Bt Martín-Márquez

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.

The −174G/C and −572G/C Interleukin 6 Promoter Gene Polymorphisms in Mexican Patients with Rheumatoid Arthritis: A Case-Control Study

Objective. There is a lack of information about the genotype frequencies of IL-6 −174G/C and −572G/C polymorphisms in Mexicans with rheumatoid arthritis (RA). Therefore, the aim of this study was to evaluate the association of the IL-6 −174G/C and −572G/C polymorphisms in Mexican mestizo with RA. Methods. We included 137 patients with RA and 102 healthy controls. Patients were assessed for clinical characteristics. IL-6 −174G/C and −572G/C polymorphisms were genotyped using PCR-RFLP analysis. Allele and genotype frequencies and the Hardy-Weinberg equilibrium were computed. Odds ratios (ORs) were computed to identify the risk for RA associated with the presence of GG genotype in comparison with the GC or CC genotypes. Results. The genotype −174GG occurred at a higher frequency in cases and controls (77.4% versus 78.4%, P = 0.845). We found similar results for the genotype −572GG (54% in patients versus 60.8% in controls, P = 0.295). Conclusions. This is the first study to evaluate the association of −174G/C and −572G/C polymorphisms of the IL-6 gene with RA in Mexican mestizo patients. These two polymorphisms were not associated with RA in the studied sample. Additional studies are required to evaluate if these IL-6 polymorphisms have relevance to the development of more severe disease.

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.

Polymorphism rs2073618 of the TNFRSF11B (OPG) Gene and Bone Mineral Density in Mexican Women with Rheumatoid Arthritis

Osteoporosis (OP) is highly prevalent in rheumatoid arthritis (RA) and is influenced by genetic factors. Single-nucleotide polymorphism (SNP) rs2073618 in the TNFRSF11B osteoprotegerin (OPG) gene has been related to postmenopausal OP although, to date, no information has been described concerning whether this polymorphism is implied in abnormalities of bone mineral density (BMD) in RA. We evaluated, in a case-control study performed in Mexican-Mestizo women with RA, whether SNP rs2073618 in the TNFRSF11B gene is associated with a decrease in BMD. RA patients were classified as follows: (1) low BMD and (2) normal BMD. All patients were genotyped for the rs2073618 polymorphism by PCR-RFLP. The frequency of low BMD was 74.4%. Higher age was observed in RA with low BMD versus normal BMD (62 and 54 years, resp.; p < 0.001). Worse functioning and lower BMI were observed in RA with low BMD (p = 0.003 and p = 0.002, resp.). We found similar genotype frequencies in RA with low BMD versus RA with normal BMD (GG genotype 71% versus 64.4%, GC 26% versus 33%, and CC 3% versus 2.2%, resp.; p = 0.6). We concluded that in Mexican-Mestizo female patients with RA, the rs2073618 polymorphism of the TNRFS11B gene is not associated with low BMD.

AB0936 Tuberculosis Infection in Mexican Patients with Idiopathic Inflammatory Myopathies: Report of three Cases and Review of the Literature

Background Tuberculosis (TB) is a public health problem, a disease originated by M. tuberculosis (MTB) and other members of Mycobacterium tuberculosis complex. There is scarce information about the MTB infection in polymyositis/dermatomyositis (PM/DM) patients. Objectives To describe the prevalence and clinical presentation of TB infection in patients diagnosed as PM/DM from the two main cities of Mexico: Guadalajara and Mexico City. Methods A group of 196 patients with IIM recruited from 2009 to 2014 from four public hospitals were included. We described the clinical characteristics and the possible risk factors for the development of TB infection. Results We identified three cases of extrapulmonary MTB infection mainly TB tenosynovitis (2) and meningeal TB (1) from a cohort of 196 patients with idiopathic inflammatory myopathies (IIM) recruited in a period of 5 years (2009 to 2014). The three cases presented the TB infection around 12 weeks of immunosuppressive therapy. Conclusions Diagnosis of extrapulmonary TB should be considered in the clinical approach of IIM patients especially in Mexico and countries with high prevalence of TB infection. Previous to immunosuppressive therapy in IIM, we suggest to do screening for MTB using Tuberculin Skin Test and chest X-ray. In cases of high clinical suspicion of TB infection, we might use Quantiferon™ (Cellestis Limited, Melbourne, Australia) and Xpert MTB-RIF™ (Cepheid, Sunnyvale, California, US). Disclosure of Interest None declared

FRI0081 Titers of Anti-Ccp Antibodies, IL-6, Tnf-Alpha and CRP Are Associated with Increased Carotid Intima-Media Thickness: Cross-Sectional Analysis of A Rheumatoid Arthritis Group without Cardiovascular Risk Factors

Background The main causes of death in Rheumatoid Arthritis (RA) are cardiovascular events related to early atherosclerosis, myocardial infarction and congestive heart failure. The risk for cardiovascular disease it it not completely explained by the traditionally accepted cardiovascular risk factors. Inflammatory pathways of RA might be involved in the pathogenesis of subclinical atherosclerosis. Objectives To evaluate the relationship of anti-CCP antibody titers, serum IL-6, TNFa and high sensitivity c-Reactive Protein with increased carotid intima-media thickness (cIMT) in patients with RA without cardiovascular risk factors. Methods Sixty patients with RA, 27 anti-CCP positive and 33 anti-CCP negative, and 62 controls (HC) matched by age and gender. All groups were assessed for comorbidities, demographics, clinical evaluations, and cardiovascular risk calculated by atherogenic index of plasma (AIP). The cIMT was performed by high-resolution B-mode ultrasound by a single operator. Serum titers of anti-CCP, hs-cRP, levels of TNFα and IL-6 were measured by ELISA. Results The anti-CCP positive RA patients showed increased cIMT compared to HC and anti-CCP negative patients (p<0.001). Anti-CCP positive vs. Anti-CCP negative patients, had increased disease activity (DAS28) (p=0.05), increased AIP (p<0.001), higher TNFα (20.54±5.38 vs 32.77±12.30, p=0.002) and IL-6 (18.54±14.09 vs 65.07±34.95, p<0.001). The cIMT correlated with titers of anti-CCP (r=0.539, p<0.001), TNFα (r=0.791, p<0.001) and IL-6 (p=0.794, p<0.001). In multiple regression analysis, cIMT was associated with hs-cRP (p=0.05) and anti-CCP levels (p=0.006). Conclusions High levels of anti-CCP and hs-cRP are useful markers of an increased cIMT and cardiovascular risk supporting a clinical role of the preventive measurement of cIMT in RA Disclosure of Interest None declared DOI 10.1136/annrheumdis-2014-eular.5947

SAT0135 Low Levels of CD36 in Peripheral Blood Monocytes in Subclinical Atherosclerosis in Rheumatoid Arthritis: A Cross-Sectional Study of A Mexican Population

Background Rheumatoid arthritis (RA) patients have a higher risk for atherosclerosis mainly as a consequence of chronic systemic inflammation, notwithstanding, the implicated pathways have not been completely understood. CD36 is a scavenger receptor relevant for atherogenesis, there are still controversies about its role in this process and there is no clinical information about CD36 and subclinical atherosclerosis in human patients with RA. Objectives To Evaluate the relation between the membrane expression of CD36 in peripheral blood monocyte cells and carotid intima media thickness (cIMT) in patients with RA. Methods We included 37 patients with RA (ACR 1987) attending the rheumatology department of Hospital Civil “Dr. Juan I. Menchaca” Guadalajara, Jalisco, Mexico. The cIMT was assessed by high definition mode B ultrasound and patients were grouped as increased cIMT if >0.6 mm or as no increased cIMT if <0.6 mm. Disease activity was evaluated with the DAS28-cRP. Total cholesterol (TC), triglycerides (Tg), high density lipoprotein cholesterol (HDL-c) and low density lipoprotein cholesterol (LDL-c) were measured. The atherogenic index of plasma (AIP) was calculated. Anti-CCP antibodies, serum IL-6 and TNFa were measured by ELISA. Membrane expression of CD36 was measured by immunofluorescence flow cytometry in PMBC. Data was analyzed with the software WinMDI 2.9 and SPSS v. 22 IBM, Inc. Results Of the studied patients, 24 (65%) had increased cIMT, this group showed higher serum TC (p<0.001), Tg (p=0.006), oxLDL (p<0.001) and AIP (p<0.001) and lower serum HDL-c (p=0.005). Titers of hs-cRP, TNFα, IL-6 and anti-CCP also were higher in the increased cIMT group. Mean fluorescence intensity (MFI) for CD36 was lower in the increased cIMT compared with the no increased cIMT (46.84±40.46 vs. 122.70±34.71, p<0.001). The cIMT was negatively correlated with CD36 MFI. We observed negative correlations between CD36 and TNFa (-0.729, p<0.001) and IL-6 (-0.822, p<0.001). Conclusions Low membrane expression of CD36 is observed in RA patients with subclinical atherosclerosis, Further, studies are needed to validate our findings and clarify the regulation and role CD36 in subclinical atherosclerosis in RA Disclosure of Interest None declared DOI 10.1136/annrheumdis-2014-eular.6032

FRI0321 Efficiency of interferon-Γ release assay for screening for latent tuberculosis in patients with systemic lupus erythematosus

Background Leading causes of death in systemic lupus erythematosus (SLE) are infections, kidney failure and cardiovascular disease. Although most infections are bacterial, there is a greater prevalence of tuberculosis (TB) secondary to multiple immunological abnormalities and immunosuppression. The tuberculin skin test (TST) and chest radiography are not sufficient to detect latent TB infection (LTBI) in this group of patients. Currently interferon-Yrelease assay (IGRAS): Quantiferon-TB Gold In-Tube ™ (QFT-GIT) and T-SPOT.TB ™ (T-Spot), have shown greater superiority to diagnose LTBI in immunocompetent patients but efficiency is unknown in immunosuppressed patients with SLE. Objectives The objective of the present study is to evaluate the efficiency of IGRAS in screening for LTBI in patients with SLE. Methods We performed a cross-sectional study that included patients with SLE according to the ACR criteria, ≥ 16 years old and signed informed consent; pregnant patients were not included, nor with active TB or with antituberculosis treatment. Patients underwent a questionnaire to obtain epidemiological and clinical data of SLE and TB, and TST and IGRAS were performed. The statistical analysis included descriptive arithmetic, chi square for categorical variables, Student’s t test for quantitative variables and kappa for concordance. Results We reviewed a total of 106 patients with SLE, with a mean age of 34.7 ± 13.2 and 95% were female. A history of BCG was found in 90% and 84% presented a scar. Only 8% reported contact with TB patients and 12% were employees or residents of health or correctional institutions. A previous PPD was reported in 8%, of which 11% had a positive result. The 4% with previous diagnosis of TB had received treatment, 75% with lung and 25% with kidney involvement. Comorbidities: 28.3% with hypertension, 2.8% diabetes mellitus and 16% with dyslipidemia. In the present study 9% had a positive PPD, with an average of 5.53 ± 1.92 mm. The QFT-GIT test reported 14% positive, 10% indeterminate and 76% negative. The latter test average was 1.77 ± 0.68 IU / ml in general, 3.14 ± 2.85 in the group of patients with a positive test, 1.41 ± 3.04 in the indeterminate and 0.11 ± 0.11 in the negative group. A correlation between positive results of the two tests was 0.24. Conclusions In our group of patients with SLE a significant number of positive tests was observed for the diagnosis of LTBI, finding a fair concordance between the two tests, but a large number of indeterminate tests were seen. Further studies are needed to determine the usefulness of these tests in SLE and the need for a lower cutoff of IGRAS in this type of patient. References: Centers for Disease Control and Prevention. Updated Guidelines for Using Interferon Gamma Release Assays to Detect Mycobacterium tuberculosis Infection — United States, 2010. MMWR 2010; 59: 1-26 Prabu V, Agrawal S. Systemic lupus erythematosus and tuberculosis: A review of complex interactions of complicated diseases. J Postgrad Med 2010; 56: 244-50 Disclosure of Interest: None Declared

Placental cytokine expression and NF‐κB activity in women with rheumatic diseases

Exacerbations occur during pregnancy in women with systemic lupus erythematosus (SLE) in up to 60% of the cases (1). In pregnant women with rheumatoid arthritis (RA), more than 70% experience improvement of symptoms (2). The balance in the T helper (Th)1/Th2 cytokine profile is an unsolved issue in autoimmune rheumatic diseases, particularly in pregnancy. We investigated the expression of the Th1/Th2 cytokine profile and NF-kB activity on placental tissue from SLE and RA pregnant patients compared with healthy pregnant women (HPW). Pre-existing clinical conditions, such as diabetes, hypertension, and other overlapping diseases were considered as exclusion criteria. Clinical evaluation of SLE and RA patients was performed using the MEX-SLEDAI (3) or the EULAR modified count score (4), respectively. A homogeneous group of women was studied, with a mean age of 27 years and a mean of 5 years evolution at diagnosis. Twenty-six placental specimens were collected: nine SLE, seven RA, seven HPW, one primary anti-phospholipid syndrome (APS), one inflammatory myopathy, and one haemosiderosis plus arthropathy. All SLE and two RA patients were being treated with low-dose prednisone (v10 mg/day). All except one of the patients had successful pregnancies at term (one premature birth). Cytokine expression from placental tissue homogenates was determined by semiquantitative reverse transcription polymerase chain reaction (RT-PCR) (5) and enzyme-linked immunosorbent assay (ELISA). The expression of mRNA from placental tissue is shown in Table 1. All cytokines evaluated have non-significant differences between groups. However, it is important to note that interleukin (IL)-1b expression was the predominant cytokine in all groups studied except for RA. In addition, SLE placental tissue homogenates showed a predominant mRNA IL-4 expression followed by IL-10 (Th2 pattern). The rest of the samples tested for dermatomyositis/polymyositis (DM/PM), APS and haemosiderosis expressed only pro-inflammatory cytokines IL-1b and tumour necrosis factor-a (TNF-a). In all groups the predominant cytokine expressed was IL1b (mean 37.5¡30.8 pg/mL, range 7.65–133.7 pg/ mL). DNA binding activity of NF-kB determined by electrophoretic mobility shift assay (EMSA) (6) was present in two RA patients, two HPW, and one SLE patient as shown in Figure 1. All analyses were performed using SPSS 10.0 software. Probability values lower than 0.05 were considered statistically significant. The balance between the Th1/Th2 cytokine profile is an unsolved issue in autoimmune rheumatic diseases, particularly in the presence of