Systemic Scleroderma Patients Research Articles

Autoantibodies in Serum of Systemic Scleroderma Patients: Peptide-Based Epitope Mapping Indicates Increased Binding to Cytoplasmic Domains of CXCR3.

Systemic sclerosis (SSc) is a severe chronic autoimmune disease with high morbidity and mortality. Sera of patients with SSc contain a large variety of autoantibody (aab) reactivities. Among these are functionally active aab that bind to G protein-coupled receptors (GPCR) such as C-X-C motif chemokine receptor 3 (CXCR3) and 4 (CXCR4). Aab binding to the N-terminal portion of these two GPCRs have been shown to be associated with slower disease progression in SSc, especially deterioration of lung function. Aabs binding to GPCRs exhibit functional activities by stimulating or inhibiting GPCR signaling. The specific functional activity of aabs crucially depends on the epitopes they bind to. To identify the location of important epitopes on CXCR3 recognized by aabs from SSc patients, we applied an array of 36 overlapping 18-20mer peptides covering the entire CXCR3 sequence, comparing epitope specificity of SSc patient sera (N = 32, with positive reactivity with CXCR3) to healthy controls (N = 30). Binding of SSc patient and control sera to these peptides was determined by ELISA. Using a Bayesian model approach, we found increased binding of SSc patient sera to peptides corresponding to intracellular epitopes within CXCR3, while the binding signal to extracellular portions of CXCR3 was found to be reduced. Experimentally determined epitopes showed a good correspondence to those predicted by the ABCpred tool. To verify these results and to translate them into a novel diagnostic ELISA, we combined the peptides that represent SSc-associated epitopes into a single ELISA and evaluated its potential to discriminate SSc patients (N = 31) from normal healthy controls (N = 47). This ELISA had a sensitivity of 0.61 and a specificity of 0.85. Our data reveals that SSc sera preferentially bind intracellular epitopes of CXCR3, while an extracellular epitope in the N-terminal domain that appears to be target of aabs in healthy individuals is not bound by SSc sera. Based upon our results, we could devise a novel ELISA concept that may be helpful for monitoring of SSc patients.

Chronic stress and mental disorders in patients with systemic scleroderma: Results of an interdisciplinary study

To analyze of the prevalence of stressful factors and mental disorders (MDs), as well as their clinical psychopathological and clinical psychological characteristics to improve the comprehensive diagnosis and treatment of systemic scleroderma (SSD). Examinations were performed in 110 patients (predominantly women (n=97 (88.2%); mean age, 49.9±2.47 years) with a documented diagnosis of SSD (its mean duration, 7.25±0.42 years). 62 (56.4%) patients had limited SSD, 36 (32.7%) had diffuse SSD, and 12 (10.9%) had overlap syndrome. The disease was rapidly and slowly progressive in 33 (30%) and 77 (70%) patients, respectively. Oral glucocorticosteroids were used in 99 (90%) patients included in the study, cytotoxic drugs in 66 (60%), plaquenil in 33 (30%); 8 (7%) patients were treated with the biological agent rituximab. All the patients were examined by a psychologist and a psychiatrist. The psychopathological diagnosis of MD was made during a semistructured interview in accordance with the ICD-10 criteria. The Montgomery-Asberg depression and Hamilton anxiety rating scales were used to evaluate the severity of depression and anxiety, respectively. All patients underwent a clinical and psychological examination, including tests assessing memory, attention, and logical thinking, as well as projective techniques. MDs were detected in 91 (83%) patients with SSD. There was a preponderance of depressive disorders in 74 (67.3%) patients: chronic (dysthymia in 33 (30%) patients)) and recurrent (recurrent depressive disorder in 34 (31%)) depressions. Cognitive impairment (CI) of varying severities was diagnosed in 100% of the patients. Schizotypal personality disorder was stated in 44 (40%) patients. 90% of patients were found to have chronic psychic traumas mainly as parental deprivation in childhood (in children less than 11 years of age). 76.7% of the SSD cases developed recurrent episodes of depression in the presence of long-term MD or had a history of the episodes. There was no relationship of MD to gender, age, duration of SSD and its individual clinical manifestations. The nature of SSD treatment did not affect the frequency and spectrum of MD. MDs, predominantly chronic and recurrent depression, and CI are characteristic of most SSD patients. Multiple chronic stressful factors, both previous SSD and those over time, have commonly an impact on the mental health of patients with SSD.

Increased expression of latent TGF-β-binding protein 4 affects the fibrotic process in scleroderma by TGF-β/SMAD signaling

Scleroderma is a fibrosis-related disorder characterized by cutaneous and internal organ fibrosis, and excessive collagen deposition in extracellular matrix (ECM) is a major cause of fibrosis. Transforming growth factor-β (TGF-β)/SMAD signaling has a central role in the pathogenesis of fibrosis by inducing abnormal collagen accumulation in ECM, and latent TGF-β-binding protein 4 (LTBP-4) affects the secretion of latent TGF-β to ECM. A previous study indicated that bleomycin (BLM) treatment increased LTBP-4 expression in lung fibroblasts of Thy-1 knockout mice with lung fibrosis, and LTBP-4 further promoted TGF-β bioavailability as well as SMAD3 phosphorylation. However, the expression and function of LTBP-4 in human scleroderma remain unclear. We aimed to investigate the potential role of LTBP-4 in scleroderma through clinical, in vivo and in vitro studies. LTBP-4 and TGF-β expressions were significantly upregulated in systemic scleroderma (SSc) patients' plasma compared with normal controls (LTBP-4, 1,215±100.2 vs 542.8±41.7 ng/ml, P<0.0001; TGF-β, 1.5±0.2 vs 0.7±0.1 ng/ml, P=0.0031), while no significant difference was found between localized scleroderma (LSc) and normal controls. The plasma concentrations of LTBP-4 and TGF-β were even higher in SSc patients with lung fibrosis (LTBP-4, 1462± 137.3 vs 892.8±113.4 ng/ml, P=0.0037; TGF-β, 2.0±0.4 vs 0.9±0.2 ng/ml, P=0.0212) and esophagus involvement (1390±134.4 vs 940.7±127.0 ng/ml, P=0.0269; TGF-β, 1.9±0.3 vs 0.9±0.2 ng/ml, P=0.0426). The area under receiver operating characteristics (ROC) curve of LTBP-4 was 0.86. Immunohistochemistry measurement also demonstrated a higher LTBP-4 expression in sclerotic skin tissue of LSc and SSc compared with normal controls. More positive fibroblasts were also found in BLM-induced scleroderma mouse model than the saline-treated group. In in vitro studies, knockdown of LTBP-4 in SSc skin fibroblasts prominently reduced downstream COL1A1, COL1A2, and COL3A1 mRNA level by 84%, 82%, and 43%, respectively, and other fibrosis-related genes' expression were also decreased. Furthermore, extracellular TGF-β level and the SMAD2/3 phosphorylation were inhibited through LTBP-4 knockdown treatment, suggesting that the knockdown of LTBP-4 reduced the collagen expression through TGF-β/SMAD signaling pathway. Taken together, these data suggest that LTBP-4 affects fibrotic process in scleroderma, and the high expression of LTBP-4 in SSc plasma may serve as a clinical biomarker in diagnosing this disease. In addition, this study also lays the theoretical foundation for targeting LTBP-4 as treatment of scleroderma.

Pulmonary hypertension in patients with systemic scleroderma

Aim. To evaluate the frequency of pulmonary hypertension (PH) in patients with systemic scleroderma (SSD) and elucidate clinical features of SSD with and without concomitant PH. Materials and methods. The study included 206 patients (189 women and 17 men) with SSD treated in Rheumatological Department, Saratov Regional Hospital. Clinical features, results of echocardiography and external respiration function (ERF) studies were analyzed in SSD patients with and without concomitant PH. Results. PH was documented in 33% of the patients. Their clinical condition, changes in left and right hear as well as ERF were elucidated. Conclusion. The study confirmed the high informative value of echocardiography used to evaluate ERF in patients with SSD and the possibility to apply this method for screening PH in SSD patients. The risk factors of PH in SSD patients include advanced age, apnoea, disturbed ERF, and sclerodactylia.

Abstract 12548: Expression of Let-7 Family microRNAs in Skin Correlates Negatively With Severity of Pulmonary Hypertension in Patients With Systemic Scleroderma

Introduction: Pulmonary hypertension (PH) is a serious complication in patients with systemic scleroderma (SSc), therefore it is important to identify the factors that could predict the presence and progression of PH. Skin biopsy is performed in patients with SSc to examine the type and severity of the disease. MicroRNAs (miRNAs) are potential biomarkers for various cardiovascular diseases including PH. Hypothesis: In patents with systemic scleroderma (SSc), skin miRNAs expression profiles varies depending on the presence or absence of PH. Methods and Results: We determined the skin miRNA expression profile in 15 female SSc patients with (n=6) and without PH (n=9). Plasma BNP level was significantly higher in the PH group than non-PH group, but left ventricular function and dimension were not different between the two groups. A mixture of equal amounts of miRNAs from PH and non-PH patients were prepared and used for miRNA PCR array analysis. The analysis identified 591 upregulated miRNAs and 57 downregulated miRNAs in the PH group. Of these, only miRNAs with a Ct value of less than 35 were subjected to further analysis. When a 1.5-fold difference was considered meaningful, 32 miRNAs were upregulated and 14 miRNAs were downregulated in the PH group. Interestingly, 5 out of 14 downregulated miRNAs belonged to the let-7 family. The results were validated by quantitative real-time PCR with specific primer for each miRNA, which showed significant downregulation of five let-7 family members (let-7a, -7d, -7e, -7f, -7g) in 6 PH compared with 9 non-PH skin samples. Circulating these miRNA levels were trended to decrease in PH group, but it did not reach statistically significant. The expression of let-7b and -7d correlated significantly and negatively with the estimated pulmonary arterial pressure. The expression of let-7a, -7e, -7g, -7f tended to correlate negatively with the estimated pulmonary arterial pressure, albeit insignificantly. Conclusions: The expression level of let-7 family microRNAs in skin are lower in patients with SSc with PH than those without PH. Skin miRNA is a potentially useful marker for evaluation of PH in patients with SSc. Skin biopsy can be considered in patients with collagen diseases to identify the presence and severity of PH.

Expression of Let-7 Family microRNAs in Skin Correlates Negatively with Severity of Pulmonary Hypertension in Patients with Systemic Scleroderma

Pulmonary hypertension (PH) is a serious complication in patients with systemic scleroderma (SSc), therefore it is important to identify the factors that predict the presence and progression of PH. Skin biopsy is performed in patients with SSc to examine the type and severity of the disease. MicroRNAs (miRNAs) are potential biomarkers for various cardiovascular diseases including PH. We determined the skin miRNA expression profile in 15 SSc patients with and without PH. A mixture of equal amounts of miRNAs from PH and non-PH patients were prepared and used for miRNA PCR array analysis. The analysis identified 591 upregulated miRNAs and 57 downregulated miRNAs in the PH group. When a 1.5-fold difference was considered meaningful, 32 miRNAs were upregulated and 14 miRNAs were downregulated in the PH group. Interestingly, 5 out of 14 downregulated miRNAs belonged to the let-7 family. The results were validated by quantitative real-time PCR, which showed significant downregulation of five let-7 family members (let-7a, -7d, -7e, -7f, -7g). The expression levels of let-7d and 7b correlated negatively with pulmonary arterial pressure measured by echocardiography. The results suggest that skin miRNA is a potentially useful marker for the presence and severity of PH in patients with SSc.

Expression of Let-7 family microRNAs in skin correlates negatively with severity of pulmonary hypertension in patients with systemic scleroderma

BackgroundPulmonary hypertension (PH) is a serious complication in patients with systemic scleroderma (SSc), therefore it is important to identify the factors that could predict the presence and progression of PH. Skin biopsy is performed in patients with SSc to examine the type and severity of the disease. MicroRNAs (miRNAs) are potential biomarkers for various cardiovascular diseases including PH. Methods and resultsWe determined the skin miRNA expression profile in 15 SSc patients with (n=6) and without PH (n=9). A mixture of equal amounts of miRNAs from PH and non-PH patients were prepared and used for miRNA PCR array analysis. The analysis identified 591 upregulated miRNAs and 57 downregulated miRNAs in the PH group. Of these, only miRNAs with a Ct value of less than 35 were subjected to further analysis. When a 1.5-fold difference was considered meaningful, 32 miRNAs were upregulated and 14 miRNAs were downregulated in the PH group. Interestingly, 5 out of 14 downregulated miRNAs belonged to the let-7 family. The results were validated by quantitative real-time PCR with specific primer for each miRNA, which showed significant downregulation of five let-7 family members (let-7a, -7d, -7e, -7f, -7g) in 6 PH compared with 9 non-PH skin samples. The expression levels of let-7d and 7b correlated negatively with pulmonary arterial pressure measured by echocardiography. ConclusionsThe results suggest that skin miRNA is a potentially useful marker for the presence and severity of PH in patients with SSc.

Application of cloud database in the management of clinical data of patients with skin diseases.

To evaluate the needs and applications of using cloud database in the daily practice of dermatology department. The cloud database was established for systemic scleroderma and localized scleroderma. Paper forms were used to record the original data including personal information, pictures, specimens, blood biochemical indicators, skin lesions,and scores of self-rating scales. The results were input into the cloud database. The applications of the cloud database in the dermatology department were summarized and analyzed. The personal and clinical information of 215 systemic scleroderma patients and 522 localized scleroderma patients were included and analyzed using the cloud database. The disease status,quality of life, and prognosis were obtained by statistical calculations. The cloud database can efficiently and rapidly store and manage the data of patients with skin diseases. As a simple, prompt, safe, and convenient tool, it can be used in patients information management, clinical decision-making, and scientific research.

Anti-PM/Scl antibodies are found in Japanese patients with various systemic autoimmune conditions besides myositis and scleroderma.

IntroductionAnti-PM/Scl antibodies are associated with polymyositis (PM)/systemic scleroderma (SSc) overlap syndromes and are also found in other systemic autoimmune diseases. Although anti-PM/Scl reactivity is found in 3-11% of PM or SSc patients and in approximately 25% of PM/SSc overlap patients, previous large studies of Japanese patients with scleroderma reported that anti-PM/Scl are not found in Japanese patients at all. The PM/Scl autoantigen complex comprises 11–16 different polypeptides; ELISA with PM1-α peptide, which is a major epitope of the PM/Scl complex, has frequently been used for the detection of these antibodies in recent studies. However, no ELISA kit is commercially available in Japan.MethodsIn this study, we developed an immunoassay for measuring antibodies against recombinant PM/Scl-100 and PM/Scl-75 polypeptides, which are the two major targets of the complex, and we investigated their presence in 600 Japanese patients with various systemic autoimmune conditions. Immunoprecipitation analysis using the recombinants in addition to traditional radiolabeled cell extracts were also applied to ELISA-positive sera.ResultsIn ELISA, 11 patients were positive for anti-PM/Scl-100 antibodies and 7 of these 11 patients were also positive for anti-PM/Scl-75 antibodies. Immunoprecipitation analysis using the recombinants in addition to traditional radiolabeled cell extracts confirmed that 9 out of these 11 patients immunoprecipitated the typical sets of PM/Scl proteins. In total, 4/16 (25%) undifferentiated connective tissue disease (UCTD) patients, 3/126 (2.4%) dermatomyositis patients, 1/223 (0.4%) SSc patients, 1/88 (1.1%) Sjögren’s syndrome patients, 0/123 patients with systemic lupus erythematosus, 0/17 patients with overlap syndrome and 0/7 patients with PM were judged to be positive for anti-PM/Scl antibodies.ConclusionsThis is the first report of Japanese autoimmune patients with anti-PM/Scl antibodies. In Japanese patients, anti-PM/Scl antibodies are only very rarely found, and they are not always specific for dermatomyositis (DM) or SSc; they are also present in various autoimmune conditions with the highest prevalence being in UCTD. All anti-PM/Scl-positive DM cases are complicated with interstitial lung disease and/or cancer, while no life-threatening involvement was found in other anti-PM/Scl-positive cases. Further studies on larger cohorts are necessary to define the clinical significance of anti-PM/Scl antibodies in autoimmune diseases.

Abstract 11127: Let-7 Family MicroRNAs Expression in Subcutaneous Skin Negatively Correlate With Severity of Pulmonary Hypertension in Patients With Systemic Scleroderma

Introduction: MicroRNAs (miRNAs) have been shown to play an important role in various cardiovascular diseases. However, its roles in the pathogenesis of connective tissue disease-associated pulmonary hypertension (PH) are not clear. Hypothesis: In patents with systemic scleroderma (SSc), subcutaneous miRNAs expression profiles are different depending on the presence or absence of PH. Methods and Results: Data of 16 SSc patients who underwent subcutaneous tissue biopsy were available for retrospective analysis. To determine the changes in subcutaneous miRNAs expression profiles in SSc-associated PH, a mixture of equal amount of subcutaneous miRNAs from 6 PH or 6 non-PH were prepared. We performed miRNA PCR array analysis, and the miRNA expression was comprehensively compared between PH and non-PH group. We found that 591 miRNAs were upregulated, and 57 miRNAs were downregulated in PH group (Figure). Among them, only miRNAs which showed a Ct value less than 35 were considered for further analysis. When a 1.5-fold difference was considered meaningful, 32 miRNAs were upregulated, and 14 miRNAs were down regulated in PH group. Interestingly, 5 out of 14 downregulated miRNAs belong to let-7 family (let-7a, -7b, -7d, -7f, -7g). We validated the results by quantitative real-time PCR with specific primer for each miRNAs individually and found that 4 let-7 family members (let-7a, -7d, -7f, -7g) were significantly downregulated in 6 PH compared with those in 10 non-PH skin. The expression level of let-7d was negatively correlated with pulmonary arterial pressure measured by echocardiography. Conclusions: The expression level of let-7 family microRNAs in subcutaneous skin are lower in patients with SSc with PH than those without PH. Let-7 family microRNAs expression in subcutaneous skin negatively correlate with severity of PH in patients with SSc. Subcutaneous miRNA could be a useful marker to identify the presence and severity of PH in patients with SSc.

Let-7 Family microRNAs Expression in Subcutaneous Skin Negatively Correlate with Severity of Pulmonary Hypertension in Patients with Systemic Scleroderma

Introduction: MicroRNAs (miRNAs) have been shown to play an important role in various cardiovascular diseases. However, its roles in the pathogenesis of connective tissue disease-associated pulmonary hypertension (PH) are not clear. Hypothesis: In patents with systemic scleroderma (SSc), subcutaneous miRNAs expression profiles are different depending on the presence or absence of PH. Methods and Results: Data of 16 SSc patients who underwent subcutaneous tissue biopsy were available for retrospective analysis. To determine the changes in subcutaneous miRNAs expression profiles in SSc-associated PH, a mixture of equal amount of subcutaneous miRNAs from 6 PH or 6 non-PH were prepared. We performed miRNA PCR array analysis, and the miRNA expression was comprehensively compared between PH and non-PH group. We found that 591 miRNAs were upregulated, and 57 miRNAs were downregulated in PH group (Figure). Among them, only miRNAs which showed a Ct value less than 35 were considered for further analysis. When a 1.5-fold difference was considered meaningful, 32 miRNAs were upregulated, and 14 miRNAs were down regulated in PH group. Interestingly, 5 out of 14 downregulated miRNAs belong to let-7 family (let-7a, -7b, -7d, -7f, -7g). We validated the results by quantitative real-time PCR with specific primer for each miRNAs individually and found that 4 let-7 family members (let-7a, -7d, -7f, -7g) were significantly downregulated in 6 PH compared with those in 10 non-PH skin. The expression level of let-7d was negatively correlated with pulmonary arterial pressure measured by echocardiography. Conclusions: The expression level of let-7 family microRNAs in subcutaneous skin are lower in patients with SSc with PH than those without PH. Let-7 family microRNAs expression in subcutaneous skin negatively correlate with severity of PH in patients with SSc. Subcutaneous miRNA could be a useful marker to identify the presence and severity of PH in patients with SSc. ![][1] [1]: /embed/graphic-1.gif

ANTI- ANNEXIN 1 ANTIBODIES CORRELATION BETWEEN ANTIBODIES LEVEL AND DISEASE ACTIVITY IN SOME AUTOIMMUNE CONNECTIVE TISSUE DISEASES

Annexin 1 is an anti-inflammatory protein, exerts a regulatory effect in both innate and adaptive immune responses. Autoantibodies against annexin 1 have been recognized in some patients with autoimmune connective tissue diseases. Aim: to detect presence of anti-annexin 1 antibodies in patients with rheumatoid arthritis, scleroderma, systemic and discoid lupus erythematosus and to evaluate whether there is a correlation between the antibodies levels and disease activity. Methods: determining the levels of IgM antibodies in serum samples by ELISA and correlating them with the disease activity, which is evaluated using specific activity index for each disease. Results: levels of anti- annexin 1 antibodies were found to be significantly higher in diseased groups when compared to normal healthy controls. When correlating antibodies levels with disease activity, rheumatoid arthritis and discoid lupus patients showed no significant correlation, however systemic lupus and scleroderma patients showed significant correlation between antibodies levels and disease activity. Conclusion: anti- annexin 1 antibodies might be a promising aid in diagnosis of these diseases and could have a prognostic capacity in some autoimmune connective tissue diseases. Key words: Annexin 1- autoantibodies- autoimmune connective tissue diseases.

Epstein–Barr virus antibodies mark systemic lupus erythematosus and scleroderma patients negative for anti‐DNA

Systemic lupus erythematosus (SLE) is an autoimmune disease that can attack many different body organs; the triggering event is unknown. SLE has been associated with more than 100 different autoantibody reactivities - anti-dsDNA is prominent. Nevertheless, autoantibodies to dsDNA occur in only two-thirds of SLE patients. We previously reported the use of an antigen microarray to characterize SLE serology. We now report the results of an expanded study of serology in SLE patients and scleroderma (SSc) patients compared with healthy controls. The analysis validated and extended previous findings: two-thirds of SLE patients reacted to a large spectrum of self-molecules that overlapped with their reactivity to dsDNA; moreover, some SLE patients manifested a deficiency of natural IgM autoantibodies. Most significant was the finding that many SLE patients who were negative for autoantibodies to dsDNA manifested abnormal antibody responses to Epstein-Barr virus (EBV): these subjects made IgG antibodies to EBV antigens to which healthy subjects did not respond or they failed to make antibodies to EBV antigens to which healthy subjects did respond. This observation suggests that SLE may be associated with a defective immune response to EBV. The SSc patients shared many of these serological abnormalities with SLE patients, but differed from them in increased IgG autoantibodies to topoisomerase and centromere B; 84% of SLE patients and 58% of SSc patients could be detected by their abnormal antibodies to EBV. Hence an aberrant immune response to a ubiquitous viral infection such as EBV might set the stage for an autoimmune disease.

Is exercise-induced pulmonary hypertension an early sign of onset of resting pulmonary hypertension in patients with scleroderma?

Background: Resting pulmonary hypertension (PH) is a powerful determinant of poor outcome in patients with systemic scleroderma (SSc). In various cardiac diseases, exercise-induced PH (EIPH) is considered as an early stage of resting PH, and may predict the occurrence of adverse events. We hypothesized that the presence of EIPH is a predictor of the onset of resting PH in patients with SSC. Methods: 33 patients with diagnosis of SSc disease were prospectively submitted to comprehensive both resting and exercise stress echocardiography between January 2009 and November 2012. Resting PH was defined as a transtricuspid pressure gradient (TTG) ≥ 30mmHg, and EIPH as TTG ≥ 40 mmHg Patients were prospectively follow-up and regularly performed resting echocardiography. Results: There was a significant increase in TTG from rest to exercise (21.5±7.7 vs. 37.5±15.2, p<0.001). There were 2 patients with baseline resting PH (6%) and 15 patients with EIPH (45%). Of note, all patients with resting PH have developed EIPH. Mean follow-up was 25±13 months (from 1 to 52) and 6 patients have experienced resting PH during this period, resulting in a mean time interval between baseline echocardiography and occurrence of resting PH of 8±7 months (from 1 to 18). Of interest, all patients who developed resting PH during the follow-up have baseline EIPH (60%). In contrast, none of patients without baseline EIPH developed resting PH during the follow-up (0%). Although patients who developed resting PH had significant higher baseline resting TTG (26.6±4.0 vs. 17.8±5.7, p=0.03), the difference in baseline exercise TTG was markedly higher (53.7±14.0 vs. 31.4±11.9, p=0.001). Using logistic regression, after adjustment for age, baseline EIPH was independently associated with the occurrence of resting PH during the follow-up (p=0.0014). Conclusion: EIPH may be frequent in SSc patients and could be a predictive factor of rapid and early onset of resting PH. Exercise stress echocardiography may be useful to identify subset of patients at high risk to developed resting PH and thus, could benefit from more aggressive therapeutic strategy. Nevertheless, further data in larger cohort are needed to confirm our results.

THU0005 Investigating the Plasma Cell Signature in Autoimmune Disease

BackgroundProduction of pathogenic autoantibodies by self-reactive plasma cells (PC) is a hallmark of autoimmune diseases; thus, PC levels could be associated with efficacy of B-cell depleting therapies. Additionally, investigating the...

Anaemia in a Patient with Diffuse Systemic Scleroderma

We hereby present a case of anaemia in a 73 years old patient with known past medical history of diffuse systemic scleroderma, who presented with acute onset of dizziness and haemetemesis. Blood tests revealed sudden drop of haemoglobin and an urgent gastroscopy revealed gastric antral vascular ectasia (GAVE) or “watermelon stomach”. GAVE is a rare but well recognised cause of acute bleeding in systemic scleroderma patients and should be kept as a differential diagnosis in the work up of anaemia in these patients.

Antifibrotic effects of crocetin in scleroderma fibroblasts and in bleomycin-induced sclerotic mice

OBJECTIVE:To investigate the antifibrotic effects of crocetin in scleroderma fibroblasts and in sclerotic mice.METHODS:Skin fibroblasts that were isolated from three systemic scleroderma (SSc) patients and three healthy subjects were treated with crocetin (0.1, 1 or 10 μM). Cell proliferation was measured with an MTT assay. Alpha-smooth muscle actin was detected via an immunohistochemical method. Alpha 1 (I) procollagen (COL1A1), alpha 1 (III) procollagen (COL3A1), matrix metalloproteinase (MMP)-1 and tissue inhibitor of matrix metalloproteinase (TIMP)-1 mRNA levels were measured using real-time PCR. SSc mice were established by the subcutaneous injection of bleomycin. Crocetin (50 mg/kg/d) was injected intraperitoneally for 14 days. Dermal thickness and lung fibrosis were assessed with Masson's trichrome staining. Plasma ET-1 was detected with an enzyme-linked immunosorbent assay (ELISA). Skin and lung ET-1 and COL1A1 mRNA levels were measured via real-time PCR.RESULTS:Crocetin inhibited the proliferation of SSc and normal fibroblasts, an effect that increased with crocetin concentration and incubation time. Crocetin decreased the expression of α-SMA and the levels of mRNA for COL1A1, COL3A1 and matrix metalloproteinase-1, while crocetin increased TIMP-1 mRNA levels in both SSc and normal fibroblasts. Skin and lung fibrosis was induced, and the levels of ET-1 in the plasma, skin and lungs were elevated in bleomycin-injected mice. Crocetin alleviated the thickening of the dermis and lung fibrosis; decreased COL1A1 mRNA levels in the skin and lung; and simultaneously decreased ET-1 concentrations in the plasma and ET-1 mRNA levels in the skin and lungs of the bleomycin-induced sclerotic mice, especially during the early phase (weeks 1-3).CONCLUSION:Crocetin inhibits cell proliferation, differentiation and collagen production in SSc fibroblasts. Crocetin alleviates skin and lung fibrosis in a bleomycin-induced SSc mouse model, in part due to a reduction in ET-1.

THE REAL PREVALENCE OF EROSIVE ESOPHAGITIS AND BARRETT'S ESOPHAGUS IN SYSTEMIC SCLERODERMA: DATA FROM 12-MONTHS PROSPECTIVE STUDY

Complicated forms of reflux-esophagitis, i.e., erosive esophagitis and Barrett's esophagus (BE) — are common types of visceral pathology in systemic scleroderma (SSD), which require adequate therapy and follow up. Although real prevalence of esophageal involvement in SSD in Russian patients remains uncertain. Objective — to identify prevalence of erosive esophagitis and BE, and to quantify gastro-intestinal (GI) symptoms in patients with SSD. Material and methods. During 1 year (December 2009 — January 2011) all consecutive SSD patients, hospitalized to FSBI «SRIR» RAMS, after signing informed consent, were subjected to esophagogastroduodenoscopy with biopsy of esophageal mucosa in upper 1/3. Totally 123 patients were examined (96,8% females, 3,2% males, aged 50,5±13,1 years). Esophageal mucous was evaluated for presence of pathologic changes and BE (intestinal metaplasia in biopsy samples was a BE diagnostic criterion). SODA questionnaire was used to quantify GIT symptoms Results. Erosive esophagitis was detected in 30 (24,3%) patients, BE — in 11 (8,9%). In 80% of patients marked changes in esophageal mucosa were associated with typical symptoms (heartburn, regurgitaion, dysphagia), while in some cases (in 3 patients) erosive esophagitis and BE were asymptomatic. Quantitative evaluation of symptoms with SODA questionnaire demonstrated clear correlation between subjective assessment and severity of esophageal pathologic changes. In patients with erosive gastritis and BE the SODA «pain» and «non-pain» parameters scores were significantly higher and satisfaction in dyspepsia management was lower (p<0,05), then in individuals without erosions and mucosal inflammation. Here was no clear correlation between esophageal pathology and SSD type (limited, diffuse), age, duration of the disease, presence of pulmonary interstitial lesion and Sjogren's syndrome. Patients with erosive esophagitis were significantly more often (36,6%) using proton pomp inhibitors as compared to individuals without apparent esophageal pathology (22,0%; p<0,001). Conclusion. Erosive esophagitis is diagnosed in each 4th patient with SSD.EB is also quite common in this pathology, predetermining the necessity of regular endoscopic check-ups in all SSD patients. SODA questionnaire is a useful tool to evaluate the severity of upper GIT symptoms in SSD patients during follow up.

Drug compliance in patients with systemic scleroderma

Although drug compliance is a crucial component of treatment effectiveness in chronic diseases, it has never been evaluated in patients with systemic scleroderma. Therefore, the aim of this descriptive study was to determine the drug compliance rate in systemic scleroderma patients and to identify risk factors for noncompliance in these patients. A cross-sectional observational study was conducted. All patients with systemic scleroderma (n = 41) who visited a rheumatic center and signed an informed consent form were included. Data were obtained during structured interviews with patients and from medical records. The Compliance Questionnaire Rheumatology (CQR) was used to determine patient compliance. The relationships between compliance rate and demographic and clinical characteristics were examined. The mean CQR score was 75 %. Based on a dichotomous rating, only 42 % of the patients achieved a satisfactory compliance rate (≥80 %). No relationships between various demographic and clinical characteristics and CQR score expressed as continuous or dichotomous variables were found. This study represents the first evaluation of drug compliance in patients with systemic scleroderma. Many noncompliant patients were identified, but no common risk factors for noncompliance were discovered. The reasons for noncompliance seem to depend on the personal features of the patients.

Prévalence et valeur diagnostique des anticorps antinucléaires de spécificité antigénique indéterminée : étude rétrospective à propos d’une série de 90 patients

PurposeThe objective of this study was to determine the clinical relevance and the diagnostic significance of positive antinuclear antibodies (ANA) without identified antigenic target by the usual characterization technique. Patients and methodsRetrospective study conducted in the Laboratory of Immunology of Habib Bourguiba Hospital (Sfax, Tunisia) during 18months. The inclusion criteria were the presence of an ANA titer greater or equal to 1/320 with negative characterization result. ANA screening was performed by indirect immunofluorescence (IIF) on Hep2 cells. Each positive serum was tested by IIF on Crithidia luciliae (anti-native DNA) and by immunodot (anti-nucleosome, anti-histone, anti-Sm, anti-RNP, anti-SSA, anti-SSB, anti-Scl 70, anti-PM-Scl, anti-Jo1, anti-PCNA and anti-ribosomal protein). Sera of systemic lupus erythematosus (SLE), myositis, and scleroderma patients were tested for anti-Ku, anti-PL7, anti-PL12 and anti-Ro-52 using dot myositis. ResultsSera of 90 patients were studied: 18 men and 72 women (average age: 44years). Drug-induced ANA was found in eight patients. The most frequent clinical symptoms were joint (56.7%), cutaneous (54.4%) and constitutional symptoms (45.6%). The diagnosis of an autoimmune disease was suspected in 49 patients (54.5%) and confirmed in 30 (33.3%) including 20 cases of connective tissue disease: myositis (n=6), scleroderma (n=5), Sjögren's syndrome (n=3), SLE (n=4), rheumatoid arthritis (n=6) and antiphospholipid syndrome (n=4). Other autoimmune diseases were less frequent. The anti-Ku antibody was detected in the majority of patients with connective tissue disease. The diagnosis of non-autoimmune diseases was established in 25.5% of patients. Eighteen patients (20%) had no diagnosis orientation. ConclusionOur study demonstrated the diagnostic value of the presence of ANA even in the absence of known antigenic target, confirmed the role of the IIF as “gold standard” test for ANA screening, and suggested the usefulness of the addition of Ku antigen in the immunodot classic profile.