M. Vázquez-Del Mercado

TH1/TH2 cytokine profile, metalloprotease-9 activity and hormonal status in pregnant rheumatoid arthritis and systemic lupus erythematosus patients

During the course of rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE), several immune and neuroendocrine changes associated with pregnancy may exert positive (amelioration) or negative (exacerbation) effects on the clinical outcome. In order to shed light on the mechanisms underlying these responses, we performed a prospective longitudinal study in RA and SLE pregnant women, including healthy pregnant women as a control group. Cytokine messenger RNA (mRNA) expression assessed by quantitative competitive polymerase chain reaction (PCR) in peripheral blood mononuclear cells (PBMC), cytokine levels and lymphocyte proliferation responses (LPR) following phytohaemagglutinin (PHA) stimulation of PBMC, plasma metalloprotease‐9 activity (MMP‐9) and hormonal status during pregnancy were determined. TNFa was the most abundant cytokine mRNA expressed in PBMC in all groups studied (healthy pregnant women, RA and SLE pregnant patients). However, a general TH2 response reflected by high IL‐10 levels was found in RA, as well as SLE, patients. A significant change in IFN‐γ was observed in RA patients but only during the first trimester of pregnancy. This compared with a major TH1 response in healthy pregnant women. Interestingly, our study showed a homogeneous hormonal pattern in RA and SLE patients. Although decreased cortisol levels were observed in all patients studied, this is possibly related to the remission of disease activity status brought about by steroid treatment before and during pregnancy. In summary, we suggest that complex immune and hormonal networks are involved in pregnancy and that rheumatic diseases are very dynamic immune processes that cannot be described with a clear‐cut cytokine profile. Furthermore, the observations in this study may reflect treatment‐related immune effects more than those associated with disease.

Polymorphism of the β3-adrenergic receptor and lipid profile in patients with rheumatoid arthritis and systemic lupus erythematosus treated with chloroquine

We investigated the effect of beta 3-adrenergic receptor (β3AR) polymorphism on lipid profiles in patients with rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) treated with chloroquine. One hundred sixty-eight subjects were classified into three groups: 61 RA patients, 57 SLE patients, and 50 healthy subjects. All patients fulfilled the 1987 and 1982 classification criteria for RA and SLE, respectively, of the American College of Rheumatology. Demographic data and clinical characteristics of the patients were registered. Fasting lipid profile determination and leukocyte genomic DNA isolation from peripheral blood was performed in all the participants. Screening of the β3-AR gene polymorphic region (exon 1) was done by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. Quantitative and qualitative variables were analyzed using analysis of variance (ANOVA) with the LSD and χ2 tests, respectively. An association between the arg64/arg64 β3-AR genotype and high levels of triglycerides (TG) and very low-density lipoprotein cholesterol (VLDL-c) was found in three RA patients (P=0.01), two of them taking chloroquine. Arg64/arg64 β3-AR polymorphism may contribute to increased TG and VLDL-c in RA patients, independently of chloroquine treatment.

Genotype Ser413/Ser of PAI-2 polymorphism Ser413/Cys is associated with anti-phospholipid syndrome and systemic lupus erythematosus in a familial case: comparison with healthy controls.

Background: We describe a family with a 7‐year‐old proband case diagnosed with systemic lupus erythematosus (SLE) plus secondary anti‐phospholipid syndrome (APS) as well as two affected paternal aunts. We compared the frequency of these polymorphisms with healthy controls. Objectives: To evaluate the mode of inheritance in this familial case of APS and SLE and the possible association of plasminogen activator inhibitor‐1 (PAI‐1) −675 4G/5G and PAI‐2 Ser413/Cys polymorphisms. To compare the genotype frequency of these polymorphisms with the results found in a Mexican Mestizo population. Methods:PAI‐1 −675 4G/5G and PAI‐2 Ser413/Cys were determined by the polymerase chain reaction restriction fragment length polymorphism (PCR‐RFLP) technique using Bsl I and Mwo I on four generations of the family studied. PAI‐2 Ser413/Cys polymorphism was also determined in 50 healthy individuals of Mexican Mestizo origin. Results: The family pedigree demonstrated that this family did not follow a Mendelian inheritance pattern. When the PAI‐2 Ser413/Cys polymorphism was examined, we found that 60% (3/5) of the relatives homozygous to Ser413/Ser were affected with SLE and/or APS (p = 0.027). The proband case was 4G/5G genotype for the PAI‐1 −675 4G/5G polymorphism. No differences between healthy controls of the Mexican Mestizo population and the family studied for the PAI‐2 Ser413/Cys polymorphism or PAI‐1 −675 4G/5G polymorphisms were found. Conclusions: Our data indicate that this family did not follow the Mendelian inheritance pattern. The Ser413/Ser genotype demonstrated in 60% of the affected members (3/5) of this family might increase the risk for autoimmune syndromes such as APS or SLE.

Circulating TNFRI and TNFRII levels correlated with the disease activity score (DAS28) in rheumatoid arthritis

Objective: To measure levels of soluble tumour necrosis factor alpha (TNFα) receptor type I (sTNFRI) and type II (sTNFRII) in order to correlate them with C-reactive protein (CRP), rheumatoid factor (RF), erythrocyte sedimentation rate (ESR), and disease activity score (DAS28) in RA patients. Methods: We recruited 41 RA patients classified according to American College of Rheumatology (ACR) criteria and 38 healthy subjects (HS). sTNFRI and sTNFRII were measured using an enzyme-linked immunosorbent assay (ELISA) kit. Clinical activity in RA patients was evaluated using the Disease Activity Score using 28 joint counts (DAS28). The statistical analysis was realized using SPSS version 10.0. Results: Soluble TNFRI and TNFRII levels were higher in RA patients (p = 0.04 and 0.001, respectively) than HS. Serum levels of sTNFRI correlated with sTNFRII (r = 0.699, p < 0.0001). sTNFRII correlated with DAS28 (r = 0.375, p = 0.017), RF (r = 0.505, p = 0.004), and ESR (r = 0.323, p = 0.042). Conclusion: The increased levels of both sTNFRI and sTNFRII suggest a secondary event related to the inflammatory state observed in RA, whereas the correlation of sTNFRII with RF, ESR, and DAS28 reflects the preferential TNFRII shedding induced by TNFα. sTNFRII may be useful as an additional inflammatory marker in RA.

The ACTN3 R577X polymorphism is associated with inflammatory myopathies in a Mexican population

Background: The ACTN3 gene encodes the fast muscle protein α-actinin-3. The ACTN3 R577X polymorphism is a premature stop codon and results in absence of α-actinin-3 in 577XX homozygotes. The aim of this study was to determine the ACTN3 genotype in idiopathic inflammatory myopathies (IIMs). Methods: We performed ACTN3 genotyping on 27 patients with dermatomyositis (DM), 10 with polymyositis (PM), and 85 healthy subjects. Muscle enzyme levels of creatine phosphokinase (CPK), lactic dehydrogenase (LDH), aspartate aminotransferase (AST), and alanine aminotransferase (ALT) were recorded at the time of diagnosis and recruitment. Genotyping was performed by polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) and the allele frequency was analysed. Results: A total of 36% of healthy subjects had the ACTN3 577XX polymorphism (α-actinin-3 deficiency), 18% had the 577RR (homozygous wild type) genotype, and 46% 577RX (heterozygous). In DM/PM, 70% had the ACTN3 577XX polymorphism, 6% RR, and 24% RX [odds ratio (OR) 4.12, 95% confidence interval (CI) 1.67–10.33, p < 0.001]. In healthy subjects, the R allele was present in 41% and the X allele in 59% compared to 18% and 82%, respectively, in the IIM group (OR 3.21, 95% CI 1.57–6.66, p < 0.001). Thus, the ACTN3 577X allele seemed to increase the risk of developing IIM, and DM in particular, although this was not related to severity of expression of the phenotype. Conclusions: The ACTN3 577X allele appeared to increase the risk of developing IIM; 70% of IIM patients were deficient in α-actinin-3. By contrast, ACTN3 577XX patients seemed to have less severe disease as reflected in lower muscle enzyme levels.

Evaluation of lipid profile, macular toxicity and clinical manifestations according to APO E genotype in systemic lupus erythematosus and rheumatoid arthritis patients treated with chloroquine

Objective: To investigate the effect of APO E gene polymorphism over lipid profile, macular toxicity and clinical manifestations in RA and SLE patients treated with chloroquine. Materials and methods: We studied 45 RA and 29 SLE patients treated with chloroquine who were classified based on the therapeutic regime of chloroquine into three groups: A) Cumulative dose of 100-300 g, B) > 300 g and C ) Never received chloroquine. Clinical evaluation, fasting lipid profile, visual field testing and stereoscopic photos of the retina were performed. APO E genotype was determined by PCR-RFLP. Results: Reduced apo B levels in RA and SLE according to the cumulative dose of chloroquine 2/3 APO E genotype in a subset of SLE patients were observed. Macular toxicity was independent of both APO E genotype and cumulative chloroquine dose. Conclusions: Reduced apo B levels were observed associated to chloroquine treatment and 2/3 APO E genotype.

Prevalence of antinuclear antibodies in a Huichol population of Mexico.

Antinuclear antibodies have become an important diagnostic aid in systemic rheumatic diseases. Their prevalence in normal individuals has been established to be low (2.3-3.5%) in various studies’, 2. However, it is not clear whether they reflect subclinical disease in all cases, or if they are present only transiently. In our country, we also have found a low prevalence of antinuclear antibodies in normal Mexican mestizos individuals (4.7%)~. Mexican mestizos constitute >95% of the Mexican current population and are the result of the mixture between Spaniards and Mexican Amerindians. With regard to antinuclear antibodies, there are no published studies which deal with the prevalence of these autoantibodies in Indian Mexican groups. For this reason, we studied a group of healthy individuals from a Huichol population for these autoantibodies. The Huichol population is geographically confined to state limits of Nayarit and Jalisco in the western part of Mexico (Figure 1), and had its first European contact in 1531 when Vasco de Quiroga, a Spanish explorer, guided an expedition to that region4. The latest census of Amerindian groups in our country made in 1990, reported a total of 19363 Huicholes (9610 males and 9753 females)5. Based on the known prevalence of these autoantibodies in the normal population, we calculated a representative sample of our study group using a conventional statistical formula. Serum samples were obtained from 46 apparently healthy Huicholes of five small villages: Nueva Colonia, Mesa de Tepic, Las Latas, Tuxpan de Bolanos and San Andres Cohamiata. Sixty three per cent were females (29 of 46) and 37% males (17 of 46). Antinuclear antibodies were detected by indirect immunofluorescence in heat inactivated sera (30 min at 56°C) using Hep-2 cells as substrate (Antibodies Inc., Davis, CA, USA) and a polyvalent detecting reagent conjugated with fluorescein (Antibodies Inc., Davis, CA, USA). Sera were tested at 1:4, 1:16, 1:64 and 1:256 dilutions. Based on our previous experience we considered positive those samples showing a definite fluorescence pattern at 1: 16 dilution or more. Antinuclear antibodies

Coexistence of anti-RNA polymerase III and anti-U1RNP antibodies in patients with systemic lupus erythematosus: two cases without features of scleroderma

Anti-RNA polymerase III (RNAP III) antibodies are highly specific for scleroderma (SSc) and associated with diffuse SSc and renal crisis. Coexistence of anti-RNAP III and other SSc autoantibodies is rarely documented. We report three cases with coexisting anti-RNAP III and anti-U1RNP. Autoantibodies in 3829 sera from rheumatology clinics were screened by immunoprecipitation. Anti-RNAP III-positive sera were also examined by immunofluorescence and anti-RNAP III ELISA. In total, 35 anti-RNAP III-positive sera were identified by immunoprecipitation, in which three had coexisting anti-U1RNP. All three were anti-RNAP III ELISA positive. Two had anti-RNAP I dominant (vs. RNAP III) reactivity and showed strong nucleolar staining. A case with anti-U1/U2RNP (U2RNP dominant) had systemic lupus erythematosus (SLE)–SSc overlap syndrome; however, the remaining two cases had SLE without signs of SSc. All three cases of anti-RNAP III + U1RNP fulfilled ACR SLE criteria but none in the group with anti-RNAP III alone (p = 0.0002). In contrast, only one case in the former group had sclerodermatous skin changes and Raynauds phenomenon, vs. 92% with scleroderma in the latter (p < 0.05). Although anti-RNAP III is highly specific for SSc, cases with coexisting anti-U1RNP are not so uncommon among anti-RNAP III positives (8%, 3/35) and may be SLE without features of SSc.

AB0330 Rheumatoid factor and RO52KDA antibodies are independent predictors of insulin resistance in rheumatoid arthritis

Background The rheumatoid factor (RF) and anti-citrullinated protein antibodies (ACPA) autoantibodies in rheumatoid arthritis (RA), have been used as diagnostic and prognostic tools [1]. However, this traditional perspective has changed toward a major role in RA pathogenesis. Several studies have demonstrated that FR and ACPA autoantibodies positivity beyond its level, might influence disease activity, bone erosions and development of comorbidities. Anti-Ro52kDa antibodies have also been associated with disease severity in RA and might influence the development of comorbidities such as insulin resistance (IR) in RA. Objectives To evaluate the association between RF, ACPA and anti-Ro52 kDa and IR in RA patients. Methods We included 83 RA patients classified according to ACR 1987 and ACR/EULAR 2010 criteria and 90 controls matched for age, gender and body mass index (BMI). Homeostasis Model Assessment-Insulin Resistance (HOMA-IR), anthropometric parameters and antibody positivity (RF, ACPA, Ro52 kDa) were evaluated. Multivariate regressiόn analysis was used to assess the contribution of autoantibodies, adiposity and disease activity to insulin resistance in RA. Results Patients positive for RF or anti-Ro52 kDa showed higher levels of basal insulin (P=0.009, P=0.006) and HOMA-IR. DAS-28 ESR was correlated with basal insulin (r=0.31, P=0.01) and HOMA-IR (r=0.29, P=0.02). We also observed positive correlations between serum triglycerides (r=0.47, P=0.01) and HDL-c (r=-0.38, P=0.02) and basal insulin. Multivariate analysis showed that Triglycerides, HDL-c, DAS-28, RF and anti-Ro52 kDa were independent predictors of basal insulin and HOMA-IR in patients with RA. Conclusions In RA, RF or anti-Ro52 kDa are independent predictors of IR. This phenomenon might be linked to the network of inflammation, adipokine secretion, since disease activity was also precitive of higher basal insulin. Both RF and anti Ro52 kDa, along with disease activity are independent predictors of IR in RA patients without comorbidities. References Watad A, Amital H: ACPAs Are Much More Than Diagnostic Autoantibodies. Rambam Maimonides Medical Journal 2016, 7(4). England BR, Thiele GM, Mikuls TR: Anticitrullinated protein antibodies: origin and role in the pathogenesis of rheumatoid arthritis. Current Opinion in Rheumatology 2016. Schuntermann MF: [References for evaluation scales in quality assurance in rehabilitation–1. I. Scales for determining adverse sequelae of illnesses–an introduction]. Die Rehabilitation 1995, 34(1):I-III. Matsudaira R, Tamura N, Sekiya F, Ogasawara M, Yamanaka K, Takasaki Y: Anti-Ro/SSA antibodies are an independent factor associated with an insufficient response to tumor necrosis factor inhibitors in patients with rheumatoid arthritis. Journal of Rheumatology 2011, 38(11):2346–2354. Disclosure of Interest None declared

SAT0374 The euromyositis registry: an international description of myositis

Background The idiopathic inflammatory myopathies (IIM) represent a rare and heterogeneous group of multisystem autoimmune diseases. The rarity of IIM has hampered research efforts, resulting in remarkably limited therapeutic evidence. The EuroMyositis Registry was created to pool resources and expertise across the international IIM research community. Objectives To describe the phenotypic characteristics of different IIM subtypes. Associations with malignancy, interstitial lung disease (ILD), cardiac involvement and dysphagia were assessed. Methods Pooled data from the EuroMyositis Registry (from Belgium, China, Czech Republic, Hungary, Italy, Mexico, Norway, Sweden, Switzerland, UK, Vietnam) were obtained. Associations were assessed using logistic regression and multinomial logistic regression with polymyositis (PM) as the reference group. Results Data regarding 3,196 patients were analysed. The UK was the largest contributor (n=1,307). The most common diagnoses were dermatomyositis (34%), PM (32%) and connective tissue disease (CTD)-overlap myositis (15%). In those with anti-synthetase syndrome (7%), 85% had muscle weakness, 86% ILD and 58% arthritis. Overall, 43% had a myositis specific antibody, most commonly anti-Jo1 autoantibodies (20%). Glucocorticoid usage was noted in 98%. Most commonly used disease modifying agents were methotrexate (71%) and azathioprine (50%). Malignancy occurred in 9% and was associated with a diagnosis of dermatomyositis (Relative Risk Ratio [RRR] 1.68, 95% CI 1.09–2.56, p=0.018). Cardiac involvement occurred in 9%, most commonly in those with CTD-overlap myositis (13%), and was associated with a higher Health Assessment Questionnaire disability index (1 versus 0.75, OR 1.40, 95% CI 1.03–1.91, p=0.031). Dysphagia occurred in 39% and was associated with a diagnosis of CTD-overlap myositis (RRR 2.25, 95% CI 1.61–3.15, p<0.001). Conclusions This large international cohort demonstrates the heterogeneity of IIM and the burden of associated malignancy, ILD, cardiac and gastrointestinal involvement. The EuroMyositis Registry facilitates international collaborative research outputs, underlining the benefits of harmonised data collection methodology between centres. Acknowledgements This work was supported by researchers at the National Institute for Health Research (NIHR) Biomedical Research Unit. The views expressed are those of the authors and not necessarily those of the UK National Health Service, the NIHR or the UK Department of Health Disclosure of Interest J. Lilleker: None declared, J. Vencovsky: None declared, G. Wang: None declared, L. Wedderburn Grant/research support from: The UK JDM Cohort and Biomarker study is supported by grants from the NIHR and Myositis UK, L. Diederichsen: None declared, J. Schmidt: None declared, P. Jordan: None declared, O. Benveniste: None declared, M. G. Danieli: None declared, K. Dankό: None declared, N. T. P. Thuy: None declared, M. Vazquez-Del Mercado: None declared, Ø. Molberg: None declared, B. De Paepe: None declared, J. De Bleecker: None declared, B. Maurer Grant/research support from: AbbVie, Protagen, EMDO, Novartis, Roche, Actelion, N. Pipitone Speakers bureau: GRAPPA Workshop, Alfa-Wassermann, N. McHugh: None declared, Z. Betteridge: None declared, P. New: None declared, R. Cooper: None declared, W. Ollier: None declared, J. Lamb Grant/research support from: MedImmune, N. S. Krogh: None declared, I. Lundberg Grant/research support from: Astra-Zeneca, Bristol-Myers Squibb, Consultant for: Bristol-Myers Squibb, Idera, H. Chinoy Grant/research support from: MedImmune, Novartis