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Background/Aims: The aim of this study was to investigate long-term post-discontinuation outcomes in patients with rheumatoid arthritis (RA) who had been treated with tumor necrosis factor-α inhibitors (TNF-αi) which was then discontinued.Methods: Sixty Korean patients with RA who participated in a 5-year GO-BEFORE and GO-FORWARD extension trials were included in this retrospective study. Golimumab was deliberately discontinued after the extension study (baseline). Patients were then followed by their rheumatologists. We reviewed their medical records for 2 years (max 28 months) following golimumab discontinuation. Patients were divided into a maintained benefit (MB) group and a loss-of-benefit (LB) group based on treatment pattern after golimumab discontinuation. The LB group included patients whose conventional disease-modifying antirheumatic drug(s) were stepped-up or added/switched (SC) and those who restarted biologic therapy (RB).Results: The mean age of patients at baseline was 56.5 years and 55 (91.7%) were females. At the end of follow-up, 23 (38.3%) patients remained in the MB group. In the LB group, 75.7% and 24.3% were assigned into SC and RB subgroups, respectively. Fifty percent of patients lost MB after 23.3 months. Demographics and clinical variables at baseline were comparable between MB and LB groups except for age, C-reactive protein level, and corticosteroid use. Restarting biologic therapy was associated with swollen joint count (adjusted hazard ratio [HR], 1.90; 95% confidence interval [CI], 1.01 to 3.55) and disease duration (adjusted HR, 1.12; 95% CI, 1.02 to 1.23) at baseline.Conclusions: Treatment strategies after discontinuing TNF-αi are needed to better maintain disease control and quality of life of patients with RA.

Objectives To assess the efficacy of an endothelin receptor antagonist (ERA) and phosphodiesterase type5 inhibitors (PDE5is) for treating SSc-related digital ulcers (DUs). Methods This prospective, multicentre, observational cohort study recruited patients with active SSc-related DUs from 13 medical centres in South Korea. The primary outcome was time to cardinal ulcer (CU) healing. A secondary outcome was time to new DU occurrence. Patients were followed up 4, 8, 12 and 24 weeks after treatment initiation. Results Sixty-three patients were analysed. Their mean age was 49.9 years (s.d. 11.4) and 49 were female. Twenty-eight had limited SSc. Forty-nine patients received ERA, 11 received a PDE5i (9 sildenafil, 1 udenafil and 1 tadalafil) and 3 received other medication. The hazard ratio (HR) for time to CU healing in the ERA group vs the PDE5i group was 0.75 (95% CI 0.35, 1.64; P = 0.47) in an unadjusted model and 0.80 (95% CI 0.36, 1.78; P = 0.59) in a model adjusted for age, sex, use of calcium channel blockers (CCBs), total DU number and initial CU area. The HR for new DU development in the ERA group vs the PDE5i group was 0.39 (95% CI 0.16, 0.93; P = 0.03) in an unadjusted model and 0.32 (95% CI 0.13, 0.81; P = 0.02) in an adjusted model. No patients receiving CCBs developed new DUs at 24 weeks. Conclusion Time to CU healing is comparable for ERA and PDE5i. ERAs are more effective in reducing new DU occurrence than PDE5is. CCBs may be effective as a background medication.

In osteoarthritis (OA), chondrocytes in cartilage undergo phenotypic changes and senescence, restricting cartilage regeneration and favoring disease progression. Although senescence biomarker p16INK4a expression is known to induce aging by halting the cell cycle, therapeutic applications for p16INK4a targeting are limited. Here, we aimed to reduce cartilage damage and alleviate pain using p16INK4a nanoparticles in OA. The p16INK4a expression of human OA chondrocytes and synoviocytes from patients with knee OA was measured and the levels of p16INK4a, tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, and matrix metalloproteinase (MMP) 13 were examined. p16INK4a siRNA was encapsulated into poly (lactic

This study aimed to examine the role of CD70, which is highly expressed on fibroblast-like synoviocytes (FLS), in rheumatoid arthritis (RA) patients. FLS isolated from RA (n = 14) and osteoarthritis (OA, n = 4) patients were stimulated with recombinant interleukin-17 (IL-17; 5 ng/mL) and tumor necrosis factor alpha (TNF-α; 5 ng/mL) for 24 h. Expression of CD70, CD27/soluble CD27 (sCD27), and hypoxia-inducible factor-2 alpha (HIF-2α) was analyzed by RT-qPCR, flow cytometry, and ELISA assays, respectively. Reactive oxygen species (ROS) expression and cell migration were also examined. The HIF-2α inhibitor PT-2385 and CD70 inhibitor BU69 were used to specifically suppress these pathways. Stimulation with IL-17 and TNF-α significantly induced CD70 expression in RA FLS. Although the synovial fluids from patients with RA contained high levels of sCD27, surface expression of CD27, a ligand of CD70, was rarely detected in RA FLS. Cytokine-induced CD70 expression was significantly decreased following antioxidant treatment. Following HIF-2α inhibition, RA FLS had decreased expression of CD70 and ROS levels. Migration of RA FLS was also inhibited by inhibition of CD70 or HIF-2α. The surface expression of CD70 is regulated by HIF-2α and ROS levels and is a key contributor to cytokine-enhanced migration in RA FLS.

The essential micronutrient zinc regulates immune responses by affecting signaling pathways. In activated monocytes and macrophages, signaling networks mediate the metabolic reprogramming that meets the demands of participation in immune responses. Here, we demonstrated that cytoplasmic, bioavailable zinc was essential for promoting IL-1β production in activated human monocytes and macrophages downstream of glycolysis induced by the kinase-containing multiprotein complex mTORC1. The concentration of cytoplasmic zinc was determined by that of extracellular zinc, which was brought into cells through the zinc-specific importer Zip8. The abundance of Zip8 was increased in monocytes from patients with rheumatoid arthritis (RA), as well as in LPS-stimulated monocytes and macrophages from healthy individuals. The mTORC1-mediated phosphorylation of S6 kinase (S6K) was enhanced by zinc-mediated inhibition of PP2A, a phosphatase that targets S6K. As a result, IL-1β production was increased due to the activation of mTORC1-induced glycolysis. In monocytes of patients with RA, the expression of

The production and oxidation mechanism of reactive oxygen species (ROS) are out of balance in rheumatoid arthritis (RA). However, the correlation between ROS and T cell subsets in RA remains unclear. Peripheral blood mononuclear cells (PBMCs) from patients with RA (n = 40) and healthy controls (n = 10) were isolated from whole blood samples. Synovial tissues (n = 3) and synovial fluid (n = 10) were obtained from patients with RA. The repartition of T cell subsets and expression of ROS and cytokines were examined according to RA severity. Fibroblast-like synoviocytes (FLSs) from patients with RA were stimulated with PBMCs and the expression of inflammation-related molecules were measured by RT-PCR and cytokine array. Regulatory T cells from patients with moderate (5.1 &gt, DAS28 ≥ 3.2) RA showed the highest expression of mitochondrial ROS among the groups based on disease severity. Although ROS levels steadily increased with RA severity, there was a slight decline in severe RA (DAS28 ≥ 5.1) compared with moderate RA. The expression of inflammatory cytokines in RA FLSs were significantly inhibited when FLSs were co-cultured with PBMCs treated with ROS inhibitor. These findings provide a novel approach to suppress inflammatory response of FLSs through ROS regulation in PBMCs.

There is growing evidence that apurinic/apyrimidinic endonuclease 1/redox factor-1 (APE1/Ref-1) regulates inflammatory responses. Rheumatoid arthritis (RA) is an autoimmune disease, which is characterized with synovitis and joint destruction. Therefore, this study was planned to investigate the relationship between APE1/Ref-1 and RA. Serum and synovial fluid (SF) were collected from 46 patients with RA, 45 patients with osteoarthritis (OA), and 30 healthy control (HC) patients. The concentration of APE1/Ref-1 in serum or SF was measured using the sandwich enzyme-linked immunosorbent assay (ELISA). The disease activity in RA patients was measured using the 28-joint disease activity score (DAS28). The serum APE1/Ref-1 levels in RA patients were significantly increased compared to HC and OA patients (0.44 ± 0.39 ng/mL for RA group vs. 0.19 ± 0.14 ng/mL for HC group, p &lt, 0.05 and vs. 0.19 ± 0.11 ng/mL for OA group, p &lt, 0.05). Likewise, the APE1/Ref-1 levels of SF in RA patients were also significantly increased compared to OA patients (0.68 ± 0.30 ng/mL for RA group vs. 0.31 ± 0.12 ng/mL for OA group, p &lt, 0.001). The APE1/Ref-1 concentration in SF of RA patients was positively correlated with DAS28. Thus, APE1/Ref-1 may reflect the joint inflammation and be associated with disease activity in RA.

Objectives: This study aims to investigate the role of cluster of differentiation 14 (CD14) expressed monocytes and soluble CD14-mediated pathway in the synovial inflammation of knee osteoarthritis (OA). Patients and methods: Between May 2012 and July 2013, a total of 35 patients with knee OA (9 males, 26 females; mean age: 66.3±8.8 years; range, 52 to 79 years) were included in this cross-sectional study. Synovial fluid was obtained from knee joints of 35 OA patients. The CD14+ monocytes from synovial fluid mononuclear cells (SFMCs) were isolated using the MACS. The fibroblast-like synoviocytes (FLSs) isolated from knee joint tissue were incubated with recombinant CD14 and lipopolysaccharide (LPS) for 24 h. Cytokine profiling was performed with the Luminex® Performance Assay or magnetic bead panel kit. The expression of CD14 and CD16 was analyzed by immunohistochemistry and flow cytometry. Results: The concentration of sCD14 in synovial fluid was correlated with the interleukin-6 (IL-6) level (n=35) (ρ=0.654, p+ monocytes isolated from SFMC (n=15) showed a correlation between sCD14 and IL-6 (ρ=0.784, p=0.001), along with complement component 3 (ρ=0.756, p=0.010), IL-1b (ρ=0.652, p=0.012), and tumor necrosis factor-alpha (ρ=0.806, p=0.001). Following recombinant CD14 and LPS treatment, OA FLS synergistically enhanced the secretion of IL-6, IL-8, and matrix metalloproteinase 3 (n=3, p+ monocytes were significantly elevated in recurred synovial fluid compared to those in initial synovial fluid (p=0.043). When monocyte subsets were analyzed in SFMC (n=26), CD14+CD16+monocytes were abundant (p=0.019) and had higher toll-like receptor 4 expression than CD14+CD16- (p Conclusion: Our study results suggest that CD14+ monocytes and the sCD14-mediated pathway play an important role in OA aggravation through inflammatory cytokine secretion.

Objectives This study aims to examine the possible associations of mitochondrial single nucleotide polymorphisms (SNPs) and Behcet's disease (BD) in a larger patient group. Patients and methods Whole blood or buffy coat was collected from 98 BD patients (31 males, 67 females; mean age 48±2.8 years; range 20 to 60 years) from four university hospitals located in the Chung-Cheong district of the Republic of Korea, and 196 age- and sex-matched healthy controls (HCs) (62 males, 134 females; mean age 46.91±12.90 years; range 20 to 68 years) from Konyang University Hospital. Twenty targeted mitochondrial deoxyribonucleic acids (DNAs) were genotyped and compared using the revised Cambridge Reference Sequence. Chi square and Fisher's exact tests were used to analyze association of mitochondrial DNA SNPs with BD susceptibility and its clinical characteristics. Results There were no differences for m.248A>G, m.304C>A, m.709G>A, m.3010G>A, m.3970C>T, m.4883C>T, m.5178C>A, m.6392T>C, m.6962G>A, m.10310G>A, m.10609T>C, m.12406G>A, m.12882C>T, m.13928G>C, m.14668C>T, m.16129G>A, and m16304T> between patient and HC groups. However, m.16182A>C and m.16183A>C were more frequently observed in the patient group than the HC group (22 [22.4%] vs. 24 [12.2%], p=0.061 and 32 [32.7%] vs. 42 [21.4%], p=0.092) but without statistical significance. m.4883C>T and m.5178C>A were associated with posterior location of oral ulcers (p=0.025 for each) and m.16183A>C was associated with deep oral ulcers (p=0.001), while m.16189T>C was associated with deep oral ulcers and thrombosis (p=0.042, 0.048, respectively). Conclusion m.16182A>C and m.16183A>C may be associated with BD in the Korean population.

Background Patient-centered management is becoming increasingly important in gout, but there are limited studies exploring patients' perspectives and preferences. We aimed to investigate patients' perspectives and preferences regarding gout and gout management, and their impacts on adherence to urate lowering therapy (ULT). Methods A paper-based survey was performed in patients with gout seen at the rheumatology outpatient clinics of 16 tertiary hospitals. The survey included questions regarding demographics, comorbidities, gout attacks, current treatment and adherence, and patients' perspectives and preferences regarding gout and gout management. Multivariate regression analysis was performed to determine the factors associated with ULT adherence. Results Of 809 surveyed patients with gout, 755 (94.5%) were using ULT. Among those using ULT, 89.1% had ≥ 80% adherence to ULT. Majority of the patients knew management strategies to some extent (94.8%), perceived gout as a life-long disease (91.2%), and were making efforts toward practicing at least one lifestyle modification (89.2%). Most patients (71.9%) obtained information about gout management during their clinic visits. Approximately half of the patients (53.6%) preferred managing their disease with both ULT and lifestyle modification, 28.4% preferred ULT only, and 17.4% preferred lifestyle modification only. Adherence was better in patients with older age (odds ratio [OR], 1.03), those with better knowledge of gout management strategies (OR, 3.56), and those who had preference for ULT (OR, 2.07). Conclusion Patients' perspectives and management preferences had high impacts on adherence to ULT in gout. Consideration of patients' perspectives and preferences is important for achieving the desired clinical outcome in gout., Graphical Abstract

Rheumatoid arthritis (RA) is a common autoimmune disease characterized by immune cell infiltration of the synovium, leading to the loss of cartilage, bone, and joint function. Although regulatory T (Treg) cells are thought to modulate the initiation and progression of RA, a consensus has yet to be reached regarding the function and composition of Treg cells in RA patients. To address these discrepancies, we analyzed not only the total Treg frequency but also that of Treg subpopulations in the peripheral blood of RA patients and healthy controls by flow cytometry. We found that the total Treg population was not significantly different between RA and control subjects. However, the effector Treg cell subgroup, defined as CD45RA−CD25hi, showed markedly decreased frequency in RA patients. In addition, the total Treg population from RA patients showed a significant decline in the expression of CD25. Both the naïve and effector Treg subgroups also showed marked reduction of CD25 expression in RA patients compared to controls. These data suggest that the decreased frequency of effector Treg cells and overall reduction of CD25 expression in Treg cells in the peripheral blood may be evidence of altered Treg homeostasis associated with RA pathogenesis.

Background Reactive oxygen species (ROS) regulate the migration and invasion of fibroblast-like synoviocytes (FLS), which are key effector cells in rheumatoid arthritis (RA) pathogenesis. Nicotinamide adenine dinucleotide phosphate oxidase 4 (NOX4) induces ROS generation and, consequently, enhances cell migration. Despite the important interrelationship between RA, FLS, and ROS, the effect of NOX4 on RA pathogenesis remains unclear. Methods FLS isolated from RA (n = 5) and osteoarthritis (OA, n = 5) patients were stimulated with recombinant interleukin 17 (IL-17; 10 ng/ml) and tumor necrosis factor alpha (TNF-α; 10 ng/ml) for 1 h. Cell migration, invasion, adhesion molecule expression, vascular endothelial growth factor (VEGF) secretion, and ROS expression were examined. The mRNA and protein levels of NOX4 were analyzed by RT-qPCR and western blotting, respectively. The NOX4 inhibitor GLX351322 and NOX4 siRNA were used to inhibit NOX4 to probe the effect of NOX4 on these cellular processes. Results Migration of RA FLS was increased 2.48-fold after stimulation with IL-17 and TNF-α, while no difference was observed for OA FLS. ROS expression increased in parallel with invasiveness of FLS following cytokine stimulation. When the expression of NOX was examined, NOX4 was significantly increased by 9.73-fold in RA FLS compared to unstimulated FLS. Following NOX4 inhibition, cytokine-induced vascular cell adhesion molecule 1 (VCAM1), VEGF, and migration and invasion capacity of RA FLS were markedly decreased to unstimulated levels. Conclusion NOX4 is a key contributor to cytokine-enhanced migration and invasion via modulation of ROS, VCAM1, and VEGF in RA FLS.

Objective To evaluate fenebrutinib, an oral and highly selective noncovalent inhibitor of Bruton's tyrosine kinase (BTK), in patients with active rheumatoid arthritis (RA). Methods Patients with RA and an inadequate response to methotrexate (MTX) (cohort 1; n = 480) were randomized to receive fenebrutinib (50 mg once daily, 150 mg once daily, or 200 mg twice daily), adalimumab (40 mg every other week), or placebo. Patients with RA and an inadequate response to tumor necrosis factor inhibitors (cohort 2; n = 98) received fenebrutinib (200 mg twice daily) or placebo. Both cohorts continued MTX therapy. Results In cohort 1, the percentages of patients in whom American College of Rheumatology 50% improvement criteria (ACR50) was achieved at week 12 were similar in the fenebrutinib 50 mg once daily and placebo groups, and were higher in the fenebrutinib 150 mg once daily group (28%) and 200 mg twice daily group (35%) than in the placebo group (15%) (P = 0.016 and P = 0.0003, respectively). Fenebrutinib 200 mg twice daily and adalimumab (36%) were comparable (P = 0.81). In cohort 2, ACR50 was achieved in more patients receiving fenebrutinib 200 mg twice daily (25%) than placebo (12%) (P = 0.072). The most common adverse events in the fenebrutinib groups included nausea, headache, anemia, and upper respiratory tract infections. Fenebrutinib had significant effects on myeloid and B cell biomarkers (CCL4 and rheumatoid factor). Fenebrutinib and adalimumab caused overlapping as well as distinct changes in B cell and myeloid biomarkers. Conclusion Fenebrutinib demonstrates efficacy comparable to adalimumab in patients with an inadequate response to MTX, and safety consistent with existing immunomodulatory therapies for RA. These data support targeting both B and myeloid cells via this novel mechanism for potential efficacy in the treatment of RA.

Summary Background Ixekizumab, a high-affinity interleukin-17A (IL-17A) monoclonal antibody, has previously shown efficacy in radiographic axial spondyloarthritis (also known as ankylosing spondylitis). We aimed to evaluate the efficacy and safety of ixekizumab, an IL-17 inhibitor, in non-radiographic axial spondyloarthritis. Here, we report the primary results of COAST-X. Methods COAST-X was a 52-week, randomised, double-blind, placebo-controlled, parallel-group study done at 107 sites in 15 countries in Europe, Asia, North America, and South America. Eligible participants were adults (aged ≥18 years) with active axial spondyloarthritis without definite radiographic sacroiliitis (non-radiographic axial spondyloarthritis), objective signs of inflammation (via MRI or C-reactive protein), and an inadequate response or intolerance to non-steroidal anti-inflammatory drugs (NSAIDs). Patients were randomly assigned (1:1:1) to receive subcutaneous 80 mg ixekizumab every 4 weeks (Q4W) or every 2 weeks (Q2W), or placebo. Changing background medications or switching to open-label ixekizumab Q2W, or both, was allowed after week 16 at investigator discretion. Primary endpoints were Assessment of SpondyloArthritis international Society-40 (ASAS40) response (defined as an improvement of 40% or more and an absolute improvement from baseline of 2 units or more [range 0–10] in at least three of the four domains [patient global, spinal pain, function, and inflammation] without any worsening in the remaining one domain) at weeks 16 and 52. Patients who switched to open-label ixekizumab were imputed as non-responders in logistic regression analysis. This trial is registered with ClinicalTrials.gov , number NCT02757352 . Findings Between Aug 2, 2016, and Jan 29, 2018, 303 patients were enrolled (105 to placebo, 96 to ixekizumab Q4W, and 102 to ixekizumab Q2W). Both primary endpoints were met: ASAS40 at week 16 (ixekizumab Q4W: 34 [35%] of 96, p=0·0094 vs placebo; ixekizumab Q2W: 41 [40%] of 102, p=0·0016; placebo: 20 [19%] of 105) and ASAS40 at week 52 (ixekizumab Q4W: 29 [30%] of 96, p=0·0045; ixekizumab Q2W: 32 [31%] of 102, p=0·0037; placebo: 14 [13%] of 105). 60 (57%) of 104 patients in the placebo group, 63 (66%) of 96 in the ixekizumab Q4W group, and 79 (77%) of 102 in the ixekizumab Q2W group had at least one treatment-emergent adverse event. The most common treatment-emergent adverse events in the ixekizumab groups were nasopharyngitis and injection site reaction. Of the treatment-emergent adverse events of special interest, there was one case of serious infection in the ixekizumab Q4W group. The frequency of serious adverse events was low (four [1%] of 302) and similar across the three groups. There were no malignancies or deaths. No new safety signals were identified. Interpretation Ixekizumab was superior to placebo for improving signs and symptoms in patients with non-radiographic axial spondyloarthritis at weeks 16 and 52. Reports of adverse events were similar to those of previous ixekizumab studies. Ixekizumab offers a potential therapeutic option for patients with non-radiographic axial spondyloarthritis who had an inadequate response or were intolerant to NSAID therapy. Funding Eli Lilly and Company.

Chromosomes of Korean hexaploid wheat were investigated to compare the chromosomal karyotype for cytogenetic diversity. Chromosomal karyotyping was done with in situ hybridization using two types of simple sequence repeats (SSR)s, (AAG)5 and (AAC)5 labeled with tetramethyl-rhodamine-5-dUTP and fluorescein-12-dUPT as a fluorescence, respectively. The two SSRs as cytogenetic markers revealed that the cytogenetic characteristics of the wheat chromosomes were remarkably a B genome. In this study, the chromosomal karyotype of Keumkang, a Korean hexaploid wheat cultivar, was the A, B, and D genomes used as a cytogenetic reference. The expressed signals from the two SSRs showed a difference in the chromosomal karyotype of chromosome 1B among the Korean hexaploid wheat. The distribution pattern and the degree of condensation for the (AAG)5 and (AAC)5 signals on the short arm of chromosome 1B were different in the Korean hexaploid wheat shown in descending order: Keumkang > Joeun > Johan > Olgeuru. Olgeuru had a lower level of distribution and condensation for the two SSRs signals compared to the other Korean hexaploid wheat. In the A genome, chromosome 7A showed an unbalanced expression of the (AAG)5 signal on the distal region of the short and long arms in several Korean hexaploid wheat while Joeun, a Korean hexaploid wheat, showed a definite (AAG)5 signal on the distal region of each arm of chromosome 7A. Among the Korean hexaploid wheat, Shinmichal1, a Korean hexaploid waxy wheat, had a chromosome with a unique expression pattern for (AAG)5 and (AAC)5 compared the other Korean hexaploid wheat. Those cytogenetic differences identified in this study are useful as an indicator to improve the cytogenetic diversity in the Korean wheat breeding program.

Objectives This study investigated the expression of proviral-integration site for Moloney murine leukaemia virus (PIM) -1 kinase in RA synovium and RA fibroblast-like synoviocytes (FLSs) along with its impact on RA-FLS aggressiveness. Methods The expression of PIM kinases was assessed in synovial tissues by immunohistochemistry and double IF. After PIM-1 inhibition using either small-interfering RNA or the chemical inhibitor AZD1208, we performed proliferation and migration assays and measured the levels of MMPs and IL-6 released from RA-FLSs under stimulation with proinflammatory cytokines (TNF-α, S100A4 and IL-6/soluble IL-6 receptor). Additionally, PIM-1-associated downstream signalling pathways were analysed by immunoblotting. Results Three isoforms of PIM kinases were immunodetected in the synovial tissues from patients with RA or OA. Specifically, PIM-1 and PIM-3 were upregulated in RA synovium and PIM-1 was expressed in T cells, macrophages and FLSs. Additionally, upon stimulation of RA-FLSs with TNF-α, S100A4 and IL-6/sIL-6R, PIM-1 and PIM-3, but not PIM-2, were significantly inducible. Moreover, PIM-1 knockdown or AZD1208 treatment significantly suppressed basal or cytokine-induced proliferation and migration of RA-FLS and the secretion of MMPs from stimulated RA-FLSs. PIM-1 knockdown significantly affected the phosphorylation levels of extracellular signal-regulated kinase and cAMP responsive element binding protein in RA-FLSs. Conclusion PIM-1 was upregulated in RA synovial tissues and RA-FLSs and its inhibition significantly reduced the proliferation, migration and MMP production of RA-FLSs in vitro. These findings suggest PIM-1 as a novel regulator of the aggressive and invasive behaviour of RA-FLSs and indicate its potential as a target for RA treatment.

PURPOSE Although cardiac manifestation of Behcet disease (BD) has been described in sporadic reports, its timely diagnosis remains difficult. The objective of this study was to describe early cardiac manifestations of BD. We also performed a comprehensive classification of systemic BD activity and compared their cardiac manifestations. METHODS A prospective screening using speckle tracking echocardiography was performed in 85 patients with BD who had no history of heart disease. After excluding subjects with left ventricular (LV) ejection fraction (LVEF)

This study assessed the relationships among the risk factors for and components of metabolic syndrome (MetS) and health-related quality of life (HRQOL) in a hypothesized causal model using structural equation modeling (SEM) in patients with systemic lupus erythematosus (SLE). Of the 505 SLE patients enrolled in the Korean Lupus Network (KORNET registry), 244 had sufficient data to assess the components of MetS at enrollment. Education level, monthly income, corticosteroid dose, Systemic Lupus Erythematosus Disease Activity Index, Physicians’ Global Assessment, Beck Depression Inventory, MetS components, and the Short Form-36 at the time of cohort entry were determined. SEM was used to test the causal relationship based on the Analysis of Moment Structure. The average age of the 244 patients was 40.7 ± 11.8 years. The SEM results supported the good fit of the model (χ 2 = 71.629, p = 0.078, RMSEA 0.034, CFI 0.972). The final model showed a direct negative effect of higher socioeconomic status and a positive indirect effect of higher disease activity on MetS, the latter through corticosteroid dose. MetS did not directly impact HRQOL but had an indirect negative impact on it, through depression. In our causal model, MetS risk factors were related to MetS components. The latter had a negative indirect impact on HRQOL, through depression. Clinicians should consider socioeconomic status and medication and seek to modify disease activity, MetS, and depression to improve the HRQOL of SLE patients.

Objectives The objectives of this study were to evaluate the long-term effect of anti-platelet treatment on the radiological progression of collagen-induced arthritis in rats. Methods Female Lewis rats with collagen-induced arthritis were divided into three experimental groups: saline, aspirin monotherapy (n = 12), and aspirin-clopidogrel dual therapy (n = 12). Drugs were administered daily and continued up to 70 days after the induction of arthritis. The clinical arthritis index (weight, morphology score, and paw thickness) and radiological scores were evaluated. Results The clinical arthritis index peaked on day 20, while the radiological scores peaked on day 35. No intergroup difference was observed in the clinical arthritis index throughout the experiment. The aspirin-clopidogrel dual therapy group had a significantly higher mean radiological score than the other groups (p = 0.045) on day 35. Further treatments resulted in significantly improved radiological findings in the aspirin monotherapy and aspirin-clopidogrel dual therapy groups on day 70 but no significant improvement in the saline group. Conclusion Anti-platelet agent treatment improved radiological findings on day 70. These observations emphasize the importance of a future long-term study of the effects of anti-platelet agent treatment on arthritis.

Background Rheumatoid arthritis (RA) is characterized by inflammation and cellular proliferation in the synovium. Activated lymphocytes and proinflammatory molecules are important in the pathogenesis of RA. CD70 belongs to the tumor necrosis factor (TNF) ligand superfamily and is typically present on activated B and T lymphocytes, natural killer cells and mature dendritic cells. CD70 expressing CD4+ T cells are enriched in the peripheral blood and synovial fluid of patients with RA and promote autoimmunity via co-stimulatory CD70-CD27 interaction. CD70 expression is associated with aggressive phenotype of cancer cells and it is mediated by hypoxia inducible factor 2α (HIF-2α). Objectives In this study, we examined the presence of CD70 on the surface of fibroblast-like synoviocyte (FLS) of patients with RA (RA-FLS) and investigate the role of CD70 in the pathogenesis of RA associated with HIF-2α. Methods RA FLS were obtained from 7 patients with RA who were undergone operation like total knee replacement or synovectomy. All patients were fulfilled the 2010 ACR-EULAR classification criteria for RA. CD70 and HIF-2α messenger ribonucleic acid (mRNA) were analyzed in RA-FLS by quantitative polymerase chain reaction (qPCR). CD70 and CD27 on the surface of RA-FLS were stained by PE-Anti CD70 antibody and PerCP-Cy5.5-CD27 antibody respectively and evaluated by flow cytometry. Same experiments were performed after treatment with interleukin (IL)-17, TNF-α and HIF-2α blocking antibody (Anti HIF-2α antibody). Results CD70 and HIF-2α mRNA in the RA-FLS were elevated after treatment with IL-17 and TNF-α (Figure 1, 2). The level of CD70 expression on the surface of RA-FLS was elevated after stimulation with IL-17 and TNF-α (Figure 3). And it was lowered after treatment with HIF-2α blocking antibody as dose dependent pattern (Figure 3). CD27 wasn’t present on the surface of RA-FLS (Figure 4). Conclusion We identified the expression of CD70 on the surface of RA-FLS. And in inflammatory conditions like stimuation with IL-17 and TNF-α, both CD70 and HIF-2α mRNA were increased. The level of CD70 on the surface of RA-FLS also elevated by treatment with IL-17 and TNF-α. The result of decreased level of CD70 after treatment with anti HIF-2α antibody suggest that CD70 expression on the surface of RA-FLS is associated with HIF-2α. From these results, we expect that CD70-targeted therapy associated with HIF-2α may be effective for treatment with RA. References [1] Han, B. K., Olsen, N. J., & Bottaro, A. (2016). The CD27-CD70 pathway and pathogenesis of autoimmune disease. Semin Arthritis Rheum, 45(4), 496-501. [2] Kitajima, S., Lee, K. L., Fujioka, M., Sun, W., You, J., Chia, G. S. Poellinger, L.(2018). Hypoxia-inducible factor-2 alpha up-regulates CD70 under hypoxia and enhances anchorage-independent growth and aggressiveness in cancer cells. Oncotarget, 9(27), 19123-19135. [3] Lee, W. W., Yang, Z. Z., Li, G., Weyand, C. M., & Goronzy, J. J. (2007). Unchecked CD70 expression on T cells lowers threshold for T cell activation in rheumatoid arthritis. J Immunol, 179(4), 2609-2615. [4] Oflazoglu, E., Boursalian, T. E., Zeng, W., Edwards, A. C., Duniho, S., McEarchern, J. A. Grewal, I. S. (2009). Blocking of CD27-CD70 pathway by anti-CD70 antibody ameliorates joint disease in murine collagen-induced arthritis. J Immunol, 183(6), 3770-3777. [5] Park, J. K., Han, B. K., Park, J. A., Woo, Y. J., Kim, S. Y., Lee, E. Y., Song, Y. W. (2014). CD70-expressing CD4 T cells produce IFN-gamma and IL-17 in rheumatoid arthritis. Rheumatology (Oxford), 53(10), 1896-1900. [6] Ruf, M., Moch, H., & Schraml, P. (2015). Interaction of tumor cells with infiltrating lymphocytes via CD70 and CD27 in clear cell renal cell carcinoma. OncoImmunology, 4(12), e1049805. Disclosure of Interests None declared

Background: There was some reports of seasonal association with myopsitis onset. With the discovery of new myositis-specific autoantibodies (MSA), detailed grouping of idiopathic inflammatory myopathies was possible. Therefore, we evaluated the seasonal patterns in the onset of idiopathic inflammatory myositis (IIM) with MSA in Korea. Objectives: To evaluate the correlation between MSA and seasonal patterns of IIM in Korea. Methods: A total of 90 patients who met the criteria for probable or definite PM or DM and for whom data were collected from 7 referal centers in korea. 16 myositis-specific autoantibodies were detected by immunoblot with patient’s serum. Statistical analyses were performed using a Poisson model that assessed associations of sex, MSA, and month of onset of symptoms or month of diagnosis. Results: There were no significant seasonal patterns of disease onset in total IIM patients. Among MSAs, anti-synthetase (n=18), anti-transcriptional intermediary factor 1 γ (TIF1γ) (n=13), anti-melanoma differentiation-aasocianted protein 5 (anti-MDA5) (n=12), and anti–signal recognition particle (SRP) (n=12) were analyzed. Among 55 patients with dermatomyositis, patients with anti-MDA5 showed a significant peak in winter (n=12, P=0.05). This seasonal association was significant in women (n =10; P=0.045). Patients with anti–synthetase, anti-TIF1 γ or anti-SRP antibodies did not have a significant seasonal onset patterns. There were no significant seasonal patterns in patients without myositis-specific autoantibodies. Conclusion: Patients with anti-MDA5 showed a seasonality of myositis onset, in winter. Disclosure of interests: None declared

Background SinceBohan and Peter first described their diagnostic criteria foridiopathic inflammatorymyopathies (IIM) in 1975, new discoveries suchas myositis-specific and myositis-associated autoantibodies (Abs) have been made. Objectives To investigate correlations between specificmyositis Abs and their frequenciesand clinical associationsacross different IIM groups, collectively demonstrating theutility of the new clinicoserologic classification in Koreanadult patients with IIM. Methods We conducted a multicenter cohort study including 67adult patients (age≥18 years) who have been diagnosed as IIM by ENMC criteria.Immunoblot assay with Euroline strip(EUROIMMUN, Germany)was performed using the sera of definite deramatomyositis (DM, n=36), definite polymyositis (PM, n=25), amyopathic DM (n=4), DM sine dermatitis (n=1), and immune mediated necrotizing myopathy (IMNM, n=1). Patients were classifiedbased on three classifications: 1) novel clinicoserologic classification suggested by Troyanov et al. in 2017. 2) 2017 EULAR/ACR classification criteria. 3) 2004 European neuromuscular center (ENMC) criteria.Associations ofmyositis Absand clinical subsets of IIM were investigated. Results The distribution of the various IIM differed strikingly from those using the 3 classifications (Fig1). According to the 2004 ENMC classification and 2017 EULAR/ACR classification criteria, DM and PM was the most and the second frequent entities (DM: 55.2%, 56.7%; PM: 35.8%, 37.3%). But, using the new clinicoserologic classification,overlapmyositis(OM) is the major type of IIM and the frequency of PM is significantly decreased.Anti-ARS Abs specificity included anti-Jo-1(16.4%), -OJ(4.6%), -EJ(6.2%) -PL-7(3.1%), and -PL-12(4.6%). Interstitial lung disease was closely associated with anti-MDA5,and anti-ARS Abs, while DM-specific skin lesion was frequently observed in patients with anti-TIF1γ and anti-ARS Abs. Sevenpatients with cancer-associated DM were identified. They were positive for anti-TIFγ (5/7) and anti- SRP(3/7) (table 1). Conclusion Novel classification based on distinctive features and new myositis Absreflects the clinical phenotype of IIM better. Establishment of a system routinely available to screen myositis Abs is needed. Thiswill be beneficial to providemore precise diagnosis and proper management for patients with IIM. Reference: [1] Jean-Luc Senecal, Jean-Pierre Raynauld, Yves Troyanov. A New Classification of Adult Autoimmune Myositis. Arthritis Rheum2017;69:878-884. Disclosure of Interests None declared

Background: Myositis-specific and myositis-associated autoantibodies (MSA and MAA) have been used for more detailed classification and prediction of clinical course of idiopathic inflammatory myopathies (IIM). Interstitial lung disease (ILD) is known to be complicated with approximately 50% of IIM patients, having poor prognosis. Objectives: The aim of this study is to investigate clinical features of ILD in Korean adult patients with IIM according to the autoantibody profile including MSA and MAA. Methods: We conducted a multicenter cohort study including 108 adult patients (age≥18 years) who have been diagnosed as IIM by ENMC criteria MSA and MAA were screened with Immunoblot assay using Euroline strip (EUROIMMUN, Germany). Clinical information including the time of developing ILD and imaging findings of chest CT was reviewed using by medical record. Results: ILD was found in about half (52.8%, 57 of 108) of enrolled patients and closely associated with the presence of anti-ARS Abs, anti-MDA5 and anti-Ro52. Among 43 patients, Anti-ARS Abs were most common (46.5%) and included anti-Jo-1 (23.3%), -OJ (2.3%), -EJ (9.3%), -PL7 (14%), and -PL12 (7%) (Figure 1). To analyze the correlation of autoantibodies with pattern of ILD onset, we stratified the 43 patients into three groups: into ILD-preceding (7/43, 16.3%), simultaneous (31/43, 72.1%) and myopathy-preceding (6/43, 14%). In the ILD-preceding group, anti-ARS antibodies were frequent. ILD preceded myopathy by 19.5 months on average. The imaging pattern of ILD in the ILD-preceding group were Cryptogenic Organizing Pneumonia (COP) 6, Nonspecific Interstitial Pneumonia (NSIP) 1 and no Usual interstitial pneumonia (UIP). A half of patient in ILD-preceding group had Raynaud`s phenomenon, and one patient had mechanic hand. On the other hand, in myopathy-preceding group, 30% of patients (2/6) were UIP and a half of patients had pulmonary function which required treatment (DLCO Conclusion: In our study, ILDs were accompanied by over half of the cases of IIM and most of them had autoantibodies. There are characteristic clinical features by onset time of ILD, so paying attention to them will help early diagnosis and prognosis prediction of ILDs. Reference [1] Yoshifuji H. Clin Med Insights Circ Respir Pulm Med. 2016 Apr 3;9 (Suppl 1):141-6. Disclosure of Interests: None declared

Background T cells play a regulatory role in rheumatoid arthritis (RA) through inducing the homeostasis maintenance and self-tolerance [1]. Specially, the production and the oxidation mechanism of reactive oxygen species (ROS) were out of balance. Objectives The aim of the study was to compare ROS productions in T cell subset, which are helper T (TH) cell, cytotoxic T (TC) cell, T helper 17 (TH17) cell and regulatory T (Treg) cell in peripheral blood mononuclear cells (PBMC) of RA patients with RA activity. Methods Blood samples were collected from 30 RA patients and 10 healthy adult volunteers under IRB approval. RA activity was divided according to clinical parameter DAS28 [2]. PBMC cells were obtained from the whole blood using lymphocyte separation medium density gradient centrifugation. For separation between the live and dead cell populations, PBMC was stained with Live/Dead stain dye. After PBS washing, cells were incubated with antibodies for CD3, CD4, CD8, and CD25. Following fixation and permeabilization, and further stained with antibodies for FoxP3 and IL-17A. For ROS staining, CellRox and MitoSox were used. Results The frequency of TH cell was increased and that of TC cell was decreased in the peripheral blood of RA patients. TH17 and Treg cell population were significantly increased more than about 2-3 folds in active and inactive RA than healthy control. When the whole of cellular ROS production was measured, only Treg cell population was significantly increased in RA than control. Although ROS level was steadily increased with RA activity, there was a slight decline in severe RA compared to moderate and low RA. This difference is lager in mitochondrial specific ROS than total cellular ROS. The mitochondrial complex inhibitor reduced Treg cell frequency in PBMC from RA patients. Conclusion Treg is the most sensitive to ROS production among T cell subsets in RA. These findings provide a novel approach to regulate Treg function in RA through mitochondrial metabolism related ROS production. References [1] Szekanecz, Z., et al., New insights in synovial angiogenesis. Joint Bone Spine, 2010. 77(1): p.13-9. [2] Prevoo, M.L., et al., Modified disease activity scores that include twenty-eight-joint counts. Development and validation in a prospective longitudinal study of patients with rheumatoid arthritis. Arthritis Rheum, 1995. 38(1): p.44-8. Disclosure of Interests None declared

Purpose To investigate correlations between myositis-specific autoantibodies (MSA) or myositis-associated antibodies (MAA) and clinical features, thereby demonstrating the utility of clinicoserologic classification in idiopathic inflammatory myopathies (IIM) patients. Materials and methods We conducted a multicenter study of 108 adult patients (age ≥18 years) who were diagnosed with IIM by Peter and Bohan criteria or 2004 European Neuromuscular Centre (ENMC) criteria. Clinical data were obtained by medical record review. Immunoblot assay with Euroline strip (EUROIMMUN, Germany) was performed using the sera of dermatomyositis (DM, n=56), polymyositis (PM, n=45), amyopathic DM (n=5), DM sine dermatitis (n=1), and immune mediated necrotizing myopathy (n=1) patients. Patients were classified based on two classifications: 2017 EULAR/ACR and novel clinicoserologic classification. Results According to 2017 EULAR/ACR criteria, DM and PM were the most and the second most frequent entities. Overlap myositis was the major entity of IIM, and the frequency of PM was significantly lower when applying clinicoserologic classification criteria. Sixty-nine (63.9%) patients had one or more MSA, and 61 (56.5%) patients had one or more MAA. Interstitial lung disease was closely associated with anti-MDA5 and anti-ARS, and DM-specific skin lesions were frequently observed in patients with anti-TIF1γ, anti-SRP, and anti-MDA5. Conclusion The clinicoserologic criteria based on MSA/MAA positivity could reflect more precise clinical features of IIM. Establishment of a laboratory system routinely available to screen for MSA/MAA status will be beneficial to provide precise diagnosis and proper management of IIM patients.

BACKGROUND Pulmonary arterial hypertension (PAH) is a major cause of morbidity and mortality among patients with systemic sclerosis (SSc). Early detection and prompt treatment of PAH associated with SSc (SSc-PAH) result in better prognosis. We conducted echocardiographic study to presume the prevalence of PAH in Korean adult SSc patients and to diagnose SSc-PAH in their early stages with right heart catheterization (RHC). METHODS We performed free of charge echocardiographic study including 37 adult SSc patients at the Chungnam National University Hospital. The possibility of PAH is determined by the estimation of pulmonary arterial pressure by peak tricuspid regurgitation velocity of > 3.0 m/s. Patients with possible PAH were recommended to undergo RHC to confirm the diagnosis. RESULTS In 37 patients, 8 patients were suspected with PAH. Among them, 6 patients agreed to be examined with RHC, and 4 were confirmed with PAH. The prevalence of possible PAH was 21.6% (8 of 37 patients), and that of confirmed PAH was 10.8% (4 of 37 patients). Four patients who were confirmed with SSc-PAH through RHC have been treated with specific pulmonary vasodilators and maintained stable. CONCLUSION Eight patients (21.6%) were possible PAH and 4 (10.8%) were diagnosed as SSc-PAH by RHC after the echocardiographic screening study of 37 adult SSc patients.

Aim There is growing evidence that cold-inducible RNA-binding protein (CIRP) promotes inflammatory responses. This study investigated the relationship between CIRP and rheumatoid arthritis (RA). Methods Peripheral blood and synovial fluid were collected from 15 patients with RA and from 16 patients with osteoarthritis (OA). The concentration of CIRP was measured with the sandwich enzyme-linked immunosorbent assay (ELISA). Results The concentration of serum CIRP was significantly elevated in the RA patient group (RA patients = 26.39 ± 10.48 pg/mL, OA patients = 17.14 ± 7.24 pg/mL, P = 0.009). Furthermore, the RA patient group had a significantly higher CIRP concentration than that of the OA patient group in synovial fluid (153.56 ± 108.93 pg/mL vs. 23.63 ± 16.18 pg/mL, P < 0.001). The mean synovial fluid concentration of CIRP was significantly higher than that of the serum concentration in the RA patient group (serum concentration = 26.39 ± 10.48 pg/mL, synovial fluid = 153.56 ± 108.93 pg/mL, P < 0.001). Disease Activity Score of 28 joints (DAS28)-ESR (erythrocyte sedimentation rate) and DAS28-CRP (C-reactive protein) were positively correlated with the synovial fluid concentration of CIRP (DAS28-ESR: r = 0.582, P = 0.023; DAS28-CRP: r = 0.541, P = 0.037). Conclusion The serum and synovial concentrations of CIRP in the RA patients were increased compared to the OA patients. Additionally, the synovial concentration of CIRP in RA patients correlated well with disease activity, that is, the DAS28-ESR/CRP. Based on these results, CIRP mediates inflammation and is a potential marker for synovial inflammation.

Autoimmune rheumatoid arthritis is characterized by chronic inflammation and hyperplasia in the synovial joints. Although the cause of rheumatoid arthritis is largely unknown, substantial evidence has supported the importance of immune cells and inflammatory cytokines in the initiation and progression of this disease. Herein, we demonstrated that the benzoxathiole derivative 2-cyclohexylimino-6-methyl-6,7-dihydro-5H-benzo[1,3]oxathiol-4-one (BOT-4-one) alleviated type II collagen-induced arthritis in a mouse model. The levels of pro-inflammatory cytokines are elevated in both human patients with rheumatoid arthritis and mice with collagen-induced arthritis. BOT-4-one treatment reduced the levels of pro-inflammatory cytokines in mice and endotoxin-stimulated macrophages. BOT-4-one treatment suppressed the polarization of Th1- and Th17-cell subsets by inhibiting the expression and production of their lineage-specific master transcription factors and cytokines, as well as activation of signal transducer and activator of transcription proteins. In addition, BOT-4-one inhibited mitogen-activated protein kinase and NF-kappaB signaling as well as the transcriptional activities and DNA-binding of transcription factors, including activator protein-1, cAMP response element-binding protein and NF-kappaB. Our results suggest that BOT-4-one may have therapeutic potential for the treatment of chronic inflammation associated with autoimmune rheumatoid arthritis.

B7-H3, a newly identified B7 family member, has functional duality as a co-stimulator and co-inhibitor that fine-tunes T cell-mediated immune responses. Given that B7-H3 expression on human monocytes and dendritic cells is enhanced by inflammatory cytokines, its potential inmmunoregulatory role at sites of inflammation has been suggested. Further, monocytes play crucial roles in the pathophysiology of various inflammatory disorders including autoimmune diseases; however, the immunological role of B7-H3 in rheumatoid arthritis (RA) has not been defined. Thus, we aimed to investigate the possible roles of monocyte B7-H3 in the pathogenesis of RA. Synovial monocytes, but not peripheral monocytes, in RA patients predominantly express surface B7-H3. The 4Ig isoform of B7-H3 is exclusively induced on the cell surface, whereas the 2Ig B7-H3 isoform is constitutively expressed in the intracytoplasmic region of both peripheral and synovial monocytes. B7-H3 knockdown experiments reveal that surface B7-H3 has an inhibitory effect on IFN-γ production in CD4 memory cells. Moreover, surface B7-H3 expression on synovial monocytes inversely correlates with RA clinical parameters. Our findings demonstrate that activation-induced B7-H3 expression on synovial monocytes has the potential to inhibit Th1-mediated immune responses and immunomodulatory roles affecting RA pathogenesis.

Dendritic cells (DC) are specialized antigen-presenting cells that initiate and orchestrate the body’s immune response to bridge the innate and adaptive systems. Here we report developing a standardized evaluation system for immunity-boosting functional foodstuffs using bone marrow-derived dendritic cells. A DC-based evaluation system can indicate the type, magnitude, and specificity of immune responses elicited by an immune-boosting food item. We monitored the expression profile of DC maturation markers, including CD11b, CD11c, CD40, PD-L1/L2, CD80, CD86, and MHC, by flow cytometry to define immunity indexes. These indexes are then consolidated to a 4-point scale to describe the immunogenic and tolerogenic capabilities of the immunity-boosting food candidate. Among well-known immunoregulatory materials, red ginseng extracts, fucoidan, and polysaccharide-K were found to belong to the highest grade of immunogenicity, while extract of the Ostericum root was among the most tolerogenic foods. Extracts of Peucedanum japonicum Thunberg root and Psidium guajava L. leaves also showed high tolerogenic capabilities. Our results suggest that this assay is an effective method to characterize and screen the immune boosting effects of food and pharmaceutical materials in a standardized manner.

Monocytes are important cellular effectors of innate immune defense. In humans, circulating monocytes are heterogeneous and can be divided into three distinct subsets based on CD14 and CD16 expression. The expansion of “intermediate” CD14+CD16+ monocytes is well reported in many chronic inflammatory conditions such as rheumatoid arthritis (RA). However, the mechanism underlying induction of CD16 and the role of CD16 in CD14+CD16+ monocytes remain unclear. ADP-treated autologous platelets induced CD16 expression by highly purified “conventional” CD14+CD16− monocytes. Given that platelet activation causes the release of several soluble factors, we examined an effect of cytokines, which are involved in monocyte-platelet aggregation (MPA), on CD16 induction. MPA led to significantly increased production of TGF-β and IL-6. Exogenous TGF-β and IL-6 in platelet-free culture induced CD16 expression, whereas neutralizing antibodies for TGF-β and IL-6 largely suppressed CD16 induction. Furthermore, selective inhibitors of SMAD3 and STAT3 reduced CD16 expression, indicating a critical role of TGF-β and IL-6 for CD16 induction by CD14+CD16− monocytes. Functionally, induced CD16 participates in IgG-mediated phagocytosis and the level of CD16 expression in activated platelet-treated monocytes is correlated with uptake of beads coated with FITC-labeled IgG. Lastly, CD14+CD16− monocytes treated with activated platelets differentiate into M2-like macrophage. Our findings suggest an important role of activated platelets for modulating phenotypical and functional features of monocytes in humans. This will be helpful for understanding immunological role of CD14+CD16+ in chronic inflammatory diseases.

Background Behcet’s disease(BD) is an inflammatory multi-genetic disorder with unknown etiology. In the previous study, we sequenced whole mitochondrial nucleotides from blood of 20 BD patients and 10 sex-, age-matched healthy controls, m.248A>G, m.709G>A, m.3970C>T, m.6392T>C, m.6962G>A, m.10310G>A, m.10609T>C, m.12406G>A, m.12882C>T, m.13928G>C, m.16129G>A, and m16304T>C were more frequently observed in patients group than healthy control without statistical significance. While, m.304C>A, m.3010G>A, m.4883C>T, m.5178C>A, and m.14668C>T were more frequent in control group (p=0.008, 0.026, 0.007, 0.007, and 0.026, respectively). m.16182A>C, m.16183A>C, m.16189T>C were associated with uveitis (p=0.041, 0.022, and 0.014, respectively). Objectives We performed a follow-up study to validate these possible associations in larger groups. Methods Whole blood or buffy coat were collected from 98 BD patients from four university hospitals located in Chung-Cheong district of Republic of Korea, 196 age-, sex-matched healthy controls from Konyang University Hospital. Above mentioned 20 targeted mitochondrial DNA(mtDNA) genomes were analysed using MassARRAY® system(Agena Bioscience, Inc., San Diego, CA, USA) for m.709, m.3010, m.4883, m.6392, m.6962, m.10310, m.10609, m.12406, m.12882, m.13928, and m.14668; Sanger sequencing for m.248, m.304, m.5178, m.16129, m.16182, m.16183, m.16189, and m.16304; and TaqMan-based genotyping assay(Applied Biosystems, Foster City, CA, USA) for m.3970. Results were compared with the revised Cambridge Reference Sequence (rCRS). Chi square or Fisher’s exact test were used to analyse association of mtDNA alterations between groups and between mtDNA alterations and clinical/laboratory characteristics. Results Presence of m.248A>G, m.304C>A, m.709G>A, m.3010G>A, m.3970C>T, m.4883C>T, m.5178C>A, m.6392T>C, m.6962G>A, m.10310G>A, m.10609T>C, m.12406G>A, m.12882C>T, m.13928G>C, m.14668C>T, m.16129G>A, and m16304T>C were not differentiated by the groups. However, m.16812A>C was more frequently observed in the patient group than control [22 (22.4%) vs. 24 (12.2%), p=0.061], and it was significantly associated with HLA-B51 positivity (p=0.011), arthralgia (p=0.043) and methotrexate use (p=0.02), and was not associated with uveitis in the follow-up study. Among clinical and laboratory characteristics in BD patients, thrombosis was more frequently observed in male patients than female patients [7 (22.6%) vs. 0 (0%), p Conclusions We performed a follow-up study to validate possible associations between BD and 20 mtDNA alterations. m.16812A>C could be associated with BD and its several clinical or laboratory characteristics a Korean population. References [1] Xavier JM, Shafiee NM, Ghaderi F, Rosa A, Abdollahi BS, Nadji A, et al. Association of mitochondrial polymorphism m.709G>A with Behcet’s disease. Ann Rheum Dis2011;70:1514–6. [2] Kwon MH, Joung CI. Genetic Associations of Mitochondrial DNA Polymorphisms with Behcet’s Disease in a Korean Population: A Pilot Study. J Rheum Dis 2016;23(1):23–9. Acknowledgements The current study was supported by the National Research Foundation of Korea funded by the Korean Government (grant no. NRF-2017R1C1B2008199). Disclosure of Interest None declared