013. All RA patients with any experience of biologic agents were checked by the code of each drug and KCD9 code of diagnosis including all types of RA. To supplement the results of the population based study, we also compared and validated with the independent cohort of biologic agent users in Seoul national university hospital (SNUH). Results One hundred and seventy three patients were found to experience any of TNFa inhibitors from Jan. 2004 to Dec. 2013. Etanercept was the most frequent choice as first TNFa inhibitor (43.4%), followed by adalimumab (35.3%), infliximab (20.2%), golimumab (0.6%) and abatacept (0.6%). Among TNFa inhibitors, retention rates of etanercept (73.1%, 62.7%) at month 12 and 24 were significantly higher (p=0.039, both) than adalimumab (57.7%, 44.2%) and infliximab (45.5%, 36.4%). In SNUH cohort, etanercept (79.8%, 67.0%) also had better retention rate than adalimumab (51.3%, 48.7%) and infliximab (58.6%, 34.5%) at month 12 and 24 respectively. Mean duration of retention was almost twice longer in etanercept (3.42 years) than adalimumab (1.83 years, p=0.004) and infliximab (1.71 years, p=0.011). Similarly, etanercept (3.24 years) prescribed longer than adalimumab (1.98 years) and infliximab (1.63 years) with statistical significance in SNUH cohort. Etanercept also had lower switching rate (20.9%) than adalimumab (32.1%) and infliximab (38.5%, p=0.040) and the tendency were similar in SNUH cohort. Preferred biologic agents after switching were adalimumab (27.4%) and rituximab (24.2%). Etanercept users (1.35 years) had longer duration of use before switching than infliximab (0.60 years, p=0.050) but no significant difference was shown comparing with adalimumab (0.73years, p=0.483). Conclusions As golimumab, abatacept, tocilizumab were available relatively later than existing TNFa inhibitors and rituximab, more follow up period would be required to evaluate their value in treating RA. This research can be a pilot study before analyzing nationwide population cohort encompassing whole population of Korea. Conclusively, etanercept has been prescribed longer in general and the rate of retention till 2 years was also better then adalimumab and infliximab for 10 years after national insurance coverage. Disclosure of Interest None declared

FRI0080 Effects of IL-6 Receptor Inhibition Therapy on The Serum Levels of IL-33 and IL-6 in Patients with Rheumatoid Arthritis

Background Several pro-inflammatory cytokines such as TNF-α, IL-1, IL-6, IL-8 and IL-15 are known to be critical in synovial inflammatory process in RA and successful results have been obtained in RA treatment with targeting pro-inflammatory cytokines including TNF-α, IL-1 and IL-6. Objectives This study sought to investigate the role of IL-33 and IL-6 in RA patients receiving IL-6 receptor inhibition therapy. Methods We analyzed the association of the IL-33 and IL-6 level with disease activity and serologic features in 83 patients with RA. We also measured the serum level of IL-33 and IL-6 before and after the administration of tocilizumab for 24 weeks in 40 patients. Results Serum IL-33 level showed significant correlation with RF (rho =0.660, p<0.001) but did not correlate with DAS28, ESR, hsCRP or RA duration. IL-6 level was significantly correlated with hsCRP (rho =0.482, p<0.001) but not correlate with DAS28, ESR, RF or RA duration. There was no correlation between serum IL-6 and IL-33 levels. Serum IL-33 level significantly decreased after 24 weeks of IL-6 receptor inhibition in patients with RA (p<0.001). When comparing subgroups according to ACR20 response, serum IL-33 levels were significantly decreased after 24 weeks of IL-6 receptor inhibition therapy in ACR20 responders (p<0.001) but not in the non-responders (p=0.084). Baseline IL-33 levels were not significantly different between the two subgroups (p=0.765). Serum IL-6 levels were not significantly changed after 24 weeks of IL-6 receptor inhibition therapy (median 7.1 to 8.9 pg/mL, p=0.503). Changes of IL-6 levels were insignificant both in ACR20 responders and non-responders after 24 weeks of IL-6 receptor inhibition therapy. Baseline IL-6 levels were not different between ACR20 responders and non-responders. Conclusions The use of IL-6 receptor inhibitor decreased the serum level of IL-33 and this effect seems to be led by the responder group. IL-33 could be a useful indicator to monitor the response in IL-6 receptor inhibition therapy. Disclosure of Interest None declared