Aubrey Bell

Reduced expression of CD44 on monocytes and neutrophils in systemic lupus erythematosus: relations with apoptotic neutrophils and disease activity

BACKGROUND Increased numbers of apoptotic neutrophils, and impaired monocyte/macrophage clearance of apoptotic cells, have been demonstrated in systemic lupus erythematosus (SLE). CD44 is implicated in the clearance of apoptotic neutrophils. OBJECTIVE To determine the expression of CD44 on peripheral blood monocytes and neutrophils in SLE, and examine the relations with disease activity and numbers of circulating apoptotic neutrophils. METHODS Peripheral blood was sampled from 31 patients with SLE, 19 healthy normal subjects, and 19 patients with rheumatoid arthritis (RA). Monocyte and neutrophil density of surface CD44 expression was determined by immunofluorescence labelling and flow cytometry, and results expressed as mean channel fluorescence (MCF) values. Neutrophil apoptosis was measured by morphology in 15 patients with SLE, nine with RA, and six normal subjects. RESULTS Monocyte CD44 expression was significantly lower in SLE (median MCF 4.71) than in healthy normal subjects (median MCF 5.61) and controls with RA (median MCF 5.39). Neutrophil CD44 expression was also significantly lower in SLE (median MCF 1.95) than in healthy normal subjects (median MCF 2.37) and controls with RA (median MCF 2.60). Monocyte, but not neutrophil, CD44 expression correlated negatively with the percentage of apoptotic neutrophils. There was no significant correlation of monocyte or neutrophil CD44 expression in SLE with disease activity or damage. CONCLUSIONS Monocyte and neutrophil CD44 expression is reduced in SLE, and this may contribute to the impaired recognition and clearance of apoptotic neutrophils by monocyte derived macrophages.

The CD14+ CD16+ monocyte subset in rheumatoid arthritis and systemic lupus erythematosus

Abstract Most human peripheral blood monocytes strongly express surface CD14, but not CD16 (CD14++/CD16–). A smaller group of monocytes express lower levels of CD14 and also express CD16 (CD14+/CD16+). This subgroup has different functional characteristics and is expanded in a number of disease states. We aimed to determine the percentage of circulating CD14+/CD16+ monocytes in rheumatoid arthritis and systemic lupus erythematosus (SLE) and relate this to disease measures. Peripheral blood was sampled from 31 SLE patients, 19 rheumatoid arthritis patients, and 19 healthy controls. The percentage of CD14+/CD16+ monocytes was determined by immunofluorescence labelling and dual colour flow cytometry. The percentage of CD14+/CD16+ monocytes was significantly lower in rheumatoid arthritis (median 4.90%) than in normal subjects (median 7.30%, P=0.014),and in rheumatoid arthritis than in SLE patients (median 9.40%, P=0.009). The percentage of CD14+/CD16+ monocytes in SLE was not significantly different from that in healthy subjects. This lower percentage of CD14+/CD16+ monocytes in rheumatoid arthritis may be important in the pathogenesis of this disease.

Antinucleosome antibodies in the diagnosis of systemic lupus erythematosus

Nucleosomes are fundamental units of chromatin released by internucleosomal cleavage during cell apoptosis, and nucleosomal material has been demonstrated in the surface blebs of apoptotic cells.1 Recent studies have shown the presence of antinucleosome antibodies in systemic lupus erythematosus (SLE).2,3 We measured the concentration of antinucleosome antibody present in the sera of patients with SLE and compared it with the concentration in healthy and disease control patients using a commercially available enzyme linked immunosorbent assay (ELISA) kit.nnPeripheral blood was sampled from 95 white patients with SLE (87 female, median age 47.0 years), 48 white patients with rheumatoid arthritis (RA) (41 female, median age 55.5 years), 28 white patients with fibromyalgia (23 female, median age 47.0 years), and 95 white normal healthy volunteers (64 female, median age 31.0 years). All patients with SLE fulfilled the American College of Rheumatology (ACR) diagnostic criteria.nnAn indirect solid phase immunometric assay (ELISA) was used for the quantitative determination of IgG autoantibodies to nucleosomes (Organtec Diagnostika, Mainz, Germany; antinucleosome kit), according to the …

Diclofenac inhibits monocyte superoxide production ex vivo in rheumatoid arthritis

SummaryEffects of the nonsteroidal anti-inflammatory drug, diclofenac, on stimulated monocyte superoxide production were assessed directly in vitro and following treatment of patients with rheumatoid arthritis ex vivo. Diclofenac inhibited superoxide generation provoked by serum treated zymosan (STZ) and fluoride anion (F) but not by phorbol myristate acetate (PMA) in vitro. Following patient therapy, inhibition of superoxide production occurred when STZ and PMA, but not F were used as stimuli. No changes were seen in control subjects. The contrasting profiles of inhibition seen in vitro and ex vivo suggest an indirect effect on superoxide production during clinical use of the agent. These data are consistent with the hypothesis that anti-inflammatory drugs may act in rheumatoid arthritis by inhibiting phagocyte super-oxide anion production.

Accelerated apoptosis in SLE neutrophils cultured with anti-dsDNA antibody isolated from SLE patient serum: a pilot study

Increased numbers of apoptotic neutrophils are found in SLE, related to disease activity and levels of anti-dsDNA antibody. The mechanism of increased apoptosis is not clear, but anti-dsDNA antibody has been shown to induce apoptosis in neutrophils from normal subjects and in certain cell lines. In this study, polyclonal anti-dsDNA antibody was isolated from the serum of a patient with active SLE, and was shown to substantially accelerate apoptosis in neutrophils from SLE patients as compared with neutrophils from healthy control or rheumatoid arthritis subjects.

Mercury in Hair Is Inversely Related to Disease Associated Damage in Systemic Lupus Erythematosus.

Systemic lupus erythematosus (SLE) is an autoimmune inflammatory disease, and environmental factors are proposed to exacerbate existing symptoms. One such environmental factor is mercury. The aim of this study was to investigate the relationship between exposure to mercury (Hg) and disease activity and disease associated damage in Total Hg concentrations in hair and urine were measured in 52 SLE patients. Dental amalgams were quantified. Disease activity was assessed using three indexes including the British Isles Lupus Assessment Group Index (BILAG). Disease associated damage was measured using the Systemic Lupus International Collaborating Clinics/American College of Rheumatology SLICC/ACR Damage Index. Pearson’s correlation identified a significant negative correlation between hair Hg and BILAG (r = −0.323, p = 0.029) and SLICC/ACR (r = −0.377, p = 0.038). Multiple regression analysis identified hair Hg as a significant predictor of disease associated damage as determined by SLICC/ACR (β = −0.366, 95% confidence interval (CI): −1.769, −0.155 p = 0.019). Urinary Hg was not related to disease activity or damage. Fish consumption is the primary route of MeHg exposure in humans and the inverse association of hair Hg with disease activity observed here might be explained by the anti-inflammatory effects of n-3 long chain polyunsaturated fatty acids also found in fish.

Vitamin D status and disease activity in systemic lupus erythematosus in Northern Ireland

L. C. Breslin, E. M. Duffy, H. G. Mulholland, S. A. Wright, M. S. Barnes, R. Hodd, J. M. W. Wallace and A. L. Bell Northern Ireland Centre for Food and Health, University of Ulster, Coleraine BT52 1SA, UK, Cancer Epidemiology & Prevention Research Group, Queen’s University Belfast, Royal Victoria Hospital Site, Belfast BT12 6BJ, UK and Musculoskeletal Education and Research Unit, Musgrave Park Hospital, Belfast BT9 7JB, UK

AB0408 An investigation of vitamin d status in systemic lupus erythematosus patients residing in northern ireland: its relationship with disease activity and bone mineral density

Background Vitamin D has the potential to modulate the immune system for patients with systemic lupus erythematosus (SLE)1, and which could potentially lead to improved clinical outcomes. Observational data have suggested a relationship between vitamin D and disease activity2. This observation has been confirmed by vitamin D3 supplementation studies, where positive immunological effects were reported in SLE patients3. Additionally a placebo controlled trial supplementing SLE patients with vitamin D3 identified a positive effect on inflammation and haemostatic markers as well as improvements in disease activity4. Research examining the relationship between vitamin D and bone mineral density (BMD) in SLE patients is limited, albeit a study has suggested a link between disease activity and BMD in SLE patients5 and thereby suggesting flares in disease activity has a negative effect on BMD. Objectives To examine for relationships between vitamin D and disease activity, damage and BMD. Methods A total of 52 SLE patients were recruited onto an observational study during the winter (November-March) and followed up during the summer months (June-July) (n=50). Total 25-hydroxyvitamin D (25(OH)D) concentration was measured using the liquid chromatography mass spectrometry method (MassChrom®, Chromsystems Gmbh, Heimburgstrasse, Germany) and BMD was measured at the lumbar spine and femur by dual energy X-ray absorptiometry (Lunar iDXA™, UK). Disease activity and damage were assessed using Systemic Lupus Activity Measure (SLAM), British Isles Lupus Assessment Group (BILAG), Systemic Lupus Erythematosus Disease Activity Index (SELENA SLEDAI) and Systemic Lupus International Collaborative Clinics/American College of Rheumatology (SLICC/ACR). Results Mean (SD) 25(OH)D concentrations in winter (26.9 (16.2) nmol/L) and summer (28.7 (16.7) nmol/L) were not different (P=0.439), nor were differences observed between disease activity and damage. During the winter, but not during the summer, vitamin D was a significant predictor of BILAG (r=-0.307; P=0.027) and SELENA SLEDAI (β=-0.074; SE=0.036; P=0.044), controlling for age, BMI and medications. Osteoporosis and osteopenia was present in some 4 (8%) and 21 (45%) SLE patients respectively and there were no relationships observed between disease activity and BMD. Conclusions Vitamin D inadequacy was prevalent throughout the year and undiagnosed osteoporosis was apparent in this cohort, and, therefore, suggesting routine screening of both is warranted. Wintertime vitamin D status was a predictor of disease activity at that time of year. There was no relationship, however, observed between disease activity and BMD. The lack of seasonal variability in vitamin D status might suggest sun avoidance by the SLE patients; emphasising the importance of obtaining adequate vitamin D intake from the diet and supplements for SLE patients. References Marques et al. Revista Brasileira de Reumatologia. 2010;50(1):67-80. Breslin, et al. Proceedings of the Nutrition Society. 2011 Nov;70(4):399-407. Terrier, et al. Arthritis Research and Therapy. 2012 Oct 17;14(5). Abou-Raya A, et al. Journal of Rheumatology, 2012. Zhang et al. Zhonghua Yi Xue Za Zhi, 2012 Sep 4;92(33):2331-4. Disclosure of Interest None Declared

THU0314 Plasma IL-6 Levels Correlate with Ultrasound Measures of Disease Activity in Lupus Arthritis

Background The role of specific cytokines in lupus arthritis has not been elucidated1. This is in stark contrast to rheumatoid arthritis (RA) where an abundance of research has culminated in an advancing era of novel biologic drugs. Objectives To analyse the cytokine profile in SLE patients with erosive, non-erosive arthritis and arthralgia as classified by ultrasound (US). Methods 50 SLE patients, and 40 RA patients had an US scan of their hand as per standardised protocols2-3. US scores per patient were expressed per joint and as a total ‘ultrasound activity’ score, (sum of Power Doppler (PD) and Grey Scale Synovial Hypertrophy scores in all joints) and a total erosion score. SLE disease activity was assessed (BILAG and SELENA SLEDAI). Plasma levels of IL-6, TNF-alpha and BLyS were measured using sandwich ELISA kits (Quantikine® kits, R & D). Results On the basis of the ultrasound results the SLE patients were divided into three groups, those with erosive arthritis (n = 24), those with non-erosive arthritis (n = 14) and those with a normal ultrasound scan (n = 12). CRP and IL-6 levels between the erosive lupus group and the RA group were not significantly different (p = 0.3 and p = 0.2 respectively). Conclusions This is the first study to examine levels of specific cytokines in a cohort of SLE patients stratified in terms of joint disease by ultrasound where the most significant finding is that IL-6 levels correlated with both clinical and ultrasound measures of arthritis disease activity. There is preliminary evidence that IL-6 blockade may be a potential treatment for SLE4 and if arthritis specific outcome measures were included in future clinical trials more robust evidence could be generated. References Jacob N & Stohl W. Cytokine disturbances in Systemic Lupus Erythematosus. Arthritis Research & Therapy 2011; 13 (4): 244 Wakefield RJ et al. Musculoskeletal ultrasound including definitions for ultrasonographic pathology. J Rheum 2005;32(12):2485-87. Szkudlarek M et al. Interobserver agreement in ultrasonography of the finger and toe joints in rheumatoid arthritis. A & R 2003;48(4):955-62. Illei G et al. Tocilizumab in systemic lupus erythematosus: data on safety...... A & R 2010;62(2):542-52. Disclosure of Interest None Declared

THU0315 Rhupus’ Arthritis - An MRI and Ultrasound Perspective

Background The limited application of Ultrasound(US) and MRI to date in SLE is revealing a higher percentage of erosive disease than previous estimates1. Such erosive arthropathy in lupus when associated with rheumatoid factor (RF) or anti-CCP antibody (ACPA) is often referred to as ‘rhupus’ to indicate a mixture of the character of rheumatoid disease (RA)3. Objectives To evaluate SLE hand symptoms from both an Ultrasound and an MRI perspective. Methods 50 SLE patients with joint symptoms and 40 RA patients had a detailed US scan (Grey-scale (GSSH) and Power Doppler (PD)) of their hand as per standardised protocols3-4. 32 of the SLE patients also had a contrast enhanced MRI of their hand. Results Imaging findings related to inflammatory arthritis were detected in 38 (76%) lupus patients. 18 (36.2%) SLE patients had erosive disease at either the wrist or MCP joints on US, an additional 4 patients had erosions on MRI at the wrist which were not visualised on US. One patient had an erosion at the 2nd MCP joint on US which was not seen on MRI. On the basis of the combined US/MRI results the SLE patients were divided into erosive, non-erosive arthritis and arthralgia. There was no difference in median (IQR) CRP between the RA group (5.6 (2.1, 22) mg/dl and the erosive lupus group (4.2 (1.5,7.0)), but there was a significant difference between the RA group and the non-erosive lupus group (1.6 (1.1,4.7))(p = 0.004). CRP also correlated with total erosion score in the lupus group (p = 0.03). There was a higher mean total ultrasound activity score (sum of GSSH and PD) in the erosive lupus group as compared to the non-erosive group (p = 0.03). Five erosive SLE patients had a positive ACPA or RF. A higher percentage of patients in the erosive lupus group (41.6%) had positive anti-Ro antibodies as compared to the non-erosive lupus (28.5%) groups (P <0.001). Conclusions This is the first study to combine US and MRI in the assessment of SLE patients and to provide information on a cohort of symptomatic lupus arthritis patients where the most significant finding is that 44% had erosive disease, only half ( 22.3%) of which were ACPA or RF positive. Within this sub-group of erosive lupus arthritis patients the most obvious discriminating features were a higher ultrasound activity score, a higher CRP and a higher prevalence of anti-Ro antibodies. This association with CRP and lupus arthritis has been previously described albeit in very small studies5. Advanced imaging techniques may have an impact on the perception of SLE arthritis that will ultimately influence treatment. The ability to assign SLE patients into categories of which the natural history of joint disease progression were known and to target those patients who warrant aggressive treatment would be invaluable in terms of therapeutic rationale. References Wright S et al. Hand arthritis in SLE: an ultrasound pictorial essay. Lupus 2006;15(8) Fernandez A, et al. Lupus arthropathy: a case series opatients with rhupus. Clinical Rheum 2006 25(2) Szkudlarek M et al Interobserver agreement in ultrasonography of the finger and toe joints in rheumatoid arthritis. A & R 2003;48(4) Wakefield RJ et al. Musculoskeletal ultrasound including definitions for ultrasonographic pathology. J Rheum 2005;32(12) Spronk PE et al. Patients with SLE and Jaccoud’s arthropathy: a clinical subset with an increased CRP response? ARD 1992;51(3) Disclosure of Interest None Declared