Anti-nucleosome Antibodies Research Articles

Antinucleosome antibodies in systemic lupus erythematosus patients: Relation to disease activity and lupus nephritis

Aim of the workTo evaluate the level of anti-nucleosome (anti-NCS) antibodies in systemic lupus erythematosus (SLE) patients and study their association with disease activity and lupus nephritis. Patients and methodsThe study was carried out on 50 SLE patients; 47 females and 3 males and 30 matched controls. The SLE disease activity index (SLEDAI) was assessed. Plasma levels of anti-NCS antibodies were measured. Renal biopsy was done in those with lupus nephritis. ResultsThe mean age of patients was 28.3 ± 12.9 years; disease duration was 5.5 ± 6.02 years and the SLEDAI.5 ± 7.8. LN was present in 60% of the patients. Anti-nuclear antibodies (ANA) were positive in 48 (96%) and the anti-double stranded deoxyribonucleic acid positive in 38 (76% of the patients. The anti-NCS antibodies level was significantly increased in the patients (470.1 ± 369.4 U/ml) compared to the control (36.43 ± 14.58 U/ml) (p = 0.0001). The anti-NCS was detected in 48/50(96%) patients at a cut-off of 65 U/ml. There was no significant difference in the anti-NCS antibodies levels between those with nephritis (456.53 ± 362.59 U/ml) and those without (490.4 ± 387.96 U/ml) (p = 0.84). A significant correlation was present between the anti-NCS antibodies and photosensitivity (p = 0.032), ESR (p = 0.03), complement (C3) consumption (p = 0.01) and urinary casts (p = 0.04). A non-significant correlation was detected between level of anti-NCS and grades of nephritis (p = 0.49) or SLEDAI (p = 0.09). ConclusionAnti-NSC antibodies could be a useful addition to the laboratory tests that can help in the diagnosis of SLE. Also, it has a significant association to photosensitivity but no relation to with disease activity or renal involvement.

Antinucleosome Antibody in Children and Adolescents with Systemic Lupus Erythematosus

AbstractBackground: SLE is an autoimmune disease of unknown etiology characterized by the production of abroad and heter-ogeneous group of autoantibodies. These autoantibodies are directed to nuclear, cytoplasmic, and cellular membrane antigens. Recently, it was proposed that the nucleosome is the principal antigen in the pathophysiology of SLE, and that anti-Nuc antibodies are associated with organic damage.Aim of the Study: The aim of this work was to study the potential utility of serum levels of anti- nucleosome antibodies as a diagnostic tool and a disease activity marker in children and adolescents with systemic lupus erythematosus.Patients and Methods: The study was carried out on forty five patients with SLE who attended to the outpatient clinic and inpatient of Pediatric Nephrology matched age and sex served as a control group. All studied children were subjected to full history, complete physical examination, SLEDAI score, routine laboratory investigations and anti-dsDNA and anti-nucleosome antibody IgG assay. Data was analyzed by using SPSS.Results: The mean serum level of anti-Nuc antibody was significantly higher in patients than controls (p-value <0.001). But there was no significant difference between patients' subgroups. There was a weak correlation between serum anti-Nuc antibody and SLEDAI score (r: 0.213) but strong corre-lation between anti-dsDNA antibody and SLEDAI score (r: 0.711). Anti-Nuc antibody showed higher sensitivity but equal specificity to anti-dsDNA antibody for the diagnosis of SLE.Conclusion: Anti-nucleosome antibodies are superior to anti-dsDNA antibodies in the diagnosis of SLE especially in anti-dsDNA negative patients as they have higher sensitivity but as regard to disease activity antidsDNA antibody is more accurate.

Clinical and immunological characteristics of Polish patients with systemic lupus erythematosus.

Systemic lupus erythematosus (SLE) is a chronic autoimmune disease with varied clinical manifestations, which creates difficulties and delays in establishing a diagnosis. The aim of this study was to evaluate the prevalence and nature of the clinical symptoms of SLE, both at the onset of the disease and in its further course. An attempt to assess the immunological characteristics of the patients and to analyze autoantibodies variability over time was also made. This retrospective study included 71 Caucasian patients, 63 women and 8 men, meeting the criteria for diagnosis of SLE according to ACR. The ratio of women to men was approximately 7.9:1. The average age of the onset of SLE was 31.5 years. The average time from the onset of symptoms to diagnosis was 5 years. The most common first manifestation of SLE were joint and muscles symptoms - 71.8%, skin lesions - 69.0%, fever - 57.7%. The main symptoms in the further course of the disease were neurological disorders - 69.0%, joint and muscle changes - 67.7%, and general symptoms - 59.2%. There was an increase in the incidence of renal involvement and neurological symptoms throughout the disease course. The most commonly detected antibodies were anti-dsDNA - 47.9%, anti-Ro/SSA - 40.8%, anti-nucleosomal antibodies - 29.6%, and lupus anticoagulant - 22.5%. A panel of antibodies typically did not change. There is no typical clinical picture of SLE, the population suffering from this disease is very various. Therefore, early and accurate diagnosis can be a big challenge for any clinician, which justifies the need for this type of study to better characterize the disease.

Defining the epichromatin epitope

ABSTRACTEpichromatin is identified by immunostaining fixed and permeabilized cells with particular bivalent anti-nucleosome antibodies (mAbs PL2-6 and 1H6). During interphase, epichromatin resides adjacent to the inner nuclear membrane; during mitosis, at the outer surface of mitotic chromosomes. By STED (stimulated emission depletion) microscopy, PL2-6 stained interphase epichromatin is ∼76 nm thick and quite uniform; mitotic epichromatin is more variable in thickness, exhibiting a “wrinkled” surface with an average thickness of ∼78 nm. Co-immunostaining with anti-Ki-67 demonstrates Ki-67 deposition between the PL2-6 “ridges” of mitotic epichromatin. Monovalent papain-derived Fab fragments of PL2-6 yield a strikingly different punctate “chromomeric” immunostaining pattern throughout interphase nuclei and along mitotic chromosome arms. Evidence from electrophoretic mobility shift assay (EMSA) and from analytical ultracentrifugation characterize the Fab/mononucleosome complex, supporting the concept that there are two binding sites per nucleosome. The peptide sequence of the Hv3 region (heavy chain variable region 3) of the PL2-6 antibody binding site strongly resembles other nucleosome acidic patch binding proteins (especially, LANA and CENPC), supporting that the nucleosome acidic patch is included within the epichromatin epitope. It is speculated that the interphase epichromatin epitope is “exposed” with favorable geometric arrangements for binding bivalent PL2-6 at the surface chromatin; whereas, the epitope is “hidden” within internal chromatin. Furthermore, it is suggested that the “exposed” nucleosome surface of mitotic epichromatin may play a role in post-mitotic nuclear envelope reformation.

Cardiac tamponade in systemic lupus erythematosus.

Cardiac tamponade is a rare but life-threatening complication of systemic lupus erythematosus (SLE). To describe incidence, risk factors and treatment of cardiac tamponade in a large cohort of Indian patients with SLE. This retrospective study was conducted at the Department of Rheumatology, IPGMER, Kolkata, India from May 2014 to December 2016 on admitted patients with SLE. Lupus-related serositis was diagnosed after excluding other causes, such as infection, malignancy or heart failure. Of 409 patients with SLE, pericarditis was diagnosed in 25.4% (104/409) and cardiac tamponade in 5.9% (24/409). Tamponade was the presenting feature of SLE in 50% (12/24). Tamponade occurred in 77.8% (14/18) of large effusions and in 11.63% (10/86) of small-to-moderate effusions. The commonest autoantibody in serum and pericardial fluid was anti-nucleosme antibody. Large pericardial effusion (>20 mm) (Odd's ratio (OR): 93.2, 95% confidence interval (CI): 11.1-782.5, P < 0.001) predicted tamponade. In the subset of patients with small-to-moderate sized pericardial effusion, tamponade was associated with pleuritis (OR: 44.5, 95% CI: 1.6-1243, P = 0.025), anti-nucleosome antibody (OR: 42.9, 95% CI: 1.6-1176, P = 0.026) and size of pericardial effusion (OR: 1.36, 95% CI: 1.04-1.76, P = 0.025). Repeated pericardiocentesis was required in 3 patients and one needed surgical intervention. Immunosuppressives used were: prednisolone with monthly intravenous cyclophosphamide (in 33.33%) and intravenous methylprednisolone with monthly cyclophosphamide (in 50%). Pleuritis, anti-nucleosome antibody and size of pericardial effusion predicted development of tamponade. High dose immunosuppression (methylprednisolone and IV cyclophosphamide) alleviated need for surgery in majority.

Serum levels of adiponectin and leptin as biomarkers of proteinuria in lupus nephritis.

IntroductionThere are controversial results about the role of serum leptin and adiponectin levels as biomarkers of the severity of proteinuria in lupus nephritis.ObjectiveThe aim of this study was to evaluate the relationship between serum leptin and adiponectin levels with severity of proteinuria secondary to lupus nephritis (LN).MethodsIn a cross-sectional study, 103 women with systemic lupus erythematosus (SLE) were evaluated for kidney involvement. We compared 30 SLE patients with LN, all of them with proteinuria, versus 73 SLE patients without renal involvement (no LN). A comprehensive set of clinical and laboratory variables was assessed, including serum levels of leptin and adiponectin by ELISA. Multivariate analyses were used to adjust for potential confounders associated with proteinuria in LN.ResultsWe found higher adiponectin levels in the LN group compared with the no LN group (20.4 ± 10.3 vs 15.6 ± 7.8 μg/mL; p = 0.02), whereas no differences were observed in leptin levels (33.3 ± 31.4 vs 22.5 ± 25.5 ng/mL; p = 0.07). Severity of proteinuria correlated with an increase in adiponectin levels (r = 0.31; p = 0.001), but no correlation was observed with leptin. Adiponectin levels were not related to anti-dsDNA or anti-nucleosome antibodies. In the logistic regression, adiponectin levels were associated with a high risk of proteinuria in SLE (OR = 1.06; 95% CI 1.01–1.12; p = 0.02). Instead, leptin was not associated with LN.ConclusionThese findings indicate that adiponectin levels are useful markers associated with proteinuria in LN. Further longitudinal studies are required to identify if these levels are predictive of renal relapse.

IFN-\u03bb1 with Th17 axis cytokines and IFN-\u03b1 define different subsets in systemic lupus erythematosus (SLE)

BackgroundInterferon (IFN)-α is thought to have a pivotal role in systemic lupus erythematosus (SLE), and type III IFNs (IFN-λ) were recently also associated with SLE. In this study, we measured levels of IFN-α, IFN-λ1, and related cytokines, such as IL-17A, IL-23, and interferon-γ-induced protein 10 (IP-10), in a Karolinska University Hospital cohort of patients with SLE and control subjects. The objective of the study was to investigate if cytokine measurements could identify different subsets of patients with active SLE and higher disease damage.MethodsWe included 261 patients with SLE and 261 population control subjects. All participants underwent a standardized clinical examination. Medical files were reviewed. Patients with SLE were assessed for current organ manifestations, disease activity, and damage. Routine blood parameters, complement levels, and serology were analyzed at the time of inclusion. Levels of IFN-λ1, IFN-α, IL-17A, IL-23, and IP-10 were measured by enzyme-linked immunosorbent assay.ResultsIFN-λ1 and IFN-α were detected in 29% and 44% of patients, respectively, but their levels did not correlate. High serum levels of IFN-λ1 were positively associated with antinucleosome antibodies and lymphopenia but negatively with musculoskeletal damage. Positive correlations between levels of IFN-λ1, IL-17A, and IL-23 were observed. Patients with high levels of these three cytokines had more disease damage, especially renal impairment. High levels of IFN-α were associated with mucocutaneous disease; leukopenia; and low complement, Ro/SSA, and La/SSB. Vascular events and antiphospholipid antibodies were uncommon. We identified two subgroups with high disease activity: one with double-high IFN-λ1 and IFN-α and another with IP-10high. The former had more neuropsychiatric manifestations, and the latter had more arthritis. Increased levels of both types I and III IFNs were found in a proportion of population control subjects. Therefore, high IFN levels do not seem to be SLE-specific biomarkers.ConclusionsMeasurements of circulating IFN-λ1 and IFN-α define subsets of patients with SLE with different characteristics. Levels of IFN-λ1 correlate with T-helper type 17 cytokines and identify a subgroup with more damage. High disease activity is associated with either simultaneous upregulation of IFN-λ1 and IFN-α or independently with IP-10. Our findings could be of major importance when tailoring therapy for patients with SLE with agents targeting IFN pathways.

Anti-nucleosome antibodies in systemic lupus erythematosus patients: Relation to anti-double stranded deoxyribonucleic acid and disease activity

Aim of the workTo measure the level of anti-nucleosome (anti-NCS) antibodies in systemic lupus erythematosus (SLE) patients and to evaluate their relation with anti-double stranded deoxyribonucleic acid (anti-dsDNA) antibodies and SLE disease activity. Patients and methods66 Egyptian SLE patients were investigated for the detection of anti-NCS antibodies and anti-dsDNA antibodies. Disease activity was assessed using the SLE disease activity index (SLEDAI) and the European consensus lupus activity measurement (ECLAM). ResultsThe median age of the patients was 25.5years (12–48years) and disease duration 3years (1month to 26years). anti-NCS antibody was found in 48 (72.7%) patients. Non-significant difference was found between both those positive or negative anti-NCS antibodies regarding the clinical features apart from fever (p=0.019). Lupus nephritis was present in 35/48(72.9%) of those with positive and in 11/18 (61.1%) of those with negative anti-NCS (p=0.35) A significant correlation was found between anti-NCS antibodies with SLEDAI (r=0.36, p=0.003) and ECLAM (r=0.29, p=0.019). No significant relation was found between anti-NCS antibodies and clinical features of SLE, apart from fatigue (r=0.3, p=0.015). A significant correlation with hypocomplementemia (C3 r=−0.37, p=0.002 and C4 r=−0.32, p=0.018) and anaemia (r=−0.32, p=0.009). anti-dsDNA antibodies were detected in 35(53%) SLE patients; 70.8% of those with positive and 5.6% of those with negative anti-NCS antibodies. ConclusionAnti-NCS antibodies could play a role in the pathogenesis of SLE and is related to disease activity. Its association with anti-dsDNA antibodies and its presence in those with negative anti-ds DNA may aid in the diagnosis of SLE.

Serum levels of P-glycoprotein and persistence of disease activity despite treatment in patients with systemic lupus erythematosus.

Around 25% of patients with systemic lupus erythematosus (SLE) could be refractory to conventional therapies. P-glycoprotein expression on cell surface has been implied on drug resistance, however, to date, it is unknown if P-gp serum levels are associated with SLE disease activity. Evaluate the association of serum P-gp levels and SLE with disease activity despite treatment. A cross-sectional study was conducted on 93 female SLE patients, all receiving glucocorticoids at stable doses for the previous 6months before to baseline. SLE patients were classified into two groups: (a) patients with active disease [SLE disease activity index (SLEDAI)≥3] despite treatment, and (b) patients with inactive disease (SLEDAI<3) after treatment. Forty-three healthy females comprised the control group. Serum P-gp, anti-DNA, and both anti-nucleosome antibody levels were measured using ELISA. Active-SLE patients despite treatment had higher P-gp levels compared with inactive-SLE after treatment (78.02ng/mL±114.11 vs. 33.75ng/mL±41.11; p=0.018) or versus reference group subjects (30.56ng/mL±28.92; p=0.011). P-gp levels correlated with the scores of SLEDAI (r=0.26; p=0.01), Mexican-SLEDAI (MEX-SLEDAI) (r=0.32; p=0.002), SLICC/ACR damage index (r=0.47; p<0.001), and with prednisone doses (r=0.33; p=0.001). In the multivariate model, the high P-gp levels were associated with SLICC/ACR score (p=0.001), and SLEDAI score (p=0.014). Our findings support a relationship between serum P-gp levels and SLE with disease activity despite treatment, but it requires further validation in longitudinal studies.

全身性エリテマトーデスにおける抗ヌクレオソーム抗体の意義 : 臨床・病理組織学的研究

全身性エリテマトーデスにおける抗ヌクレオソーム抗体の意義 : 臨床・病理組織学的研究

Anti-DNA and Anti-Nucleosome Antibodies: An Update

Among the large spectrum of autoantibodies (Ab) detected in systemic lupus erythematosus (SLE), anti-dsDNA Ab have been used to diagnose and assess disease activity for more than 60 years. Anti-dsDNA Ab are often associated with anti-nucleosome Ab, and in clinical practice combining both is helpful for diagnosis, prognosis, and as biomarkers to monitor response to treatment, especially when using drugs that target B cells and Ab production. In general, the association of anti-dsDNA Ab plus anti-nucleosome Ab is linked with disease flares and can indicate lupus nephritis. Moreover and since anti-nucleosome Ab have higher sensitivity and specificity than anti-dsDNA Ab, the former is a useful marker in SLE patients negative for anti-dsDNA Ab and in the diagnosis of certain types of drug-induced lupus.

Semaphorin 5A, Anti-Nucleosome Antibodies and Ferritin: Disease Activity Markers in Patients with Systemic Lupus Erythematosus

Background: Systemic Lupus Erythematosus (SLE) is a multi-factorial, chronic autoimmune disorder characterized by dysfunction of T and B lymphocytes. Although the prognosis of SLE has improved in the past few decades, the absence of biomarkers for residual activity in various organs and the early detection of disease flares hamper further management. Conventional serologic markers of SLE, such as anti-dsDNA and complement levels, are not ideal, as they are not sufficiently sensitive and specific for the diagnosis of the disease and monitoring of disease activity. Thus, novel biomarkers for SLE activity have to be developed. Semaphorin 5A (Sema 5A), antinuclosome antibody (Anu A) and ferritin may fall into this category of novel bio markers.Objective: To evaluate the role of serum Sema 5A, Anu A and ferritin in detection of SLE activity.Methods: The present study included 40 patients with SLE divided according to Systemic Lupus Erythematous Disease Activity Index (SLEDAI) into two groups: 20 active SLE and 20 inactive SLE patients. They were compared with 20, sex and age matched healthy individuals for control. Levels of Sema 5A, Anu A were measured by Enzyme- Linked Immunosorbent Assay (ELISA) and serum ferritin levels were measured by Electrochemiluminescence immunoassay (ECLIA).Results: There was highly significant increase in Sema 5A, Anu A and ferritin levels inactive when compared within active SLE patients and control subjects (P=0.000 and P=0.000 and P=0.000 respectively). There was a significant increase in ferritin level in inactive SLE patients when compared with control subjects (P=0.045) with no significant difference regarding Sema 5A, Anu A (P3=0.089 and 0.225 respectively). There were positive correlations between Sema 5A, Anu A, ferritin and each of SLEDAI, and CRP. Positive correlation also found between Sema 5A and each of Anu A and ferritin and between AnuA and ferritin. Negative correlation was found between Sema 5A and C3 and between Anu A, ferritin and C4. Significant relation was found between Sema 5A, Anu A, ferritin and each of ANA and anti-dsDNA in active SLE patients. Area under the curve (AUC) for ferritin, Sema 5A and Anu A were (0.861, 1.0 and 1.0 respectively).Conclusion: Our study showed that the serum proteins semaphorin 5A, antinuclosome antibodies and ferritin may be useful markers in monitoring disease activity in patients with SLE. Regarding area under the curve (AUC), these serum protein markers result in even better sensitivity and specificity profiles in picking up early relapse of SLE.

Evaluating Anti-SmD1-amino-acid 83-119 Peptide Reactivity in Children with Systemic Lupus Erythematosus and Other Immunological Diseases.

Background:SmD1-amino-acid 83-119 peptide (SmD183-119) is the major epitope of Smith (Sm) antigen, which is specific for adult systemic lupus erythematosus (SLE). The anti-SmD183-119 antibody has exhibited higher sensitivity and specificity than anti-Sm antibody in diagnosing adult SLE. However, the utility of anti-SmD183-119 antibodies remains unclear in children with SLE (cSLE). This study aimed to assess the characteristics of anti-SmD183-119 antibody in the diagnosis of cSLE.Methods:Samples from 242 children with different rheumatological and immunological disorders, including autoimmune diseases (SLE [n = 46] and ankylosing spondylitis [AS, n = 11]), nonautoimmune diseases (Henoch-Schonlein purpura [HSP, n = 60], idiopathic thrombocytopenia purpura [n = 27], hematuria [n = 59], and arthralgia [n = 39]) were collected from Shanghai Children's Medical Center from March 6, 2012 to February 27, 2014. Seventy age- and sex-matched patients were enrolled in this study as the negative controls. All the patients' sera were analyzed for the anti-SmD183-119, anti-Sm, anti-U1-nRNP, anti-double-stranded DNA (dsDNA), anti-nucleosome, anti-SSA/Ro60, anti-SSA/Ro52, anti-SSB, anti-Scl-70, and anti-histone antibodies using the immunoblotting assay. The differences in sensitivity and specificity between anti-SmD183-119 and anti-Sm antibodies were compared by Chi-square test. The correlations between anti-SmD183-119 and other auto-antibodies were analyzed using the Spearman's correlation analysis. A value of P < 0.05 was considered statistically significant.Results:Thirty-six out of 46 patients with cSLE were found to be positive for anti-SmD183-119, while 12 patients from the cSLE cohort were found to be positive for anti-Sm. Compared to cSLE, it has been shown that anti-SmD183-119 was only detected in 27.3% of patients with AS and 16.7% of patients with HSP. In comparison with anti-Sm, it has been demonstrated that anti-SmD183-119 had a higher sensitivity (78.3% vs. 26.1%, χ2 = 25.1, P < 0.05) and a lower specificity (90.8% vs. 100%, χ2 = 13.6, P < 0.05) in the diagnosis of cSLE. Further analysis revealed that anti-SmD183-119 antibodies were positively correlated with anti-dsDNA, anti-nucleosome, and anti-histone antibodies in cSLE. Moreover, it has been clearly shown that anti-SmD183-119 was more sensitive than anti-Sm in discriminating autoimmune diseases from nonautoimmune disorders in patients with arthralgia or hematuria.Conclusions:Measurement of anti-SmD183-119 in patients with cSLE has a higher sensitivity and a marginally lower specificity than anti-Sm. It has been suggested that inclusion of anti-SmD183-119 testing in the integrated laboratory diagnosis of cSLE may significantly improve the overall sensitivity in child populations.

Antibodies against antigens related to scleroderma in a cohort of patients with morphea.

Morphea is a rare fibrosing skin disorder. Antinuclear antibodies (ANA), anti-histone and rheumatoid factor are detected in high rate of morphea cases. Scleroderma-related antibodies are usually absent. Twenty-one adult patients affected by morphea were examined at the time of enrollment and after 6 months with the assessment of clinical outcome measures of disease severity and damage. Healthy subjects were considered as normal controls while patients affected by systemic sclerosis and other connective tissue diseases (CTD) were considered as pathological controls. Serum samples from all the patients and controls were analyzed for the detection of ANA, anti-nucleosome antibodies, anti-dsDNA and anti-extractable nuclear antigen. Scleroderma-related autoantibodies were searched using a line-blot test. We enrolled 21 patients affected by morphea. ANA were found in 12 patients (57%). Anti-DNA and anti-nucleosome antibodies were negative in all cases. Systemic sclerosis-specific antibodies were found in 11 morphea patients (52.4%) and one healthy control (6.25%). In patients affected by systemic sclerosis (100%) and different CTD (%), scleroderma-related autoantibodies were more frequently detected than in morphea (52.4%). In morphea, anti-TRIM21/Ro52 antibodies were found in 4 patients (36.4%) and resulted to be the most frequently detected antibodies also in two groups of SSC and CTD. The high prevalence of ANA and the identification of some antagonists of systemic sclerosis-related autoantibodies, confirm the idea of a significant activation of autoimmune system in morphea.

Change in autoantibody and cytokine responses during the evolution of neuromyelitis optica in patients with systemic lupus erythematosus: A preliminary study

Background: Neuromyelitis optica (NMO)–systemic lupus erythematosus (SLE) association is a rare condition characterized by multiple autoantibodies. Objective: To examine if, during the evolution of NMO, anti-AQP4 responses are part of polyclonal B cell activation, and if T cell responses contribute. Methods: In 19 samples of six patients who developed NMO during SLE, we examined the correlation of AQP4-IgG1 and IgM with (i) anti-MOG IgG and IgM, (ii) anti-nuclear, anti-nucleosome and anti-dsDNA IgG antibodies, (iii) cytokines and chemokines in the serum and (iv) longitudinal relation to NMO relapses/remission. Results: AQP4-IgG1 was present 1–2–5 years before the first NMO relapse. During relapse, AQP4-IgG1, ANA, anti-dsDNA and anti-nucleosome antibodies were elevated. Anti-MOG IgG/IgM and AQP4-IgM antibodies were not detected. AQP4-IgG1 antibodies correlated with concentration of anti-nucleosome, IFN-γ,interferon-gamma-induced CCL10/IP-10 and CCL17/TARC (p<0.05, respectively). CCL17/TARC correlated with levels of anti-nucleosome and anti-dsDNA (p<0.05, respectively). Compared to healthy subjects, concentration of IFN-γ and CCL17/TARC was higher in NMO/SLE (p<0.05). Conclusions: AQP4-IgG1 antibodies are present in the sera years before the first NMO attack in patients with SLE; elevation of anti-AQP4 is part of a polyclonal B cell response during NMO relapses; in spite of multiple autoantibodies in the serum, MOG antibodies were not present; Th1 responses accompany autoantibody responses in NMO/SLE.

Abstract 4476: Epigenetically altered circulating nucleosomes as blood biomarkers for early detection of cancer: clinical studies in NSCLC, CRC, PCA and PC

Abstract BACKGROUND: The epigenetic structure of chromosomes in cancer tissue is altered with respect to histone modification, histone isoform and DNA modification composition as well as the composition of non-histone chromatin proteins - for example; the androgen receptor (AR) in prostate cancer. Short (&amp;gt;180bp) fragments of tumor-derived cell free DNA circulate in the blood of cancer patients as mono-nucleosomes. AIM: To evaluate whether epigenetically altered circulating nucleosomes provide a blood biomarker opportunity for early detection of cancers. METHODS: We have developed more than 25 ELISA tests for circulating nucleosomes containing different epigenetic signals. These assays employ a solid phase anti-nucleosome antibody in combination with a labelled antibody directed to bind to a particular epigenetic signal - for example a histone modification, histone isoform, modified cytosine residue or a nucleosome-AR adduct. We have conducted clinical studies in collaboration with a number of centres to investigate the use of these ELISA tests for the detection of epigenetically altered circulating nucleosomes in serum samples taken from patients with lung cancer, colorectal cancer, pancreatic cancer and prostate cancer and from control subjects with no cancer and with or without concomitant disease of the same organ. RESULTS: A collaborative study of 121 patients referred for colonoscopy at the CHU Dinant Godinne - UCL Namur, Belgium, who either showed potential symptoms of CRC or were high-risk subjects found that a panel test of five epigenetic nucleosome biomarker assays detected 87% of colorectal cancer cases and 67% of dysplastic polyps at 90% specificity. A 59 subject collaborative study with Lund University, Sweden, including 25 patients with operable stage 2 pancreatic cancer, 10 patients with other pancreatic diseases and 24 healthy individuals found that a panel of 5 assays including 4 epigenetic nucleosome ELISA assays and CA19-9 detected 92% of early-stage pancreatic cancer cases, with 100% specificity among the healthy subjects and 2 false positives among patients with benign pancreatic disease. A study of 73 patients conducted with the Liege University Hospital, Belgium, including 29 NSCLC cases, 22 COPD cases and 22 subjects healthy lungs showed that a panel of 4 epigenetic nucleosome assays combined with the smoking history detected 93% of NSCLC cases, with 91% specificity and also differentiated NSCLC and COPD cases. Collaborative studies in prostate cancer have also shown excellent results and data from these studies will be presented at the conference. CONCLUSION: Epigenetic profiling of circulating nucleosomes using simple ELISA methods offers a biomarker opportunity for the early detection of a variety cancers. Citation Format: Jake V. Micallef. Epigenetically altered circulating nucleosomes as blood biomarkers for early detection of cancer: clinical studies in NSCLC, CRC, PCA and PC. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4476.

Interleukin‐6 receptor alpha blockade improves skin lesions in a murine model of systemic lupus erythematosus

Systemic lupus erythematosus (SLE) is an autoimmune disease, characterized by antinuclear autoantibodies (ANA) and immunocomplexes, commonly affecting kidneys, skin, heart, lung or even the brain. We have shown that JunBΔep mice develop a SLE phenotype linked to increased epidermal Interleukin (IL)‐6 secretion. Blocking of IL‐6 receptor alpha (IL‐6Rα) is considered as therapeutic strategy for the treatment of SLE. JunBΔep and wild‐type mice were treated for short (5 weeks) or long term (21 weeks) with the IL‐6Rα‐blocking antibody MR16‐1. Skin and kidney of mice were investigated by histology and immunofluorescence, and in addition, kidneys were analysed by electron microscopy. Furthermore, soluble IL‐6R (sIL‐6R), antihistone and antinucleosome antibodies levels were measured and associated with disease parameters. Treatment with MR16‐1 resulted in significant improvement of SLE‐like skin lesions in JunBΔep mice, compared to untreated mice. The sIL‐6R amount upon long‐term treatment with MR16‐1 was significantly higher in JunBΔep versus untreated JunBΔep (P = 0.034) or wild‐type mice (P = 0.034). MR16‐1 treatment over these time spans did not significantly improve kidney pathology of immunoglobulin deposits causing impaired function. Significantly higher antihistone (P = 0.028) and antinucleosome antibody levels (P = 0.028) were measured in MR16‐1‐treated JunBΔep mice after treatment compared to levels before therapy. In conclusion, blockade of IL‐6Rα improves skin lesions in a murine SLE model, but does not have a beneficial effect on autoimmune‐mediated kidney pathology. Inhibition of IL‐6R signalling might be helpful in lupus cases with predominant skin involvement, but combinatorial treatment might be required to restrain autoantibodies.

Clinical analysis of 6 patients with drug-induced lupus

To improve the understanding of drug-induced lupus (DIL) and the differences from systemic lupus erythematosus (SLE). Clinical manifestation and treatment of patients with definite DIL were retrospectively analyzed. Six patients with DIL were enrolled in this study, including 4 females and 2 males. Two patients were diagnosed after receiving interferon, one after soluble tumor necrosis factor receptor fusion protein, one after propylthiouracil, one after penicillamine, and one after levofloxacin. High titer of antinuclear antibody was identified in all six patients, including 3 with positive anti-dsDNA antibody. One patient had positive anti-Sm antibody. One patient had positive anti-RNP antibody. One patient had anti-nucleosome antibody. One patient had anti-histone antibody. One patient had antimitochondrial antibodies-M2, and one patient had anticardiolipin antibodies. Patients with DIL are not as severe as those with SLE. After cessation of suspected drugs and administration of standard treatment, the clinical outcome of DIL is satisfying.

Indirect immunofluorescence on different cell line in detection of cell membrane DNA antibody in juvenile systemic lupus erythematosus

Objective To detect anti-cell membrane DNA(cmDNA)antibody with human B lymphocyte Raji cells and human promyelocytic leukemia HL60 cells as substrates and to compare the diagnostic value of anti-cmDNA antibody with that of anti-nucleosome antibody(AnuA), anti-Sm antibody and anti-double-stranded DNA(dsDNA)antibody in juvenile systemic lupus erythematosus(JSLE)patients. Methods We recruited 92 JSLE patients and 71 patients with other rheumatic diseases.Anti-cmDNA antibody andantinuclear antibody(ANA)was detected in patient serum by indirect immunofluorescence assays(IIF). Anti-dsDNA antibody were detected by combining enzyme-linked immuno sorbent assay(ELISA)and IIF.Anti-Sm antibody were detected by double immunodiffusion assay and immunoblotting, while anti-nucleosome antibody(AnuA)were detected by ELISA.We collected concurrent clinical data. Results Anti-cmDNA antibody demonstrated stronger intensity of fluorescent patterns in using Raji cells as substrate than HL60 cells.JSLE patients had a significantly higher positive percentage of anti-cmDNA than patients with other rheumatoid diseases.The sensitivity of anti-cmDNA on cell line Raji was higher than that of anti-dsDNA and anti-Sm(P 0.05)and was lower than anti-Sm and AnuA(P 0.05)and the specificity was lower than AnuA(P<0.01). The sensitivities of anti-dsDNA, anti-Sm and AnuA by combining with anti-cmDNA were much higher than that of the above antibody detected respectively(P<0.05). Anti-cmDNA had no correlation with SLE disease activity index(P=0.907). Conclusion The high sensitivity and specificity of anti-cmDNA antibody make it a valuable diagnostic tool for JSLE.Combined detection of anti-cmDNA and other autoantibody might further improve the sensitivity in JSLE.Anti-cmDNA detected with IIF on cell line Raji was better than cell line HL60. Key words: Systemic lupus erythematosus; Child; Diagnosis; Anti-cmDNA antibody

Clinicopathologic Characteristics and Outcomes of Lupus Nephritis With Antineutrophil Cytoplasmic Antibody: A Retrospective Study.

Few studies have analyzed the clinicopathologic characteristics and outcomes of lupus nephritis (LN) patients with antineutrophil cytoplasmic antibody (ANCA).The clinical and renal histopathologic data of 154 patients with biopsy-proven LN from 2011 to 2013 were analyzed retrospectively. The patients were followed up for a median period of 16.8 ± 9.4 months, and their outcomes were analyzed. Multivariate Cox analysis was used to evaluate the independent factors for poor outcomes.Among the 154 LN patients, 26 (16.88%) were seropositive for ANCA. The incidences of alopecia, oral ulcer, photosensitivity and skin lesion, and psychosomatic manifestations in the ANCA-positive group were significantly higher than in the ANCA-negative group (P = 0.007, 0.02, 0.02, and 0.03, respectively). Compared with the ANCA-negative group, the ANCA-positive group had significantly lower levels of complement C3 (P = 0.03). Additionally, the positive rate of antinucleosome antibodies, antihistone antibodies, antimitochondrial antibody M2, and anticardiolipin antibodies were higher significantly in the ANCA-positive patients than in the ANCA-negative patients (P = 0.001, 0.001, 0.03, 0.005, respectively). The ANCA-positive group had a notably higher chronic index than the ANCA-negative group (P = 0.01). During the follow-up, the complete remission rate in the ANCA-negative group was higher than that in the ANCA-positive group (P = 0.01). The cumulative renal survival rate in the ANCA-positive group was significantly lower than in the ANCA-negative group (log-rank = 6.59, P = 0.01). Multivariate Cox analysis revealed that the reduced estimated glomerular filtration rate (HR, 1.02; 95% confidence interval, 1.01 to 1.03; P = 0.005), NLR (HR, 1.20; 95% confidence interval, 1.02 to 1.40; P = 0.03), and ANCA (HR, 3.37; 95% confidence interval, 1.12 to 10.09; P = 0.03) were independent risk factors for patients’ renal survival after adjusting for age, sex, crescent formation, and glomerulosclerosis.The study found ANCA in LN patients is not rare, and patients with ANCA present with more severe clinicopathologic injuries. Thus, ANCA is an independent risk factor for poor renal outcomes in LN patients.