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Autoimmune connective tissue diseases arise in a stepwise fashion from asymptomatic preclinical autoimmunity. Type I interferons have a crucial role in the progression to established autoimmune diseases. The cellular source and regulation in disease initiation of these cytokines is not clear, but plasmacytoid dendritic cells have been thought to contribute to excessive type I interferon production. Here, we show that in preclinical autoimmunity and established systemic lupus erythematosus, plasmacytoid dendritic cells are not effector cells, have lost capacity for Toll-like-receptor-mediated cytokine production and do not induce T cell activation, independent of disease activity and the blood interferon signature. In addition, plasmacytoid dendritic cells have a transcriptional signature indicative of cellular stress and senescence accompanied by increased telomere erosion. In preclinical autoimmunity, we show a marked enrichment of an interferon signature in the skin without infiltrating immune cells, but with interferon-κ production by keratinocytes. In conclusion, non-hematopoietic cellular sources, rather than plasmacytoid dendritic cells, are responsible for interferon production prior to clinical autoimmunity., Type I interferon drives autoimmune pathology in SLE and has been assumed to come predominantly from plasmacytoid dendritic cells (pDCs). Here, the authors show that prior to the onset of SLE, pDCs lose multiple immunogenic functions and, instead, non-hematopoietic cells such as keratinocytes are a major source of type I interferons.

Objective: Type I interferon (IFN ) responses are broadly associated with autoimmune diseases, including systemic lupus erythematosus (SLE ). Given the cardinal role of autoantibodies in SLE , this study was undertaken to investigate whether the findings of a B cell–specific IFN assay correlate with SLE activity. Methods: B cells and peripheral blood mononuclear cells (PBMC s) were stimulated with type I IFN and type II IFN . Gene expression was analyzed, and the expression of pathway‐related membrane proteins was determined. A flow cytometry assay for tetherin (CD 317), an IFN ‐induced protein ubiquitously expressed on leukocytes, was validated in vitro and then clinically against SLE diagnosis, plasmablast expansion, and the British Isles Lupus Assessment Group (BILAG ) 2004 score in a discovery cohort (n = 156 SLE patients, 30 rheumatoid arthritis [RA ] patients, and 25 healthy controls). A second, longitudinal validation cohort of 80 SLE patients was also evaluated for flare prediction. Results: In vitro, a close cell‐specific and dose‐response relationship between type I IFN –responsive genes and cell surface tetherin was observed in all immune cell subsets. Tetherin expression on multiple cell subsets was selectively responsive to stimulation with type I IFN compared to types II and III IFN s. In patient samples from the discovery cohort, memory B cell tetherin showed the strongest associations with diagnosis (SLE :healthy control effect size 0.11 [P = 0.003]; SLE :RA effect size 0.17 [P < 0.001]), plasmablast numbers in rituximab‐treated patients (R = 0.38, P = 0.047), and BILAG 2004. These associations were equivalent to or stronger than those for IFN score or monocyte tetherin. Memory B cell tetherin was found to be predictive of future clinical flares in the validation cohort (hazard ratio 2.29 [95% confidence interval 1.01–4.64]; P = 0.022). Conclusion: Our findings indicate that memory B cell surface tetherin, a B cell–specific IFN assay, is associated with SLE diagnosis and disease activity, and predicts flares better than tetherin on other cell subsets or whole blood assays, as determined in an independent validation cohort.

ObjectiveTo evaluate clinical, interferon and imaging predictors of progression from ‘At Risk’ to autoimmune connective tissue diseases (AI-CTDs).MethodsA prospective observational study was conducted in At-Risk of AI-CTD (defined as antinuclear antibody (ANA) positive; ≤1 clinical systemic lupus erythematosus (SLE) criterion; symptom duration Results118 individuals with 12-month follow-up were included. Of these, 19/118 (16%) progressed to AI-CTD (SLE=14, primary Sjogren’s=5). At baseline, both IFN scores differed among At-Risk, HCs and SLE groups (pConclusionA two-factor interferon score and family history predict progression from ANA positivity to AI-CTD. These interferon scores may allow stratification of individuals At-Risk of AI-CTD permitting early intervention for disease prevention and avoid irreversible organ damage.

Background Cognitive dysfunction is a common problem in autoimmune connective tissue diseases such as SLE that can be screened for using the Montreal Cognitive Assessment (MoCA). The causes of cognitive dysfunction are poorly understood. They may include immune-mediated neurological dysfunction (which should be targeted using immunosuppression) or traditional cardiovascular risk factors (which may be treated as in non-SLE patients). The purpose of this study was to explore these counter-hypotheses in patients with established autoimmune disease by analysing conventional measures of autoimmunity, cardiovascular risk, as well as validated scores for interferon status. Since cumulative organ damage and toxicity of therapy may affect these patients, we also included a cohort of At Risk individuals as previously described.1 Methods We assessed three cohorts: (1) patients with established AI-CTD (SLE, Sjogrens syndrome or undifferentiated CTD >12 months); (2) at risk individuals referred to secondary care due to ANA +and symptoms suggestive of AI-CTD less than 12 months duration; (3) age and sex matched healthy controls. Cognitive dysfunction was tested using the MoCA. Cardiovascular risk was assessed by recording diabetes, hypertension, previous angina or AMI, atrial fibrillation and cholesterol. Type I interferon activity was assessed using a validated two-score system for IFN status previously described.2 Results As expected, the number of patients with an abnormal MOCA score was greater in AI-CTD and At-Risk individuals than in the healthy controls (HC: 20%; At Risk: 39%; SLE: 34%). Also as expected, the IFN scores varied significantly between these groups (p=0.046, F=4.66). We compared parameters between individuals with normal and abnormal MoCA scores within each group and in all groups. Results are shown in table 1. In patients with an AI-CTD, the cognitive function assessed wasnt associated to any of the immune-related but associated with the presence of a cardiovascular risk factor (p=0.04) while CD was associated with anxiety and depression in at risk individuals (p=0.047). A relationship between CD and level of education, gender and current work was also observed. Conclusions In this exploratory study we identified an association between conventional cardiovascular risk factors and cognitive dysfunction. However there was no association between any of the immune parameters and MoCA score. Prevention of cognitive dysfunction in SLE should focus on early identification and treatment of cardiovascular risk. Funding Source(s): None References Md Yusof, et al. ARD 2018. El-Sherbiny, et al. Sci Rep 2018.

ObjectiveType I interferon (IFN-I) responses are broadly associated with autoimmune disease including SLE. Given the cardinal role of autoantibodies in SLE, we investigated whether a B lineage cell-specific IFN assay might correlate with SLE activity.MethodsB cells and PBMCs were stimulated with IFN-I and IFN-II. Gene expression was scrutinised for pathway-related membrane protein expression. A flow-cytometric assay for tetherin (CD317), an IFN-induced protein ubiquitously expressed on leucocytes, was validated in vitro then clinically against SLE diagnosis, plasmablast expansion, and BILAG-2004 score in a discovery cohort (156 SLE; 30 RA; 22 healthy controls). A second longitudinal validation cohort of 80 patients was also evaluated for SLE flare prediction.ResultsIn vitro, a close cell-specific and dose-responsive relationship between IFN-I responsive genes and cell surface tetherin in all immune subsets existed. Tetherin expression was selectively responsive to the IFN-I compared to IFN-II and -III. In the discovery cohort memory B-cell tetherin was best associated with diagnosis (SLE/HC: effect size=0.11, p=0.003;SLE/RA: effect size=0.17, pConclusionMemory B cell surface tetherin, a reflection of cell-specific IFN response in a convenient flow cytometric assay, was associated with SLE diagnosis, disease activity and predicted flares better than other cell subsets or whole blood assays in independent validation cohorts.

Systemic Sclerosis (SSc) is an autoimmune disorder characterized by microvascular injury and diffuse fibrosis of the skin and internal organs. While macrovascular disease and higher risk for cardiovascular events are well documented in other systemic rheumatic diseases such as rheumatoid arthritis and systemic lupus erythematosus, the presence and extent of atherosclerosis among patients with SSc is yet to be established. Primary cardiac involvement, due to impairment of coronary microvascular circulation and myocardial fibrosis, considerably affects prognosis and life expectancy of individuals with SSc, representing one of the leading causes of death in this population. On the other hand the existence and prevalence of atherosclerotic coronary disease remains an issue of debate as studies comparing structural and morphological markers of atherosclerosis and cardiovascular events between SSc patients and the general population have yielded controversial results. The aim of this review is to summarize recent literature about the prevalence of cardiovascular disease in SSc, review the surrogate markers of CVD that have been evaluated and examine whether common pathogenic mechanisms exist between SSc and macrovascular disease.

Background:We previously reported results from the first 118 “At-Risk” of autoimmune connective tissue disease (AI-CTD) individuals (i.e. ANA positivity, non-specific symptoms of ≤1 year and treatment naïve). At 1 year, 16% progressed to meet classification criteria for an AI-CTD. This was predicted by high baseline interferon (IFN) Score B and family history of RMD[1].However, some may have progressed at later time points, or had clinically significant disease despite not meeting diagnostic criteria. Longer term outcomes, baseline and follow up flow cytometry biomarkers were never reported.Objectives:(i)Describe detailed analysis of 3-year follow-up data of the At-Risk cohort(ii)Evaluate flow cytometric biomarkers as predictors of these outcomes(iii)Analyse follow up biomarkersMethods:We conducted a prospective observational longitudinal study of At-Risk individuals in Leeds (n=150). Patients were assessed at baseline, then annually for 3 years. Depending on diagnostic criteria and need for therapy, patients were grouped as follows:Absolute non-progressors (no clinical diagnostic criteria)Undifferentiated CTD (U-CTD) (≥1 clinical criteria at baseline persisting at follow-up but not meeting criteria). This group was subdivided into those who required treatment with an immunosuppressant (IS) excluding antimalarials and those who did notYear 1 progressors (meeting criteria for an RMD by 1 year)Late progressors (meeting criteria for AI-CTD beyond 1 year follow-up).Bloods were analysed at baseline and 1 year for two IFN-stimulated gene expression scores previously described[2], monocytes and subsets of B and T cells using flow cytometry. Association between clinical criteria, biomarkers at baseline and long term outcomes were tested using ANOVA.Results:3 year follow up data was available in 147/150 patients. Outcomes were: Absolute non-progressors: 63/147 (43%); U-CTD: 54/147 (37%); Year 1 progressors: 21/147 (14%) [SLE=18; pSS=3]; Late progressors (in years 1-2): 9/147 (6%) [SLE=7; pSS=2]. None progressed or required IS initiation beyond the first 2 years of follow-up. In U-CTD group, 7/54 (13%) were prescribed an IS.This work describes a larger group of 36/147 (24%) At-Risk individuals who developed clinically significant disease (CSD: progressors or need for IS) versus clinically non-significant disease (CNSD: absolute non-progressors or UCTD not needing IS).Analysis of baseline biomarkers between CSD and CNSD confirmed a significant difference in IFN Score B (mean difference -0.74, p = 0.027), but not IFN Score A (mean difference -0.68, p = 0.15). In flow cytometry analysis, there was also a significant difference in percentage monocytes (mean difference -4.09, p = 0.004) but no other subset. Absence of clinical criteria at baseline did not predict clinical outcome, and no one clinical criterion had greater predictive value.In follow up samples we noted a significant reduction in expression of IFN Score B in both groups, regardless of whether they received antimalarials or IS therapy.Conclusion:Here we report findings of a larger group of 24% At-Risk individuals who developed CSD (progressors and patients who did not meet criteria but needed IS therapy). These results provide a more complex picture of IFN activity in the initiation of SLE than previously suspected. First, we confirm that a specific subset of ISGs rather than a classic IFN signature predicts progression. Second, the reduction in IFN-Score-B in both groups suggests that IFN Score B activity is a transient phenomenon, playing a greater role in disease initiation than in disease maintenance.References:[1]Md Yusof MY, Psarras A, El-Sherbiny YM, et al. Prediction of autoimmune connective tissue disease in an at-risk cohort: prognostic value of a novel two-score system for interferon status. Ann Rheum Dis. 2018 Oct;77(10):1432-1439.[2]El-Sherbiny YM, Psarras A, Md Yusof MY, et al. A novel two score system for interferon status segregates autoimmune diseases and correlateswith clinical features. Sci Rep. 2018 Apr 11;8(1):5793.Disclosure of Interests:Sabih-Ul Hassan: None declared, Zoe Wigston: None declared, Antonios Psarras: None declared, Katie Dutton: None declared, Md Yuzaiful Md Yusof: None declared, Edward Vital Grant/research support from: AstraZeneca, Roche/Genentech, and Sandoz, Consultant of: AstraZeneca, GSK, Roche/Genentech, and Sandoz, Speakers bureau: Becton Dickinson and GSK

Measurement of type I interferon (IFN-I) has potential to diagnose and stratify autoimmune diseases, but existing results have been inconsistent. Interferon-stimulated-gene (ISG) based methods may be affected by the modularity of the ISG transcriptome, cell-specific expression, response to IFN-subtypes and bimodality of expression. We developed and clinically validated a 2-score system (IFN-Score-A and -B) using Factor Analysis of 31 ISGs measured by TaqMan selected from 3-IFN-annotated modules. We evaluated these scores using in-vitro IFN stimulation as well as in sorted cells then clinically validated in a cohort of 328 autoimmune disease patients and healthy controls. ISGs varied in response to IFN-subtypes and both scores varied between cell subsets. IFN-Score-A differentiated Systemic Lupus Erythematosus (SLE) from both Rheumatoid Arthritis (RA) and Healthy Controls (HC) (both p

Systemic sclerosis (SSc) is a connective tissue disease which is characterized by endothelium dysfunction, inflammation and fibrosis. Although scleroderma is often presented as an overlap syndrome with other autoimmune rheumatic diseases, the development of large vessel vasculitis in patients with SSc is considered extremely rare, since only three case reports have thus far been reported in English literature. Herein, we report a 65-year-old woman with a long-standing history of systemic sclerosis who developed giant cell arteritis, eight years after initial diagnosis.

Background and objectives Type I interferons (IFN-I) are crucial to systemic lupus erythematosus (SLE) pathogenesis. However, IFN-I activity, as measured by interferon-stimulated gene (ISG) expression, has an unclear relationship with clinical features of disease. Incomplete lupus erythematosus (ILE) describes individuals who have new onset of features suggestive of SLE, but do not fulfil diagnostic criteria. Up to 20% of these patients eventually progress to SLE. The objective of this study is to compare ISG expression between patients with ILE and SLE to understand the role of IFN in onset of clinical features of SLE. Materials and methods Expression of 33 ISGs was measured using qPCR in PBMCs from individuals with SLE (n = 54), ILE (n = 27), and healthy controls (HC; n = 14). SLE was defined using 2012 ACR/SLICC criteria. ILE was defined as ANA +ve, 1–2 clinical ACR/SLICC criteria, and symptom duration Results FA on SLE patients indicated two factors explaining 80% of the data variance. In total, 16 and 14 genes loaded onto Factors F1 and F2 respectively. The majority of variability was explained by F1; however, F2 appeared more relevant to the presence of fully established SLE. F1 and F2 were significantly different between patient groups (p = 0.005 and p = 0.044 respectively). F1 was similarly high in both SLE [SLE:HC=4.22 (1.80, 9.88), p = 0.001] and ILE [ILE:HC=2.96 (1.20, 7.32), p = 0.019]. In contrast, F2 was increased only in SLE [SLE:HC=1.38 (0.95, 2.00), p = 0.086] but not in ILE [ILE:HC=1.02 (0.69, 1.51), p = 0.917]; a significant difference was observed between SLE and ILE patients [SLE:ILE=1.35 (1.04, 1.77), p = 0.026]. Conclusions IFN-I activity is present in ILE. However, the majority of measured ISG expression (F1) cannot distinguish ILE and SLE. F1 represents genes that distinguish healthy individuals, but show little variation at different stages of the disease development. We define a subset of ISGs (F2) whose expression is only increased in patients with confirmed clinical SLE. ISG expression is not unidimensional: qualitative differences in expression of distinct ISGs contribute to clinical progression after disease initiation.