No abstract

Background Diagnosis of antiphospholipid syndrome (APS) still remains a laboratory challenge due to the great diversity of antiphospholipid antibodies (aPL) and their significance regarding APS classification criteria. Standardized enzyme-linked immunosorbent assay (ELISA) for antibodies to cardiolipin (aCL) and b2-glycoprotein I (ab2GPI) has been indispensable in the diagnosis and management of APS. Multiplex detection of aPL profile seems to be a new approach for risk assessment in APS. Objectives The comparison of the novel multi-line dot assay (MLDA) with results of the routine ELISA technique for detection of aPL in APS. Methods We studied 36 patients (pts) with APS (5 men, 31 women, median age 34, 29-44 years). Diagnosis of APS had been established according to the 2006 international classification criteria. 15 (41,7%) APS pts met the diagnostic criteria for systemic lupus erythematosus (SLE), 28 (77,8%) had a history of arterial and/or venous thrombosis, 21 (58,3%) suffered from fetal loss. As a disease control group, 14 pts with SLE without APS (3 men, 11 women, median age 35, 27-45 years) were included. IgG/IgM antibodies to CL, b2GPI, phosphatidylserine (aPS) and phosphatidylinositol (aPI) in the pts sera were detected using a commercial available MLDA (“Medipan”, Germany). For comparison, the serum levels of IgG/IgM aCL and ab2GPI were measured by ELISA kit (“Orgentec”, Germany). Results APS pts demonstrated a significantly higher frequency of IgG aCL, IgG and IgM ab2GPI in ELISA and IgG aCL in MLDA compared to SLE pts (p<0,05) (table 1). The agreement between both methods was good for IgG aCL, IgM aCL and fair for IgG ab2GPI, IgM ab2GPI (kappa=0,620, 0,628, 0,380 and 0,388, respectively). The frequency of discrepant results for IgG aCL, IgM aCL, IgG/ IgM ab2GPI was 16%, 18% and 30%. Multiple positivity for different aPL detected by ELISA occurred significantly more frequently in APS pts (27,8%) in contrast to SLE pts (7,1%, p<0,05). The number of multiple positive samples evaluated by MLDA in the APS group (13,9%) did not demonstrated a significant higher prevalence in comparison with the control group (7,1%, p>0,05). There was a relationship between ischemic stroke and the combination of high positive levels of IgG/IgM aCL, ab2GPI, aPS and aPI detected by MLDA in APS pts (OR 95%CI 2,45, 1,09-5,55, ρ=0,049). We didn’t find any associations between subtypes of aPL and thrombosis localization. Image/graph Conclusions MDLA present an alternative to ELISA for aPL evaluation and profiling in APS. The rate of false-positive aPL detected by MLDA in SLE pts was higher compared to ELISA. The combination of four high positive aPL measured by MLDA is a risk factor of ischemic stroke in APS. Disclosure of Interest None Declared

Systemic lupus erythematosus (SLE) - chronic systemic disease, which associates with genetic defects of immunoregulatory processes and leads to uncontrolled production of antibodies to native cells and its elements with autoimmune inflammation development. There is a case report of human intravenous immunoglobulin use in lupus patient with the multiple resistances to antirheumatic drugs and many adverse reactions of the therapy.

Background The presence both of antiphospholipid antibodies (aPL) and anti-dsDNA is immunological features of diagnostic criteria of SLE. The level of anti-dsDNA usually reflects renal involvement of SLE and antiphospholipid antibodies (aPL) is associated with thrombotic events. Cross-reaction between these antibodies is discussed. Objectives To assess the frequency of anti-dsDNA, increasing of CIC, hypocomplementemie in SLE and APS (in primary and secondary) and their connection with anticardiolipin antibodies (aCL) and clinical manifestations of the disease. Methods We studied 365 patients in mean age 31,9±10,9 years (from 14 to 63yrs.) and disease duration 9,1±7,5years (from6 month to 36 yrs). SLE was diagnosed in 135 (56M, 79F) patients (ACR criteria, 1997) and 230 (55M, 175F) patients had APS. Secondary APS (SLE+APS) was in 156 of 230 patients and primary – in 74. Anti-dsDNA and aCL were measured by ELISA. CH50 was evaluated by 50% hemolytic activity. C3, C4 were determined by immunonephelometric assay, CIC – by turbidimetric assay with 3,5% PEG. Results High level of anti-dsDNA was found to have 80% of SLE patients (mean level 68,5±28,6 OD from 30 to 120) and it was associated with glomerulonephritis and hipocomplementemia. Increased levels of IgG-aCL were detected in 22% of SLE patients and IgM-aCL - in17%, but nobody of them had any APS features. There was correlation between IgG-aCL and anti-dsDNA, CIC and IgG. High level of anti-dsDNA was found in 75% of SLE+APS patients and it was correlated with activity disease estimating by SLEDAI score (35.2±14.2 vs 12.3±5.7) but not with renal involvement. Fifty six percent of 156 SLE+APS patients were IgG-aCL positive and 28% - IgM-aCL positive. Thrombosis were revealed in 48% of IgG-aCL positive SLE+APS patients and thrombocytopenia in 15% of IgM-aCL positive patients. Immunological abnormalities have revealed in 25% of 74 PAPS patients: 25% had false positive test for siphylis, 61% - high level of CIC, 7% -anti-DNA and 19% - hipocomplementemia. High level of anti-dsDNA in PAPS patients was associated with microvascular complication: digital necrosis and Rayunad’s phenomen. There was correlation between IgG-aCL and CIC in APS patients. Conclusions High level of anti-dsDNA in SLE patients associated with renal damage. There was correlation between the levels of anti-dsDNA with SLEDAI score in SLE+APS patients and wasn’t associated with renal involvement. Immunological abnormalities (CIC, low C3, C4) except aCL-positivity have revealed in PAPS patients. Disclosure of Interest None Declared

Background Last years the reasons of earlier and more frequent atherosclerosis and cardiovascular/cerebrovascular disorders in rheumatic diseases and also the participation of antiphospholipid antibodies (aPL) in atherothrombosis are up for debate. Analysis of blood concentration of deferent autoantibodies, acute-phase reactants and lipid profile as well as investigation of traditional and nontraditional risk factors and intima-media thickness in patient (pts) with antiphospholipid syndrome (APS) is topical for the understanding of the role of atherosclerosis in APS. Objectives To investigate levels of high sensitive CRP (hs-CRP), interleukin 6 (IL 6), tumor necrosis factor α (TNFα), soluble TNFα-receptor 1 (sTNFα-R1) and intercellular adhesion molecule 1 (ICAM-1) in APS pts. Methods The total of 96 APS pts (52 with primary APS (PAPS) and 44 with SLE+APS) and 29 age– and sex–matched healthy controls (HC) were included. Thromboses in past history were registered in 83/96 (86%) pts: in 21 – arterial (AT), in 38 – venous (VT), in 24 – AT+VT. Serum markers of inflammation and endothelial cells activation were measured: IL 6 – in 89 pts and 20 HC, TNFα – in 73 pts and 20 HC, sTNFα-R1 – in 90 pts and 28 HC and ICAM-1 – in 66 pts and 20 HC, respectively. All the pts underwent electrocardiography, echocardiography, Holter monitoring, ultrasonography and laboratory testing including aPL, hs-CRP and lipid profile of plasma. Results Serum levels of hs-CRP, IL 6, TNFα, sTNFα-R1 and ICAM-1 were significantly higher in pts compared to HC (p<0.05 in all cases). Levels of IL 6 and sTNFα-R1 in PAPS pts were lower than in SLE+APS pts (1.01 [0.27; 1.90] pg/ml; 2.43 [1.05; 4.13] ng/ml vs 2.30 [1.90; 3.30] pg/ml; 3.89 [2.20; 6.00] ng/ml respectively, p<0.05). Concentration of ICAM-1 was significantly higher in pts with VT than in pts with AT, with AT+VT and without T (445.2 [342.7; 426.4] vs 316.0 [282.3; 349.4] vs 356.7 [327.2; 383.4] vs 344.1 [314.2; 379.2] ng/ml, respectively; p=0.04). Coronary heart disease (CHD) positively associated with increased levels of sTNFα-R1: CHD was diagnosed in 17/45 pts with elevated sTNFα-R1 levels and in 3/45 pts with normal sTNFα-R1 levels (OR 2.13 [95%IC 1.51; 2.99], p<0.001). During the first year after thrombosis there was an inverse association between postthrombotic time duration (PTTD) and IL 6 and TNFα: increased levels of IL 6 and TNFα were detected in 4/4 (100%) and 8/15 (53%) pts with acute thrombosis (PTTD ≤1 week), in 15/16 (94%) and 5/5 (100%) pts with 1<6 months and in 12/12 (100%) and 19/21 (90%) pts with 6<12, respectively, p<0.05 in all cases. Levels of IL 6 (3.36 [2.33; 4.13] pg/ml) and TNFα (5.75 [3.19; 8.28] ng/ml) in pts with PTTD ≤1 week were higher than in pts with 1<6 months (IL 6 =1.88 [1.14; 4.08] ng/ml and TNFα =4.53 [3.55; 6.81] ng/ml) and 6<12 months (IL 6 =1.72 [1.01; 2.86] pg/ml and TNFα =2.9 [1.96; 3.49] ng/ml), p<0.05 in all cases. Conclusions Serum levels of hsC-RP, IL 6, TNFα, sTNFα-R1 and ICAM-1 were significantly elevated in pts compared to HC. ICAM-1 ...