mice treated with monoclonal anti-TNF: 5/15 (33%) with TN3 (p=0.03/ controls), 4/12 (33%) with ADA (p=0.054/ controls), 0/15 with ETA and 1/22 (5%) in controls. Conclusions Higher mortality and increased risk of lymphoma were observed in BAFF Tg mice treated with monoclonal anti-TNF compared to etanercept. This result may be linked either to the different mechanism of action between the soluble receptor and the monoclonals or to a difference of trough level observed in the different groups even if higher levels of ADA was mandatory given the difference of effect on mouse TNF. This study demonstrates the negative impact of a prolonged anti-TNF treatment on the risk of lymphoma in the context of BAFF increase. Disclosure of interest None declared P029ABSTRACT WITHDRAWN

Background:The insulin-like growth factor 1 receptor (IGF1R) signalling mediates numerous developmental processes acting through downstream adaptor molecules IRS1/2, which activate Akt and inhibit the family of forkhead box class O (FoxO). Inhibition of IGF1R signalling alleviates rheumatoid arthritis (RA) (Erlandsson et al., 2017), however, the role of IGF1R signalling in the regulation of immune function is poorly understood.Objectives:To investigate the link between IGF1R signalling and antigen presentation in experimental arthritis.Methods:Arthritis was induced by immunising Balb/c mice with methylated bovine serum albumin (mBSA, n=18) and DBA/1 mice with type II collagen (CII, n=18). The mice were treated with a synthetic IGF1R inhibitor NT157 or with short hairpin RNA targeting IGF1R (shIGF1R) from the day prior to immunisation. Controls were treated with cyclodextrine vehicle/ non-targeting (nt)RNA, respectively. Flow cytometry was used for spleen cell phenotype. Antibody levels were measured by ELISA. Immunohistochemistry (IHC) of spleen was performed for assessment of marginal zone (MZ) and location of pS612IRS1+ and pS256FoxO1+ cells. IHC images were acquired by fluorescent confocal microscopy, and analysed using ZEN2009 and Cell Profiler soft ware.Results:The inhibition of IGF1R resulted in an 80% increase in MZ area in NT157-treated mice compared to controls (p=0.0001). This was supported by a significant increase of CD21+ (p=0.034) and CD23+ cell populations (p=0.00059), both among the CD19+ B cells and antigen-presenting MHCII+CD19- cells, implying that IGF1R expression regulates the populations of MZ and follicular cells. Additionally, there was a strong positive correlation between the decrease of IGF1R+ and ICOSL+ population on CD21+ cells (r=0.70, p=0.0071), which retained them in the MZ and prolonged communication with macrophages. Insufficient feedback from ICOSL- B cells limited expression of CXCR5 on CD4 cells. The IHC analysis displayed that, IGF1R inhibition led to abundance of inactivate pS612IRS1+ and pS256FoxO1+ cells within the MZ, compared with controls (p=0.0002). Alongside the increase of IgM+ B cell population (p=0.0022), we observed significant increase in number of antigen-presenting F4/80+ cells (p=0.043) and MARCO expression (p=0.043) after IGF1R intervention. Finally, the NT157- treated mice displayed a significant pleiotropic increase in IgM autoantibody production, with anti-CCP IgM (p=0.027), RF-IgM (p=0.0085), anti-DNA IgM (p=0.066) and in total IgM (p=0.027) levels, which correlated positively with pS256FoxO1+ cells (r=0.51, p=0.03). Levels of IgG were not changed.Conclusion:We show that IGF1R signalling is important for immune cell communication after antigen challenge. IGF1R controls ICOSL dependent trafficking of B cells through the MZ and facilitates interaction with T cells. Retention of B cells in the MZ tips the balance from T cell to macrophage-dependent processes, which permits the formation of autoantibody producing B cells.References:[1]Erlandsson, M., et.al., 2017. IGF-1R signalling contributes to IL-6 production and T cell dependent inflammation in rheumatoid arthritis. Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease, 1863(9), pp.2158-2170.Disclosure of Interests:None declared

IntroductionSmoking is a risk factor for the development of rheumatoid arthritis (RA) and associates with positivity for disease specific anti cyclic citrullinated peptide (aCCP) antibodies. It is not known exactly how smoking promotes autoimmunity but we have previously demonstrated that smoking limits the expression of the T cell co-inhibitory receptor programmed death-1 (PD-1).1 ObjectivesTo investigate if smoking can interfere with the interaction between PD-1 and its ligand by influencing the serum levels of soluble PD-1 ligand 1 (sPD-L1).MethodsSerum samples were collected from 254 RA patients and 168 healthy controls with known smoking status and analysed with ELISA for levels of sPD-L1 and inflammatory cytokines IL-6 and IL-1b. Fc-receptor mRNA expression analysis of peripheral blood monocytes (PBMC) from a group of 10 healthy controls and 15 RA patients was done by qPCR.ResultsIn RA patients current smokers had 5 times lower median serum levels of sPD-L1 compared to never smokers (p=0.027). This difference persisted in former smokers that quit smoking <25 years ago (p=0.0086). Expectedly, sPD-L1 were associated with high serum levels of inflammatory markers IL-6 (over 5.5 pg/ml, p=0.0006) and IL-1b (over 4 pg/ml, p=0.0005), however the level of these markers was not affected by the smoking status of the patients. We found 14 times lower sPD-L1 levels in smoking RA patients that did not receive TNF-inhibitors (p=0.0092), but treatment with TNF-inhibitors normalised levels of sPD-L1.Furthermore, aCCP positivity in RA patients was associated with higher levels of sPD-L1 (p=0.0036). We speculate that the presence of antibodies might influence the levels of sPD-L1 through the stimulation of Fc-receptors expressed by PD-L1 producing cells. In PBMC depleted of T cells, we saw that smokers had lower mRNA expression of the stimulatory FcgRIIIA (p=0.028) and predominance of the inhibitory FcgRIIB in the FcgRIIB/FcgRIIIA ratio (p=0.0004).ConclusionsSmoking decreases the serum levels of the inflammation limiting protein sPD-L1, but levels were restored by treatment with TNF-inhibitors. aCCP positive RA patients had higher levels of sPD-L1, possibly due to activation of Fc-receptors expressed by PD-L1 producing cells.Reference. Wasén C, et al. J Autoimmun2017.Disclosure of interestNone declared

IntroductionIn addition to inflammation of the joints, rheumatoid arthritis (RA) has a neurological part consisting of pain, fatigue, depression and cognitive deterioration. These symptoms are critical for the patients’ ability to cope with daily life, but are not alleviated completely with modern antirheumatic drugs. Sufficient IGF1R signalling is important for neurogenesis in the hippocampus. Its misbalance correlates with depression and anxiety.ObjectivesThe aim of this study was to evaluate the pathological changes in the brain during experimental RA and investigate their connexion to IGF1R.MethodsCharacteristics of pain, depression and anxiety were collected among 214 RA patients and analysed in relation to IGF1R expression in WBC. Experimental RA was induced by immunisation with collagen II. Inhibition of IGF1R was achieved by injection of shRNA-producing lentiviral construct. The behavioural pattern of each mouse was recorded by filming. The hippocampus was analysed morphometrically, and gene and protein expression were analysed by qPCR and immunohistochemistry, respectively.ResultsThe RA patients’ perception of depression and anxiety was associated with high IGF1R expression in WBC. This group of patients was also less physically active. In experimental RA, an enrichment of IBA1 +microglia and high expression of CD68 and IL-1b was found in the hippocampus. This was followed by an increased density of IGF1R+cells in cornu ammoni, and a decreased neurogenesis by limited expression DCX in the subgranular layer of the dentate gyrus. This results in a significant reduction of the hippocampus area. These changes in the brain were associated with immobility in RA mice. Treatment with shRNA targeting IGF1R improved arthritis, but led to increased immobility.ConclusionsRA induces remote inflammation in the hippocampus reducing neurogenesis and physical activity. The neurological symptoms in patients and in experimental RA are connected to IGF1R expression and signalling, and further expands our knowledge of neurological processes in RA.Disclosure of interestNone declared

Background Presence of antibodies against citrullinated peptides (anti-CCP2) and increased levels of cytokines and chemokines precedes the development of rheumatoid arthritis (RA) by several years [1, 2]. Recently, the proteins survivin and Fms-like tyrosine kinase 3 ligand (Flt3L) have been associated with RA and RA associated joint damage [3, 4]. Objectives To investigate the potential of survivin and Flt3L as predictors of RA and their relationships to cytokines and anti-CCP antibodies in blood samples from individuals before onset of symptoms of RA. Methods This study includes 47 individuals sampled before onset of symptoms of RA (median 2.5 years (IQR 1.1-5.6) and 155 population controls matched for sex and age, all donors to the Medical Biobank of Northern Sweden. 36 of the pre-symptomatic individuals were also sampled at the time of RA diagnose (ACR criteria 1987). Levels of survivin and Flt3L were measured using sandwich ELISAs (both, R&D Systems, Minneapolis, MN). Anti-CCP antibodies was analyzed using ELISA (Euro-Diagnostica AB, Malmö, Sweden) and 29 cytokines, and chemokines (IL-1β, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12, IL-13, IL-15, IL-17, eotaxin, IL-1Ra, bFGF, G-CSF, GM-CSF, IFNγ, IP-10, MCP-1, MIP-1α, MIP-1β, PDGFBB, TNFα, VEGF, Mig, MIF and IL-2Rα by multiplex detection (Bio-Rad, Hercules, CA). The cut-off levels were for survivin 450 pg/mL, and for Flt3L 130 pg/mL. Results The levels of survivin were increased in the pre-symptomatic individuals compared with the controls (p=0.003) whilst the levels of Flt3-ligand (p=0.21) were similar. The frequency of survivin in the pre-symptomatic individuals was increased compared with controls (36.2 vs. 14.2% p=0.001). The odds ratio (OR) for predicting disease development in individuals with survivin levels above cut-off was 3.4 (95%CI 1.6-7.2). The frequencies of survivin and Flt3L were increased in RA patients compared with controls (both, p<0.0001. OR12.1 [95%CI, 5.3-27.6] and OR11.0 [3.9-30.9], respectively). Anti-CCP positive pre-diseased individuals and RA-patients had significantly higher concentrations of survivin compared with those being negative. After correction for the number of comparisons, IL-1β, GM-CSF in pre-symptomatic individuals was correlated with survivin and IL-1α, IL-10, eotaxin and TNF-α was correlated with Flt3L, while IL-2, IL-9 and IL12 was correlated with both survivin and Flt3L in the pre-symptomatic individuals. Conclusions The proto-oncogene survivin was increased in individuals before onset of symptoms of RA and it was related to cytokines suggesting its role in the events preceding development of the disease. References Rantapää-Dahlqvist S et al. Antibodies against cyclic citrullinated peptide and IgA rheumatoid factor predict the development of rheumatoid arthritis. Arthritis Rheum. 2003; 48:2741-9. Kokkonen H, et al. Up-regulation of cytokines and chemokines predates the onset of rheumatoid arthritis. Arthritis Rheum. 2010; 62:383-91. Svensson B et al. Increased expression of proto-oncogene...