Lidia Ostanek

The expansion of CD4+CD28- T cells in patients with rheumatoid arthritis.

Clonal expansion of CD4+CD28- T cells is a characteristic finding in patients with rheumatoid arthritis (RA). Expanded CD4+ clonotypes are present in the peripheral blood, infiltrate into the joints, and persist for years. CD4+CD28- T cells are oligoclonal lymphocytes that are rare in healthy individuals but are found in high percentages in patients with chronic inflammatory diseases. The size of the peripheral blood CD4+CD28- T-cell compartment was determined in 42 patients with RA and 24 healthy subjects by two-color FACS analysis. The frequency of CD4+CD28- T cells was significantly higher in RA patients than in healthy subjects. Additionally, the number of these cells was significantly higher in patients with extra-articular manifestations and advanced joint destruction than in patients with limited joint manifestations. The results suggest that the frequency of CD4+CD28- T cells may be a marker correlating with extra-articular manifestations and joint involvement.

Therapy with infliximab decreases the CD4+CD28- T cell compartment in peripheral blood in patients with rheumatoid arthritis

Chronic inflammatory syndromes such as rheumatoid arthritis (RA) are associated with high frequencies of CD4+CD28− T cells. The number of these cells is genetically determined and may also be a consequence of chronic exposure to tumor necrosis factor-alpha (TNFα). The aim of this study was to examine whether the reported efficacy of anti-TNFα therapy in RA involves a resurgence of T cell populations that re-express CD28. After 36-week therapy with infliximab, a significant decrease in CD4+CD28− T cells in RA patients was observed in comparison with baseline. The results suggest that TNFα-neutralizing therapy may restore T cell homeostasis and reduce expansion of the CD28− T cells, which are cytotoxic and may contribute to organ manifestations in RA.

The distribution of human endogenous retrovirus K-113 in health and autoimmune diseases in Poland

OBJECTIVE During the evolution of the human genome, a number of retroviral integrations have occurred creating a group of human endogenous retroviruses (HERVs). As of now several studies have pointed to the association of HERVs with certain autoimmune diseases such as RA, SLE, multiple sclerosis (MS) and SS as well as various neoplasms. In this study, we investigated the prevalence of HERV-K113 in patients with RA, SLE and in healthy subjects in the Polish population. METHODS Genomic DNA samples from 155 RA patients, 139 SLE patients and 261 newborns (as controls) were tested for the presence of the HERV-K113 allele using PCR. Each individuals DNA was genotyped for null, homozygous or heterozygous insertion of HERV-K113. RESULTS Our data revealed statistically significant differences in the insertion frequencies of HERV-K113 between the groups of RA and SLE patients vs healthy controls (provirus DNA was found in 14.19, 15.11 and 8.05% of individuals, respectively). No homozygous individuals for the K113 allele were found in each of the groups. There was no evidence for HERV-K113 association with clinical features in either group. CONCLUSION Our study-the first such performed for the Polish population-provides a consistent observation with previous reports on the genetic association of HERV-K113 integrations in autoimmune disorders. Here, we found that the prevalence of insertionally polymorphic HERV-K113 was significantly increased in Polish patients with SLE and RA.

The urinary excretion of pyridinoline and deoxypyridinoline during rheumatoid arthritis therapy with infliximab

Rheumatoid arthritis is a systemic disease that causes inflammation and joint destruction. As a result of pathological destruction in bone and cartilage, crosslinks in collagen are resorbed more rapidly. This causes a rise in circulating collagen crosslink levels and their urinary excretion. In RA, apart from the crosslink resorption at the site of inflamed joints, there may be increased resorption due to general bone loss associated with disease activity. The aim of this study was to evaluate the influence of therapy with infliximab on urinary excretion of pyridinoline (PYD) and deoxypyridinoline (DPYR) as a markers of collagen degradation and its correlation with clinical and biochemical parameters of disease activity. Seventeen patients with active rheumatoid arthritis treated with infliximab were recruited into the study. The therapy resulted in the reduction in the symptoms of RA and urinary excretion of PYD and DPYR. The urinary excretion of PYD correlated with a number of swollen joints, morning stiffness, CRP and ESR. The urinary excretion of DPYR correlated during infliximab therapy with the number of swollen and tender joints and morning stiffness. The measurement of urinary excretion of PYR and DPYR may give insight into bone metabolism and help us to better understand the actual changes in bone and cartilage caused by RA and its treatment.

High prevalence of anti-beta-2 glycoprotein-I and anti-prothrombin antibodies in adult-onset Still’s disease. Comment on “Portal vein thrombosis in adult-onset Still’s disease: a case report and literature review”

We have followed with great interest case report by Morita et al [1]. In our group of eight patients diagnosed with Adult-onset Still’s disease (AOSD) according to Yamaguchi criteria, we reviewed presence of antiphospholipid antibodies. They were detected in six of eight patients: the most common were anti-prothrombin (anti-PT) followed by anti-beta2 glycoprotein-I antibodies (anti-b2GPI) (Table 1). D-dimer level was elevated in all patients; however, no related thrombosis was diagnosed. Only one patient presented with deep vein thrombosis at the time of AOSD relapse, and one woman had a history of miscarriage, however, not fulfilling current antiphospholipid syndrome classification criteria. Our first reflection was how difficult differential diagnosis of liver disease in AOSD is, as raised serum aminotransferase (AT) level may be associated with high

Chronic Urticaria and Mild Arthritis Associated with Autoimmune Thyroid Disease: Successful Treatment with L-Thyroxine

Sir, Some cases of chronic urticaria, regarded so far as idiopathic, are in fact of autoimmune origin (1). Autoimmune thyroid disease (ATD) is characterized by the presence of anti-thyroid antibodies. Association of chronic urticaria and ATD is well known. Most studies have found a 15–30% prevalence of anti-thyroid antibodies in patients with chronic urticaria (2, 3). There have been some reports of successful treatment with L-thyroxine of either arthritis (4, 5) or autoimmune urticaria (1, 3) associated with ATD, as well as in euthyroid patients (3–5).

03.03 Serum interleukin 23 in polish patients with systemic lupus erythematosus – association with obesity and peripheral vascular disease

Background Cytokine-mediated immunity plays a crucial role in the pathogenesis of various autoimmune diseases including systemic lupus erythematosus (SLE). The aim of this study was to evaluate association between serum levels of IL-23 and vascular involvement in SLE patients. Materials and methods Study was performed in 94 SLE patients (82 women and 12 men) aged 19–73 years and in 27 age and gender matched controls. Serum IL-23 was measured with ELISA method using R and D Systems tests. Carotid intima-media thickness and the presence of atherosclerotic plaques in carotid and lower extremities arteries were analysed with B-mode ultrasound. Ankle-brachial and high resistance indexes were measured with Doppler ultrasonography. We took into account classical cardiovascular risk factors (hypertension, dyslipidemia, hyperglycemia, overweight/obesity, smoking, oral contraceptives, positive family history of cardiovascular disease), selected clinical manifestations (cardiovascular, cerebrovascular, lupus nephritis, Raynaud’s phenomenon, livedo reticularis, vasculitis, other thromboembolic complications), profile of autoantibodies (antinuclear, antiphospholipid, anti-neutrophil cytoplasmic, anti-endothelial cell). Statistical analysis was performed with: chi 2Yates, chi 2Pearson, rank Spearman correlations tests, logistic regression analysis and multivariate stepwise analysis. Results Concentrations of IL-23 significantly differed between SLE patients and the controls (p=0.0005). Patients with high levels of IL-23 more frequently developed atherosclerosis showed as the presence of plaques in right common femoral artery (OR=10.1; 95% CI:1.2–85.1) and lupus nephritis (OR=3.2; 95% CI:1.1–9.6). Among classical atherosclerotic risk factors only obesity was significantly associated with IL-23 (OR=3.8; 95% CI:1.2–12.3). Immunological characteristics significantly related to IL-23 were anti-phosphatidylethanolamine antibodies (OR=12.7; 95% CI:1.5–108.1) and anti-SS-B antibodies (OR=11.8; 95% CI:1.5–94.8). Association with anti-cardiolipin and anti-prothrombin antibodies was on the border of statistical significance (OR=2.3; 95% CI:0.9–5.7 and OR=8.4; 95% CI:1.0–71.1 respectively). Conclusions IL-23 may be involved in lupus nephritis pathogenesis. 2. IL-23 through its significant association with obesity and antiphospholipid antibodies may promote hypercoagulable state contributing to atherothrombosis development in SLE patients.

A5.07 The role of immunologic and inflammatory factors in the risk of microvascular and macrovascular impairment development in systemic lupus erythematosus – preliminary data

Background and objectives Vascular disorders are a well recognised clinical problem in systemic lupus erythematosus (SLE). This preliminary study was designed to evaluate the association between cerebral circulation changes, carotid arteries involvement as well as nailfold capillaroscopy (NC) abnormalities and immunologic/inflammatory markers, classical atherosclerosis risk factors and organ involvement in SLEpatients. Materials and methods The study was performed in 30 SLE patients. Bilateral transcranial doppler (TCD) monitoring over the middle cerebral arteries according to the criteria of the International Consensus Group on Microembolus Detection was performed using two 2-MHz probes of the pulsed Doppler system MultiDop-T Digital (DWL Compumedics). MRI scans of the brain were carried out using a 1.5-T scanner GE Discovery 450 (GE Healthcare). Detection of carotid stenosis was performed using 3D contrast-enhanced MR angiography. Carotid intima-media thickness (cIMT) was measured with B-mode ultrasound. NC was done using Zeiss device. More than 100 variables were taken into account including cytokines, inflammatory markers, autoantibodies, classical risk factors for atherosclerosis and selected organ manifestations. Statistical analysis was performed with chi2 Yates, chi2 Pearson, rank Spearman correlations tests and logistic regression analysis. Results Factors which significantly correlated with analysed vascular changes including microemboli in TCD, ischaemic changes in MRI and NC abnormalities, were thrombocytopenia (r = 0.47, p = 0.01), C-reactive protein (CRP) (r = 0.51, p = 0.0039) and antiphospholipid antibodies (aPLs) (r = 0.55, p = 0.0015). There was significant association between vascular endothelial growth factor (VEGF) and IL-6 and high cIMT (r = 0.36, p = 0.0492, r = 0.41, p = 0.0239, respectively) as well as NC abnormalities, especially megacapillaries presence (r = 0.38, p = 0.0415, r = 0.42, p = 0.0226, respectively). Additionally, patients with changes in NC significantly more frequently were dyslipidemic (r = 0.56, p = 0.0015), hypertensive (r = 0.41, p = 0.0252) and unveiled high titers of anti-dsDNA (r = 0.37, p = 0.0492) and cardiac involvement (r = 0.38, p = 0.0441). There was also important positive correlation between cIMT and NC abnormalities (r = 0.40, p = 0.0300) as well as microemboli in TCD (r = 0.44, p = 0.0211). Finally, microemboli in TCD were associated with MRI ischaemic changes (r = 0.45, p = 0.0177). Conclusions NC and cIMT provide the optimal protocol to screen SLE patients for cardiovascular risk. CRP, VEGF, IL-6, aPLs and anti-dsDNA seem to be crucial pathogenic factors in micro- and macrovascular impairment development in SLE. Patients with higher cIMT and aPLs should undergo TCD for cerebrovascular risk assessment.

A6.25 Serum concentrations of vascular endothelial growth factor in systemic lupus erythematosus – association with autoantibody profile and cardiovascular involvement

Background and objectives Angiogenesis plays a significant role in the pathogenesis of systemic lupus erythematosus (SLE). Vascular endothelial growth factor (VEGF) is a potent stimulator of angiogenesis as well as vasculogenesis. The study was designed to evaluate the association between VEGF concentrations and immunological parameters, inflammatory markers, classical atherosclerosis risk factors and vascular disorders in SLE patients. Materials and methods The study was performed in 83 patients with SLE and 20 age and gender matched controls. The concentrations of VEGF was determined with ELISA method using R&D Systems tests. The presence of inflammatory markers (ESR, CRP and fibrinogen) and selected autoantibodies - anti-endothelial (AECA), anti-nuclear, anti-phospholipid (aPL) and anti-neutrophil cytoplasmic was evaluated. Classical risk factors for atherosclerosis as well as selected organ manifestations (cardiovascular and central nervous system, lupus nephritis, thromboembolic disorders and vasculitis) were taken into account. Carotid intima-media thickness and atherosclerotic plaques were measured with B-mode ultrasound method. Statistical analysis was performed with chi2Yates, chi2Pearson, rank Spearman correlations tests, logistic regression analysis and multivariate stepwise analysis. Results VEGF levels did not differ significantly between SLE patients and the controls (p > 0.1). The cut-off value of VEGF concentrations was established at 382.4 pg/ml (75- percentile). VEGF levels > 382.4 pg/ml were significantly associated with the elongation of activated partial thromboplastin time (OR = 22.8; 95% CI: 2.3–230.6) and the presence of aPL: anti-prothrombin (aPT) IgA class (OR = 10.7; 95% CI: 2.1–53.4), anti- β2-GPI IgA class (OR = 3.5; 95% CI: 1.1–10.8) and anti-oxidised low density lipoprotein antibodies (OR = 4.8; 95% CI: 1.0–22.8). Myocardial relaxation disorders were significantly more frequent in patients with high concentration of VEGF (OR = 8.0; 95% CI: 1.6–39.5). The low concentration of VEGF significantly decreased the risk of the existence of selected autoantibodies: aPT IgA (OR = 0.18; 95% CI: 0.0–0.72), aβ2-GPI IgA (OR = 0.17; 95% CI: 0.04–0.71), anti-double stranded DNA (OR = 0.31; 95% CI: 0,11–0.91) and AECA (OR = 0.30; 95% CI: 0,11–0.85). Furthermore, they were associated with reduction of the risk of atherosclerotic lesions in iliac arteries (OR = 0.24; 95% CI: 0.0–0.99) and vasculitis development (OR = 0.17; 95% CI = 0.03–0.91). Conclusions 1. High VEGF levels may increase the prothrombotic risk in SLE patients because of the significant association with the presence of antiphospholipid antibodies. 2. The lower concentrations of VEGF significantly decrease the risk of persistence of selected autoantibodies and atherosclerotic lesions as well as vasculitis development in SLE patients.

Recommendations for diagnosis and treatment Fertility, pregnancy and breastfeeding in systemic lupus erythematosus patients

1Katedra i Klinika Reumatologii i Układowych Chorób Tkanki Łącznej, Uniwersytet Medyczny w Lublinie 2Klinika Reumatologii i Chorób Wewnętrznych, Pomorski Uniwersytet Medyczny, Szczecin 3Klinika Chorób Tkanki Łącznej, Instytut Reumatologii, Warszawa 4 Centrum Onkologii – Instytut im. Marii Skłodowskiej-Curie, Warszawa 5Zakład Dydaktyki Ginekologiczno-Położniczej, Warszawski Uniwersytet Medyczny 6 Katedra i Klinika Reumatologii i Chorób Wewnętrznych, Uniwersytet Medyczny, Wrocław 1Chair and Department of Rheumatology and Systemic Connective Tissue Diseases, Medical University of Lublin 2Department of Rheumatology and Internal Diseases, Pomeranian Medical University 3Department of Connective Tissue Diseases, Institute of Rheumatology in Warsaw 4Centre of Oncology, Maria Skłodowska-Curie Institute in Warsaw 5Gynaecology and Obstetrics Didactic Division, Warsaw Medical University 6Chair and Department of Rheumatology and Internal Diseases, Medical University of Wrocław