Search

Background: Dense fine speckled (DFS) pattern is defined by very intense, heterogeneous speckled staining of nucleoplasms of interphase HEp-2 cells and chromosomal areas of metaphase cells. The association of Anti-DFS70 and rheumatologic signs, symptoms, and diagnosis were evaluated. Methods: One-hundred-eight anti-DFS70 positives who got consecutively admitted to the Rheumatology clinic between January and June 2020 were analyzed. The clinical and laboratory findings of positives for anti-DFS70 antibody were compared with those with DFS pattern ANA IFA staining rates. Also, anti-DFS70 positivity rates and their correlation with the DFS staining pattern were analyzed retrospectively in 1016 CTD patients. Results: The most common complaint was joint pain seen in 77 (71.3%) and the most common laboratory abnormality was RF-positivity observed in 10/108 (9.3%) who had anti-DFS70 positivity. The most common ANA staining pattern was DFS (72/108; 66.7%); one-third had other than DFS. No statistical significance was found for the association of any of the rheumatological complaints and laboratory findings with the DFS staining pattern. ANA analysis was performed in a total of 964/1016 (94.88%) CTD patients and 44 (4.56%) of these positive for anti-DFS70. The correlation coefficient showed good correlations between the DFS pattern staining and anti-DFS70 antibody positivity (r=+0.773, p Conclusions: Anti-DFS70-positives have a low rate of CTD. A low anti-DFS70 positivity rate was observed in patients with CTD. As such, it can be considered that anti-DFS70 does not predict CTD or even excludes it.

Autoantibody assays are reported in a variety of formats. Results only slightly above established cut-offs provide lower likelihood ratios; therefore, their clinical significance may be more uncertain, which is not readily communicated with dichotomous qualitative reporting. Line immunoassays (LIA) are a common method for detecting antibodies to extractable nuclear antigens (ENA) and myositis-associated antibodies. However, recommended positive cut-offs are contentious. We distributed a survey via e-mail to participants in the Royal College of Pathologists of Australasia Quality Assurance Program (RCPAQAP) Immunology modules and to a dedicated immunology mailing list in Australasia. Questions explored general viewpoints surrounding autoantibody reporting, as well as current laboratory practices, with particular focus on interpretation and reporting of the most commonly used ENA LIA manufactured by Euroimmun. There were 31 responders, representative of at least 17 unique laboratories across Australia (8 public, 5 private) and New Zealand (4 laboratories). Responses suggest that autoantibody reporting is not standardised; there was variation in general viewpoints and reporting practices, particularly regarding the interpretation of and positive cut-offs used for the Euroimmun ENA LIA, which were contrary to the manufacturer's guidelines in a majority of the responses. Interpretative qualitative reporting based on results from other investigations and the clinical history was a common theme. There is large variation in the reporting of autoantibody assays within Australasia, especially by LIA. A majority of respondents report the most commonly used ENA LIA contrary to manufacturer's guidelines; alternative positive cut-offs are commonly utilised. LIA reports should indicate the level of positivity to enhance their relevance in the clinical decision-making process.

The current study aimed to characterize patients from a rheumatology referral center in north India, who satisfied the definition of interstitial pneumonia with autoimmune features (IPAF) as given by the American Thoracic Society and European Respiratory Society (ATS/ERS) consensus committee in 2015. Thirty-five adult patients aged 18 years and above, fulfilling the 2015 ATS/ERS criteria for IPAF were included in the study. The clinical and immunological profile, and radiologic findings on high-resolution computerized tomography thorax were noted. Antinuclear antibody (ANA) by indirect immunofluorescence at 1:320 titer and myositis-specific antibody (MSA) assays were performed. Non-parametric tests were used to compare variables between groups. The study cohort included predominantly female patients with a mean age of 50.6 ± 13 years and mean duration of disease of 38.8 ± 28.4 months. Majority of patients (49%) fulfilled the morphologic and serologic domains as per the IPAF consensus criteria and 31% patients had features in all three domains. Non-specific interstitial pneumonia was the most common pattern observed in 77% patients. Raynaud’s phenomenon and inflammatory arthritis were the predominant autoimmune features. Pulmonary arterial hypertension was documented in 60% of patients on echocardiography. Positive ANA at 1:320 dilution was present in all 26 patients tested, whereas extractable nuclear antigen and MSA assays detected autoantibodies in 49% and 51% of patients respectively. IPAF predominantly affected females in the age group of 50 years and above, with varied autoimmune manifestations and autoantibody profile. Supplementary Information The online version contains supplementary material available at 10.1007/s00296-021-04883-7.

Introduction Serum autoantibodies support the diagnosis of interstitial lung disease (ILD) related to systemic autoimmune diseases (SAD-ILD). Nevertheless, their presence in the bronchoalveolar lavage (BAL) has not been explored. Objectives To demonstrate the presence of autoantibodies in the BAL of ILD patients at onset of clinical evaluation, its relation with serum autoantibodies and to analyze clinical features of patients with autoantibodies in BAL. Methods Autoantibodies against extractable nuclear antigens (ENAs) were analyzed by immunoblot in the BAL of 155 patient with suspected diagnosis of ILD and 10 controls. Results Seven ENAs were detected in the BAL of 19 patients (Anti-Ro52, Anti-Ro60, CENP-B, Anti-La, Jo-1, Sm/RNP and Anti-SL70). The most frequent ENA was anti-Ro52 (13 patients; 68,4% of positives ones). Seven patients presented more than one ENAs. Fourteen were diagnosed of SAD-ILD, 3 of interstitial pneumonia with autoimmune features, one of non-specific idiopathic pneumonia and other of silicosis. In 10 cases (52%) IgA autoantibodies were also detected. The autoantibodies observed in BAL were also detected in the serum of 17 patients (90%). There were no significant clinical differences with the patients with SAD-ILD or interstitial pneumonia with autoimmune features with patients with negative BAL. Conclusion The study of ENAs in BAL is feasible and can be a useful tool in the ILD initial algorithm, specifically sustaining the suspected diagnosis of SAD-ILD.

Objectives To study the frequency of different autoantibodies to extractable nuclear antigens (ENAs) in rheumatoid arthritis (RA) patients and to correlate findings with clinical manifestations, disease activity and radiological damage. Methods A total of 230 RA patients were included and 75 healthy controls. In all patients rheumatological assessment was done and routine laboratory investigations and immune profile were performed in both patients and controls, including: RF, ACPA, ANA and anti-ENAs (Ro/SSA, La/SSB, U1-RNP, anti-Jo-1 and anti-Sm). Radiological damage was scored using Sharp/van der Heijde, and disease activity was evaluated by DAS28-ESR and DAS28-CRP. Results RF was positive in 101 (43.9%), ACPA in 220 (95.7%), ANA in 58 (25.2%), anti Ro in 31 (13.5%), anti-La in 10 (4.3%), anti-Jo1 in 5 (2.2%) and anti-RNP in 2 (0.9%). Anti-Ro/SSA positively correlated with sicca symptoms (p = .02), RF titer (p < .001), ANA (p < .001), DAS28-ESR (p = .026), and DAS28-CRP (p = .003). Anti-La antibodies correlated positively with SJC (p = .001), TJC (p = .001), ANA (p < .001), DAS-28 ESR (p = .007). Anti-Jo-1 correlated positively with interstitial lung disease (ILD) (p ≤ .001), RF titer (p = .037) and ANA (p ≤ .001). Anti-RNP antibodies correlated positively with disease duration (p ≤ .001), ACPA titer (p ≤ .001) and ANA (p = .014). In the controls ANA was positive in two (2.7%), anti-Ro in three (4%), and none of the controls tested positive for other autoantibodies. Conclusions In RA patients, positive ANA is frequent and positively associated with anti-Ro, anti-La and anti-Jo1 autoantibodies. Screening for autoantibodies against other anti-ENAs seems mandatory in RA patients especially when ANA is positive. RA cases with positive Anti-Jo-1 may develop anti synthetase syndrome and ILD.

Summary Background Acute viral myositis (AVM) may be triggered by influenza A/B, enteroviruses and other viruses. Severe complications including rhabdomyolysis regularly lead to acute kidney injury (AKI). The aim of this short report was to discuss management and differential diagnosis of massive creatine kinase (CK) elevation. Patient, material and methods Herein, we report on a 19-year-old Austrian male of African descent with a history of respiratory tract infections and whole-body pain. He further developed acute viral myositis and massive CK elevation up to 440,000 IU/L but without any signs of AKI. A literature search relating AVM, management and differential diagnosis of rhabdomyolysis was conducted in PubMed and UptoDate. Results A full panel of serological and autoimmune blood work-up including testing for human immunodeficiency virus (HIV), hepatitis, influenza A/B, Epstein-Barr virus (EBV), antinuclear antibodies (ANA) and autoantibodies against various extractable nuclear antigens (ENA) did not reveal evidence for viral originators or autoimmune diseases. This case indicates that in acute viral myositis associated with extreme CK elevation (>400,000 IU/L) AKI might be completely absent. Potential causes for this clinical phenotype, differential diagnosis and management are discussed.

Individuals with inflammatory rheumatic diseases such as lupus are considered at risk for severe COVID-19 infection due to their immunocompromised state and the use of immunosuppressive treatment. Its impact on lupus is yet to be determined. Herein, we report a case of COVID-19 pneumonia in a 40-year-old female with a history of hypothyroidism and mild pulmonary hypertension who presented with a 4-day history of fever and breathlessness. COVID-19 infection was confirmed by real-time polymerase chain reaction (RT-PCR). On examination, she had a malar rash, short hair, and crepitations on the right side infrascapular and infraaxillary areas. Investigations revealed to have anemia, thrombocytopenia, transaminitis, high ferritin, and low fibrinogen, which were suggestive of cytokine release syndrome. Her antinuclear antibody (ANA) by indirect immunofluorescence on Hep-2 cells was 4+ homogeneous in 1:100 titer dilution and extractable nuclear antigen panel showed antibodies positive for histone, nucleosome, Ro, and smith antigens. The patient was diagnosed with systemic lupus erythematosus (SLE) with severe COVID-19 pneumonia with cytokine release syndrome and managed conservatively without specific antivirals or steroids. Her clinical condition and laboratory parameters improved afterward. On the 12th day, her RT-PCR for COVID-19 was negative and the patient was discharged. This would probably a reference case which showed conservative management can be helpful at times where no definite antiviral therapy has been approved with judicious use of immunosuppressants. Close monitoring of clinical conditions and laboratory parameters are recommended for such autoimmune disorders with COVID-19 infection.

Background Autoimmunity causes the loss of normal immune homeostasis and involves the presence of autoantibodies and inflammation. Thromboangiitis obliterans or Buerger's disease (BD) refers to a type of vascular obstructive syndrome, with tobacco exposure accounting for disease formation and progression. However, the current understanding of autoimmunity is unclear in the context of BD, and the scientific findings are not enough to support autoimmune mechanisms. This study was aimed at investigating autoimmunity factors in patients with BD. Methods Clinical and experimental examinations were performed on 80 patients with BD. The diagnostic work-up for autoimmunity was composed of IgM rheumatoid factor (RF), anti-nuclear antibodies (ANA), The erythrocyte sedimentation rate (ESR), anti-cyclic citrullinated peptide (CCP) antibodies, Antiphospholipid antibodies (APA), Anti-cardiolipin antibodies (ACLA), anti-double-stranded DNA (ds-DNA), and extractable nuclear antigen (ENA) profile. Immunomarkers were detected using the quantitative enzyme-linked immunosorbent assay (ELISA). Results Raynaud's phenomenon (84.93%), cold sensitivity (76.25%), and claudication (73.75%) were the most common symptoms in the BD patients. Also, 64.29% represented with high ANA levels and positive RF, while 42.11% were found with increased ANA and ESR levels. The ANA/RF positive BD patients had ESR> 15 mm/hr and a high prevalence of cold sensitivity, claudication, and Raynaud's phenomenon (p> 0.05). Conclusion There is a possibility of a non-specific autoimmune disposition among BD patients. RF and ANA could be considered for predicting disease progression.

The incidence of autoimmune disorders that includes the connective tissue diseases has seen a rise in India in recent times. Antinuclear antibodies, the telltale sign of systemic autoimmune response, thus can be used as a screening tool and also to support the diagnosis of systemic autoimmune disease. The present retrospective cross- sectional analysis aimed to study the antinuclear antibodies profile (patterns and specific antibody reactivity) amongst suspected cases of auto-immune disorders at a tertiary care teaching hospital. The study retrieved and reviewed reports of 644 patients sent for ANA testing by indirect immunofluorescence assay over a period of 1 year by different specialty departments. Positive samples were further processed for anti-ds-DNA antibody and antibodies to extractable nuclear antigen. Data collected was statistically analysed. ANA pattern positivity was observed in 31% of cases and a positive antibody reactivity was seen in 66% of them. Female predominance (82%) was noted in both pattern positivity and antibody reactivity. High levels of pattern positivity and antibody reactivity was found in the young adults (45.9%). Amongst the ANA patterns, the nuclear homogenous pattern was found the commonest. The common antibodies associated with this pattern were anti-dsDNA and U1 Sm/RNP antibodies. A stronger fluorescence intensity on initial screening showed a higher confirmation rate for specific antibodies on immunoassay. High occurrence of positive ANA patterns in autoimmune disorders suggests its utilization as a screening tool for them and would also play an adjuvant to the diagnosis. Early knowledge about future autoimmunity will earn better prognostic achievements through better treatment interventions.

A 52-year-old male patient developed RA in March 2009 at the age of 43, with symmetric polyarthritis and active synovitis affecting hands, knees, ankles and both feet without symptoms or signs suggestive of extra-articular features. Laboratory investigations showed negative RF and positive anti-CCP antibodies, negative ANA, negative anti-dsDNA antibodies; the X-rays of both hands showed typical erosive changes in RA and fulfilled the new ACR/EULAR (2010) criteria of RA. The patient achieved remission on a combination of DMARDs. He did well until January 2017 when he developed acute onset of progressive chest pain, dyspnea, and acute respiratory failure. High-resolution CT of the lung showed extensive areas of ground glass veiling, and interstitial subpleural infiltrates were found consistent with aggressive interstitial lung disease (ILD). Autoantibodies against extractable nuclear antigens were screened and showed positive results for anti-RO and anti-Jo1 autoantibodies. The positive anti-Jo1was an expression of anti-synthetase syndrome complicating the RA course and explained the rapidly aggressive course of ILD.

Point-of-care testing (POCT) is a near-patient detection technique that could provide results within minutes. Compared to conventional laboratory testing, POCT is more beneficial particularly in emergency and urgent care settings. Detection of antinuclear antibodies and autoantibodies against extractable nuclear antigens are critical for the diagnosis and management of autoimmune rheumatic diseases (AIRD). At present, these autoantibodies are detected using conventional methods that take hours or days in centralized laboratories. POCT for AIRD is hardly practiced as they are chronic illnesses which are seen as clinical emergencies occasionally. Since POCT provides instant results, it may be life-saving in critically ill and confusing cases, particularly those admitted to intensive care units with multisystem organ failure. POCT is also very useful in rural and remote health-care centers, addressing the needs of emergent disease detection. This review aims to provide an overview of the potential use of POCT in AIRD as well as discuss the types, benefits, and shortcomings of POCT devices, and the hurdles that prevent their widespread clinical use.

Antinuclear antibodies (ANA) and extractable nuclear antigens (ENA) are instrumental biomarkers crucial in the detection of autoimmune disorders (AID) such as systemic lupus erythematosus (SLE), Sjogren syndrome, etc.. In the present study, an assessment of the most frequent ANA patterns associated with most detectable ENA that could be used as efficient prognostic markers in the diagnosis of autoimmune diseases was conducted. Data was retrospectively analyzed from AID patients, retrieved from the medical records of King Fahad Medical City, Riyadh, KSA, from January 2016 to October 2018 who underwent ANA immunofluorescence of HEp-2 cells and their ENA detection was studied. Of the 453 total patients, 39/55 AID males (71%) and 332/398 AID females (83.4%) exhibited ANA positivity. The most common pattern was speckled S-ANA (32.4%) in females and homogenous H-ANA pattern (25.4%) in males. The histones were found at higher frequency in different ANA patterns. anti-Sjogren syndrome related antigen A (SSA), anti-ribonucleoprotein antibody (RNP-Sm), and histones were observed to be associated with homogenous and speckled nuclear patterns. Frequencies of ENA in all ANA patterns were found significant at p < 0.05 in males and p < 0.001 in females. Spearman's rank correlation of ENA within and among the ANA patterns was non-significant. SSA was significantly correlated with RNP-Sm and Sm at p < 0.05 and p < 0.01, respectively. The extractable nuclear antigens SSA, RNP-Sm, and histones were found associated with the S-ANA and H-ANA patterns. These correlations are of relevance for the accurate diagnosis of autoimmune diseases.

The treatment of Buerger's disease (BD) presents a medical problem as its etiology is still unclear. In this study, our objective was to evaluate the serum levels of autoimmune markers in patients with different clinical features of BD.In this study, 80 BD patients were categorized in three groups using a cross-sectional design: migratory thrombophlebitis, cold sensitivity, and skin discoloration (mild symptoms); chronic ulcers, claudication, and burning pain of the feet at night (moderate symptoms); pain at rest and spontaneous gangrene (severe symptoms). Enzyme-linked immunosorbent assay was performed to measure antibodies against immunoglobulin M rheumatoid factor (IgM RF), anti-nuclear antibodies (ANA), antibodies against cyclic citrullinated peptide (anti-CCP), antiphospholipid antibodies (APA), anti-cardiolipin antibodies (ACLA), anti-double stranded DNA (anti-dsDNA), and extractable nuclear antigen (ENA) profile.Patients with severe symptoms showed the lowest age (p=0.031), ESR (p0.001), and highest prevalence of ischemia (p0.001). In all the patients, the serum levels of ANA and IgM RF were higher than 1 U and 15 IU/mL, respectively. However, the progression of the disease from mild to moderate did not affect these markers significantly (p0.05). Other markers were negative in patients with BD.The findings of this study indicate that BD may closely be correlated to transient autoimmune phenomena, despite the fact that further research is required to investigate how transient unspecific autoimmune reactions contribute to the BD pathogenesis.

Markers of autoimmunity, such as autoantibodies, have been found in patients with endometriosis. These include the antinuclear antibodies (ANA). We aimed to evaluate the prevalence of ANA in a sample of patients with endometriosis and its possible clinical associations. Ninety-four patients with endometriosis and 91 controls were studied for ANA and extractable nuclear antigen (ENA; anti-Ro, anti-La, anti-Sm, and anti-RNP) profiles and anti-dsDNA. Epidemiological, clinical, and staging data in endometriosis were obtained. Patients with autoimmune disorders were excluded. Patients with endometriosis had a 21.2% prevalence of positive ANA vs. 5.4% in controls (P = 0.001). The ENA profile and anti-dsDNA were negative. Patients with positive ANA were more asymptomatic (P = 0.03) and had less dysmenorrhea. No associations with disease duration, patient age, or endometriosis stage were found. We found a high prevalence of positive ANA in patients with endometriosis. The presence of this autoantibody may be linked to a milder clinical expression of the disease.

BACKGROUND Lupus nephritis (LN) is common feature of Systemic Lupus Erythematosus (SLE) and affects 50% of patients with SLE. Racial differences in incidence and prevalence have been well documented worldwide. In Australia, higher incidence and prevalence of SLE had been previously reported in Aboriginal and Torres Strait Australians compared to non-Indigenous Australians. AIM to describe the differences in clinical features and lupus biomarkers between Aboriginal and Torres Strait Islander Australian and non-Indigenous Australian LN patients. METHODS We retrospectively identified all consecutive biopsy-proven LN patients in our institution and compared the clinical features and lupus biomarkers between Aboriginal and Torres Strait Islander Australians and non-indigenous Australians. RESULTS Of the thirty-three consecutive biopsy proven LN patients, twenty-six self-identified as of Aboriginal and Torres Strait Islander descent. The estimated incidence of lupus nephritis in Aboriginal and Torres Strait Islander Australian and non-Indigenous Australians were 5.08 and 0.47 per 100,000 patient years respectively. Neurological manifestations (23.08% vs 0%), haematological manifestations (46.50 % vs 16.67) and right heart catheter proven pulmonary arterial hypertension (23.08% vs 0%) were more frequently observed amongst Indigenous Australian patients compared to non-Indigenous Australian patients. The incidence of positive Extractable Nuclear Antigen (ENA) was also higher among Indigenous Australian patients (84.62% vs 57.14%). CONCLUSION Our study further supports the observation that lupus in Aboriginal and Torres Strait Islander Australians were of a 'distinct phenotype' compared to non-Indigenous Australians. Future research should be aimed at delineating the reason for this observed difference. This article is protected by copyright. All rights reserved.

Autoantibodies to glycine and tryptophan-rich bodies (GWB) can be detected on routine antinuclear antibodies (ANA) testing and might have important disease associations. The aim of this study was to investigate the prevalence of anti-GWB antibodies identified on routine ANA testing, define their antigenic specificities and describe their clinical association. Anti-GWB antibodies were identified by distinct cytoplasmic staining pattern on all samples referred for ANA testing over a 6-month period. All positive anti-GWB samples were further tested on a multiplex addressable bead immunoassay (ALBIA) with known GWB antigens. Extractable nuclear antigens (ENA) were characterised by line immunoblot assay. Clinical details were collected retrospectively by contacting patients and the requesting clinicians. Eleven patients (7 females, 4 males) out of a total of 2136 positive ANAs requested on 11,265 samples had the classical GWB pattern (0.5%). The median age of patients was 66 years (range 39-92). There was no consistent disease association. Ten were confirmed to have distinct antigenic specificity for known GWB antigens. Ge-1/Hedls and RAP55 were the most common antigenic specificity targets [seen in 7 patients (64%) and in 5 patients (45%), respectively]. Ro52 was positive in 5/9 (56%) patients, SSB in 2/9 (22%) patients and Ro60 in 1/9 (11%) patient. The clinical association of anti-GWB antibodies is uncertain but might point towards autoimmune origin of certain non-specific musculoskeletal symptoms. The antigenic specificity of anti-GWB reactivity could point towards specific clinical associations: anti-RAP55 and Ge-1 in non-specific musculoskeletal conditions versus anti-GW182 in neurological diseases.

Purpose Testing for autoantibodies to extractable nuclear antigens (ENAs) plays an important role in the diagnosis and management of systemic rheumatic disease. Currently, no gold standard tests are available for detecting anti-ENAs. To address this gap, we aimed to identify an assay that exhibits satisfactory diagnostic performance in the detection of five common anti-ENAs by comparing two commonly used assays, an automated fluorescent enzyme immunoassay (FEIA) and a microplate ELISA assay. Materials and methods Sera from 100 patients with systemic rheumatic disease were collected and assayed with FEIA and microplate ELISA to detect anti-ENAs. Statistical analyses were performed to check the agreement rate between the two platforms using kappa coefficients. Analytical sensitivity and specificity for each assay were calculated. Results The concordance rates between ELISA and FEIA ranged from 89% for anti-RNP to 97% for anti-Scl-70, and the kappa coefficients of the two assays were in the range of 0.44 to 0.82. Between the two assays, a significant difference in sensitivity and specificity was seen only for anti-Sm and anti-RNP, respectively. Conclusion In this study, FEIA and ELISA showed comparable efficiency for detecting anti-ENAs.

The role of autoantibody testing for patients with interstitial lung disease is an evolving area. Recent guidelines recommend routine anti-nuclear antibodies, rheumatoid factor, and anti-citrullinated cyclic peptide antibody testing for patients undergoing diagnostic evaluation for interstitial lung disease, with further autoantibody testing reserved for selected cases guided by rheumatological features. Even this approach may miss patients with clinically significant autoantibodies when interstitial lung disease is the dominant or first manifestation of autoimmune disease. We retrospectively performed autoimmune serology in a clinically well characterised cohort of interstitial lung disease patients. Using stored serum, additional testing was performed to ensure all patients had complete autoantibody profiles including anti-nuclear antibodies, extractable nuclear antigen antibodies, double-stranded DNA antibodies, rheumatoid factor, anti-citrullinated cyclic peptide antibodies, anti-neutrophil cytoplasmic antibodies, and myositis antibodies. Eighty patients with interstitial lung disease, and available stored serum, were assessed. Mean age at interstitial lung disease diagnosis was 65.2 years and 42 patients were male. Positive autoimmune serology was found in 56 of 80 (70.0%) patients; the most common positive result was anti-nuclear antibodies (n=34; 42.5%). Myositis antibodies were detected in 13 of 80 (16.2%) patients. Four (5%) patients had elevated anti-citrullinated cyclic peptide antibodies, and two (2.5%) patients had detectable myeloperoxidase antibodies. Eleven (13.7%) patients with negative anti-nuclear antibodies had other significant disease associated autoantibodies. An extended panel of autoantibody testing may detect cases of connective tissue disease associated interstitial lung disease, regardless of clinical or radiological subtype, and prior to extra-pulmonary manifestations of systemic autoimmunity.

Background The importance of systemic sclerosis (SSc) autoantibodies for diagnosis has become recognized by their incorporation into the 2013 ACR/EULAR classification criteria. Clear prognostic and phenotypic associations with cutaneous subtype and internal organ involvement have been also described. However, little is known about the potential of autoantibodies to exert a direct pathogenic role in SSc. The aim of the study is to assess the pathogenic capacity of anti-DNA-topoisomerase I (anti-Topo-I) and anti-centromeric protein B (anti-Cenp-B) autoantibodies to induce pro-fibrotic markers in dermal fibroblasts. Methods Dermal fibroblasts were isolated from unaffected and affected skin samples of (n = 10) limited cutaneous SSc (LcSSc) patients, from affected skin samples of diffuse cutaneous (DcSSc) patients (n = 10) and from healthy subjects (n = 20). Fibroblasts were stimulated with anti-Topo-I, anti-Cenp-B IgGs, and control IgGs in ratios 1:100 and 1:200 for 24 h. Cells were also incubated with 10% SSc anti-Topo-I+ and anti-Cenp-B+ whole serum and with 10% control serum for 24 h. Viability was assessed by MTT test, while apoptosis was assessed by flow cytometry. Activation of pro-fibrotic genes ACTA2, COL1A1, and TAGLN was evaluated by quantitative real-time PCR (qPCR), while the respective protein levels alpha-smooth-muscle actin (α-SMA), type-I-collagen (Col-I), and transgelin (SM22) were assessed by immunocytochemistry (ICC). Results MTT showed that anti-Cenp-B/anti-Topo-I IgGs and anti-Cenp-B+/anti-Topo-I+ sera reduced viability (in a dilution-dependent manner for IgGs) for all the fibroblast populations. Apoptosis is induced in unaffected LcSSc and control fibroblasts, while affected LcSSc/DcSSc fibroblasts showed apoptosis resistance. Basal mRNA (ACTA2, COL1A1, and TAGLN) and protein (α-SMA, Col-1, and SM22) levels were higher in affected LcSSc/DcSSc fibroblasts compared to LcSSc unaffected and to control ones. Stimulation with anti-Cenp-B/anti-Topo-I IgGs and with anti-Cenp-B+/anti-Topo-I+ sera showed a better induction in unaffected LcSSc and control fibroblasts. However, a statistically significant increase of all pro-fibrotic markers is reported also in affected LcSSc/DcSSc fibroblasts upon stimulation with both IgGs and sera. Conclusions This study suggests a pathogenic role of SSc-specific autoantibodies to directly induce pro-fibrotic activation in human dermal fibroblasts. Therefore, besides the diagnostic and prognostic use of those autoantibodies, these data might further justify the importance of immunosuppressive drugs in the early stages of the autoimmune disease, including SSc. Electronic supplementary material The online version of this article (10.1186/s13075-019-1931-x) contains supplementary material, which is available to authorized users.

Objective: The aim of this study was to investigate the predictive value of positive enzyme immunoassay (EIA) for detection of specific autoantibodies in sera negative for antinuclear antibody (ANA) by indirect immunofluorescence (IIF) assay, but positive for ANA by EIA. Methods: Eighty sera that tested negative for ANA by IIF, but positive for ANA by EIA were included. All sera were tested for specific autoantibodies by line immunoassay (LIA). The positive predictive value (PPV) of EIA was calculated using LIA result as a reference standard. Medical records of patients were reviewed. Clinical findings at the time of blood sampling for ANA testing and at 5 years after sampling were obtained. Results: Twenty-eight sera (35%) were found to be positive by LIA. The PPV of EIA for detection of specific autoantibodies at the manufacturer’s recommended cut-off was 35.0% (95% CI: 24.5-45.5%). The most prevalent antibodies were anti-SSA/Ro60 (64.3%, 95% CI: 46.5-82.0%), anti-Ro52 (25.0%, 95% CI: 9.0-41.0%), and anti-SSB/La (10.7%, 95% CI: 0-22.2%). A diagnosis of systemic autoimmune rheumatic disease was established in 7 patients (25%) at the time of blood sampling, and 4 patients (14.3%) were diagnosed with non-rheumatic autoimmune disease. Conclusion: EIA testing in IIF-ANA negative sera yielded a chance to detect antinuclear antibodies. However, the poor PPV of EIA may have low benefit in real-life clinical practice. Anti-SSA/Ro60 was the most prevalent antibody detected. A high proportion of LIA-ANA positive patients were not diagnosed as autoimmune disease at the time of antibody detection.

BACKGROUND Extractable nuclear antigen (ENA) and anti-double stranded DNA (anti-dsDNA) positivity and related diseases like systemic lupus erythematosus, Sjogren syndrome, and other autoimmune diseases are known to be associated with obstetrical complications and poor perinatal outcomes. OBJECTIVE To demonstrate the importance of ENA, anti-dsDNA, antiphospholipid (APL), and anticardiolipin (ACL) antibody positivity on pregnancy outcomes. METHODS Ninety one pregnant women with known ENA, anti-dsDNA, APL IgG and IgM, and ACL IgG and IgM antibody positivity were retrospectively compared with 91 randomly selected pregnant woman in terms of obstetrical complications and pregnancy outcomes. Beksac Obstetrics Index-pregnancy (BOIp), calculated as (number of children + (π/10))/gravidity in the current pregnancy, was used to compare the risk level between groups. RESULTS Significant differences were found in the median maternal age, gravidity, number of previous miscarriages, and BOIp between the groups (p= 0.04, p< 0.001, p< 0.001, and p< 0.001, respectively). Significant differences were also found between the study and control groups in the median gestational age at birth, birth weight, and APGAR1 score (p< 0.001 for all). Similarly, significant differences were found between groups in the rates of intra-uterine growth restriction, oligohydramnios, and gestational hypertension (p< 0.001, p= 0.05, and p= 0.05, respectively). There were 3 (3.3%) stillbirths in the study group and none in the control group (p= 0.123). CONCLUSION We evaluated the impact of anti-dsDNA, ENA, APL, and ACL antibody positivity, which may cause immunologic inflammation at placenta and thereby affect pregnancy outcomes.

Background : Systemic Lupus Erythematosus (SLE) is said to be rare in Sub-Saharan Africa and even rarer in males worldwide. SLE is mostly considered a disease of women, though men may also be affected, and this may lead to a delay in diagnosis in men. The result is a greater burden of inflammation and subsequent organ damage over time. Method : Data from the medical records of 13 male patients diagnosed with SLE at the Rheumatology Clinic of Korle-Bu Teaching Hospital between January 2014 and January 2017 was retrospectively analyzed. Results : A total of 13 male patients out of a total of 134 SLE patients were included in our analysis. The mean age was 30.62 ± SD 8.47 years (range of 17 to 46 years). All of them (100%) presented with constitutional features. The most common ACR criteria observed was 61.5 % rash, 54.5 % oral ulcers, 92.3% arthritis, 61.5 % serositis and 38.5% renal involvement, 46.2 % CNS involvement. Looking at their serological profile, 91.7 % had a positive antinuclear antibody (ANA). 33.3 % had positive anti-dsDNA and 58.3 % extractable nuclear antigens. The mean duration from onset of symptoms to diagnosis was 21.31 months. Five patients were diagnosed with lupus nephritis, all at the time of diagnosis. There were no mortalities. Conclusion : Male SLE patients in Ghana are comparable to other populations, with arthritis and constitutional features being predominant early features and lupus nephritis being the main early indicator of organ damage. This should warrant aggressive management in male patients. Funding : None declared Keywords : Male, systemic lupus erythematosus, nephritis, Sub-Saharan Africa

As iconic and important diagnostic autoantibodies, anti-Ro60 and anti-Ro52/tri-partite motif-containing 21 (TRIM21) make a common appearance in a number of systemic autoimmune disorders such as systemic lupus erythematosus (SLE). These autoantibodies often co-exist together; yet despite their close relationship, there is no evidence that they are physically linked and probably reflect a convergence of separate processes of failed immunological tolerance. Confusingly, they are sometimes classed together as the "SSA" or "Ro" autoantibody system without clear distinction between the two. In this Short Communication, we discuss the diagnostic merits for separate detection and reporting of these two autoantibodies, and discuss avenues for future research. Indeed, further insight into their fascinating origins and pathogenic roles in autoimmunity will surely shed light on how we can prevent and treat devastating autoimmune disorders.

Objective LIA-ANA-Profile-17S is a multiplex line immunoassay that simultaneously detects 17 antinuclear antibodies (ANAs) against extractable nuclear antigens (ENAs). We evaluated the utility of LIA-ANA-Profile-17S as a supplement to ANA indirect immunofluorescence (IIF) and EliA ENA (a fluorescence enzyme immunoassay) for diagnosis of ANA-associated rheumatic diseases. Methods Sera were collected from 245 patients referred for an ANA IIF test. LIA-ANA-Profile-17S results were compared with those of EliA ENA. The kappa coefficients, agreement rates, and diagnostic performance of these tests were assessed for systemic lupus erythematosus (SLE) and Sjögren’s syndrome (SjS). Results We observed almost perfect interassay agreement for antibodies against Ro52/Ro60, CENP-B, and Scl-70 (kappa = 0.91, 0.97, and 1.00, respectively); strong agreement for anti-SS-B/La antibody (kappa = 0.81); and relatively low agreement for other antibodies, including those against dsDNA, Sm, RNP, and Jo-1. For SLE diagnosis, LIA-ANA-Profile-17S showed lower sensitivity and similar specificity compared with EliA ENA. The sensitivity and specificity of these two assays were similar for SjS diagnosis. Conclusions The specificity of LIA-ANA-Profile-17S was enhanced when combined with ANA IIF and was comparable with that of EliA ENA. LIA-ANA-Profile-17S showed relatively good agreement with EliA ENA. In combination with ANA IIF, these assays showed enhanced diagnostic performance.

To describe the clinical manifestations, immunological features, and risk factors in patients with sarcoidosis complicated with autoimmune diseases (ADs) as well as determine the frequency of autoantibodies and possible correlation between autoantibodies and laboratory data. Patients with pathologically confirmed sarcoidosis at Beijing Chaoyang Hospital (China) between January 2017 and October 2020 were included. Age- and sex-matched patients who visited the rheumatology outpatient clinic without systemic or ADs were included as controls. Demographic, clinical, serological, and radiological data of sarcoidosis patients were recorded and analyzed. To exclude ADs, autoantibodies, such as antinuclear antibody, extractable nuclear antigen antibodies, and anti-cyclic citrullinated peptide antibody were assessed in controls. A total of 154 sarcoidosis patients (111 females; 72.1%) with a mean ± standard deviation age of 50.7 ± 10.3 years were included. Nineteen patients (12.3%) had ADs; Hashimoto’s thyroiditis (n = 6) and Sjogren's syndrome (n = 4) were common. Age, globulin, immunoglobulin G, erythrocyte sedimentation rate (ESR), and C-reactive protein were significantly different between sarcoidosis patients with and without ADs. The ESR level might be a risk factor for sarcoidosis complicated with ADs (RR = 1.053; P = 0.018). Autoantibodies were detected in 29 patients (18.8%), and the frequency was significantly higher than that in controls (18.8% vs. 3%; P = 0.001). Sarcoidosis patients were more likely to have autoantibodies despite the absence of ADs (10.4% vs. 3%; P = 0.031). Age may be a risk factor for sarcoidosis patients presenting with autoantibodies (RR = 1.077; P = 0.042). An association was identified between ADs and sarcoidosis. The inflammatory indexes, such as ESR, IgG, and CRP, were significantly different between sarcoidosis patients with and without ADs. ESR might be a risk factor for the coexistence of ADs and sarcoidosis. Sarcoidosis patients were prone to being autoantibody-positive despite the absence of ADs, and age might be a risk factor for sarcoidosis presenting with autoantibodies.

Systemic Lupus Erythematosus (SLE) is a clinico-serologic diagnosis. The screening test for SLE is antinuclear antibody (ANA) by indirect immunofluorescence. A positive ANA does not confirm SLE. Clinical context is crucial in the interpretation of a positive test result. A negative ANA virtually rules out SLE. ANA patterns give an idea about the antigens targeted and the underlying autoimmune disease. The correlation at best is general and should not be used to guide treatment decisions. ANA is sensitive while antibodies to extractable nuclear antigens (ENA) are specific. ENA should not be used as a screening test. Changes in ANA titer do not parallel disease activity. Therefore, repeat testing is discouraged. Urinalysis is a must for all patients at all visits. Double-stranded deoxyribonucleic acid (dsDNA) antibodies, complement levels, and urinalysis are used to monitor the activity of lupus nephritis. Treatment in clinically quiescent patients should not be escalated based only on persistent serological activity.

Duran, Ali (Balikesir Author), Autoantibodies targeting nuclear and cytoplasmic autoantigens are used as markers in the diagnosis and classification of systemic autoimmune rheumatic diseases (SARD). The dense fine speckled (DFS) pattern is characterized by the fine-granular fluorescence of the nuclei in the interphase and the metaphase chromatin. DFS70 antibodies have been reported in healthy individuals, various autoimmune disorders, infection, cancer and inflammatory conditions. But there is still lack of information about its clinical significance. This study aimed to investigate the clinical significance of anti-DFS70 autoantibodies and the determination of accompanying pathologies. A total of 5710 serum samples routinely requested for ANA screening were tested between 2017 and 2019. Antinuclear antibody (ANA) and dsDNA were performed by indirect immunofluorescence method (IIF) (Euroimmun, Germany). Immunoblot (IB) method was used for the extractable nuclear antigen profile (ENA) (Euroimmun, Germany). Demographic and clinical data, were investigated from the medical records. Among 5710 samples tested for ANA, 23.7% were ANA positive by IIF. Mean age of the patients were 47.9 and 79.5% were female. Only 8.1% of the study group had SARD. The frequency of DFS pattern by ANA-IIF was 6.0% (342/5710), (mean age ± SD= 44.4 ± 16.7, 88% female). DFS70 pattern-positive patients were sub-grouped according to their diagnosis. SARD were detected 10.8% (mean age ± SD= 55.12 ± 14.10) in DFS70 pattern positive patient group (RA 6.1%, SS 2.6%, SLE 0.9%, SSc 0.6%, UCTD 0.6%). Autoantibodies accompanying anti-DFS70 antibody were determined as Ro-52, SS-A, nucleosome, histone, AMA-M2, dsDNA, respectively. Non-SARD diseases were determined in 89.2% of the patients with positive DFS70 pattern. Non-SARD diseases were detected as musculoskeletal complaints (47.4%), other rheumatic diseases like fibromyalgia (14.3%), dermatological diseases (9.4%), gastrointestinal system diseases (5.6%), hematological disorders (3.8%), thyroid /parathyroid diseases (3.5%), allergic diseases (2.3%), neurological diseases (2.3%) and neoplasia (breast cancer) (0.6%). The anti-DFS70 autoantibody is widely used to exclude the diagnosis of SARD in the absence of concomitant SARD-related autoantibodies. It has been observed that anti-DFS70 autoantibody may be associated with non-SARD rheumatic diseases and in many diseases (dermatological, gastrointestinal system, hematological, thyroid diseases) related to other systems. Therefore it is essential to evaluate these pathologies in patients positive for anti-DFS70 antibodies., Nükleer ve sitoplazmik otoantijenleri hedefleyen otoantikorlar, sistemik otoimmün romatizmal hastalıkların (SORH) tanı ve sınıflandırılmasında belirteç olarak kullanılmaktadır. Dense fine speckled (DFS) paterni interfaz evresinde nükleusda ve metafaz kromatininde yoğun ince benekli tarzda boyanma gösterir. Anti-DFS70 antikorları sağlıklı kişilerde, çeşitli otoimmün bozukluklarda, enfeksiyonda, kanserde ve enflamatuvar durumlarda bildirilmiş olmakla birlikte klinik önemi hakkında bilgiler hala yetersizdir. Bu çalışmada, anti-DFS70 otoantikorlarının klinik öneminin araştırılması ve eşlik eden patolojilerin belirlenmesi amaçlanmıştır. 2017-2019 yılları arasında rutin ANA taraması için gönderilen toplam 5710 serum örneği test edilmiştir. Örnekler antinükleer antikor (ANA) ve dsDNA indirekt immünfloresan yöntemleri (IIF) (Euroimmun, Almanya) ile çalışılmıştır. Ekstrakte edilebilir nükleer antijen (ENA) profili için immünoblot (IB) yöntemi (Euroimmun, Almanya) kullanılmıştır. Demografik ve klinik veriler, tıbbi kayıtlar incelenerek elde edilmiştir. ANA-IIF ile test edilen 5710 örneğin %23.7’si ANA pozitif bulunmuştur. Hastaların ortalama yaşı 47.9 ve %79.5’i kadın olarak tespit edilmiş ve çalışma grubunun sadece %8.1’inde SORH saptanmıştır. IIF-ANA testine göre DFS70 paterni sıklığı %6.0 (342/5710) olarak belirlenmiştir (Ortalama yaş ± Ss= 44.4 ± 16.7,%88 kadın). DFS70 paterni saptanan hastalar tanılarına göre alt gruplara ayrılmıştır. DFS70 paterni pozitif hasta grubunda %10.8 oranında (ortalama yaş ± Ss= 55.12 ± 14.10) SORH saptanmıştır (RA %6.1, SS %2.6, SLE %0.9, SSc %0.6, UCTD %0.6). Anti-DFS70 antikoruna eşlik eden otoantikorlar sırasıyla Ro-52, SS-A, nükleozom, histon, AMA-M2, dsDNA olarak belirlenmiştir. Anti-DFS70 paterni pozitif hastaların %89.2’sinde SORH dışı hastalıklar saptanmıştır. Kas iskelet sistemi şikayetleri (%47.4), fibromiyalji gibi diğer romatizmal hastalıklar (%14.3), dermatolojik hastalıklar (%9.4), gastrointestinal sistem hastalıkları (%5.6), hematolojik bozukluklar (%3.8), tiroid/paratiroid hastalıkları (%3.5), alerjik hastalıklar (%2.3), nörolojik hastalıklar (%2.3) ve neoplazi (meme kanseri) (%0.6) SORH dışı hastalıklar olarak belirlenmiştir. Anti-DFS70 otoantikoru, eşlik eden SORH ile ilişkili otoantikorların yokluğunda SORH teşhisini dışlamak için yaygın olarak kullanılmaktadır. Anti-DFS70 otoantikorunun SORH dışı romatizmal hastalıklarla ve diğer sistemlerle ilgili birçok hastalıkla (dermatolojik, gastrointestinal sistem, hematolojik, tiroid hastalıkları) ilişkili olabileceği gözlenmiştir. Bu nedenle, anti-DFS70 antikorları pozitif hastalarda bu patolojilerin değerlendirilmesi önemlidir.

Rheumatology is a clinically oriented specialty. All laboratory tests need to be interpreted in light of the clinical picture since tests, viewed in isolation, can be misleading. Rheumatoid Factor (RF) is seen in several conditions, including Rheumatoid Arthritis (RA). A positive RF does not mean RA. A negative test does not exclude RA. Indirect immunofluorescence (IIF) is the preferred test methodology for Antinuclear antibodies (ANA). A negative ANA on IIF virtually rules out systemic lupus erythematosus (SLE) because ANA-negative SLE is very rare in clinical practice. A positive test may or may not confirm SLE and needs clinical correlation. Antibodies to extractable nuclear antigen (ENA) should only be obtained in patients with positive ANA. Antiphospholipid syndrome is a clinico-serologic diagnosis that requires clinical features in addition to antiphospholipid antibodies. Antistreptolysin O should not be used as the only basis for diagnosing rheumatic fever. ELISA is the preferred test methodology for Antineutrophil cytoplasmic antibody (ANCA) detection. ANCA should not be used as a screening test. A high serum uric acid does not translate into a diagnosis of gout. Hyperuricemia is a biochemical abnormality while gout is a clinical syndrome. Synovial fluid examination is an important yet infrequently ordered test. It is invaluable in monoarthritis. HLA B27 is present in a significant proportion of the healthy population, the majority of whom will never develop SpA.

Background The International Consensus on Antinuclear Antibody (ANA) Patterns (ICAP) has recently proposed nomenclature in order to harmonize ANA indirect immunofluorescence (IIF) pattern reporting. ICAP distinguishes competent-level from expert-level patterns. A survey was organized to evaluate reporting, familiarity, and considered clinical value of ANA IIF patterns. Methods Two surveys were distributed by European Autoimmunity Standardization Initiative (EASI) working groups, the International Consensus on ANA Patterns (ICAP) and UK NEQAS to laboratory professionals and clinicians. Results 438 laboratory professionals and 248 clinicians from 67 countries responded. Except for dense fine speckled (DFS), the nuclear competent patterns were reported by > 85% of the laboratories. Except for rods and rings, the cytoplasmic competent patterns were reported by > 72% of laboratories. Cytoplasmic IIF staining was considered ANA positive by 55% of clinicians and 62% of laboratory professionals, with geographical and expertise-related differences. Quantification of fluorescence intensity was considered clinically relevant for nuclear patterns, but less so for cytoplasmic and mitotic patterns. Combining IIF with specific extractable nuclear antigens (ENA)/dsDNA antibody testing was considered most informative. Of the nuclear competent patterns, the centromere and homogeneous pattern obtained the highest scores for clinical relevance and the DFS pattern the lowest. Of the cytoplasmic patterns, the reticular/mitochondria-like pattern obtained the highest scores for clinical relevance and the polar/Golgi-like and rods and rings patterns the lowest. Conclusion This survey confirms that the major nuclear and cytoplasmic ANA IIF patterns are considered clinically important. There is no unanimity on classifying DFS, rods and rings and polar/Golgi-like as a competent pattern and on reporting cytoplasmic patterns as ANA IIF positive.

Background Neuromyelitis optica spectrum disorders (NMOSD) are considered as an autoantibody-mediated disorder that targets aquaporin-4 (AQP4); other autoantibodies could be detected in such spectrum of diseases, including anti-nuclear antibody and antibodies to extractable nuclear antigens. Systemic autoimmune diseases such as systemic lupus erythematosus (SLE), Sjogren’s syndrome (SS), and other autoimmune diseases can overlap with NMOSD. We aimed in this review to address the current evidence describing the relation of NMOSD to systemic autoimmunity diseases, its controversy of being co-association or the same etiology, and its practical implications. Main body The current review was done using a search for related articles or case reports on PubMed until 2019. The keywords included neuromyelitis optica spectrum disorders in combination with autoimmune disease nomenclature. We described the literature background of this controversy, to summarize the evidence of NMOSD relationship to systemic autoimmune diseases. Conclusion NMOSD associated with systemic autoimmune diseases is more common in SLE and Sjogren’s syndrome rather than other autoimmune diseases, frequently affects females more than males; AQP4 antibodies should be tested for all NMOSD like manifestations associated with an autoimmune disorder; however, the clinical diagnosis of NMOSD regardless of the cord lesion length and the presence of positive AQP4 antibody can occur in systemic autoimmune diseases.

Detection of autoantibody against extractable nuclear antigens (ENAs) plays a critical role in the diagnosis and management of autoimmune diseases.In this study, we assessed the performance of LETIA in detecting anti-ENAs. Total 606 serum samples from the Second Hospital of Shanxi Medical University were collected. Anti-SSA, anti-SSB, anti-Sm, anti-U1-snRNP, and anti-Sm/RNP were parallelly detected by LETIA and line immunoblot (LIA). Besides, this study assessed LETIA for its repeatability in detecting anti-ENAs autoantibodies, and consistency with LIA. A receiver operating characteristic (ROC) curves was drawn to assess the accuracy of LETIA. The LETIA and LIA showed high coincidence rate in detecting anti-SSA, anti-SSB, anti-Sm, anti-U1-snRNP, and anti-Sm/RNP autoantibodies, with the results being 87.22%, 96.61%, 97.03%, 88.28%, and 92.06%, respectively. Almost perfect consistency (kappa > 0.8) were found in the detection of anti-SSB and anti-Sm by LETIA and LIA. While in the detection of anti-SSA, anti-U1-snRNP, and anti-Sm/RNP, moderate consistency (0.6 ⩽ kappa ⩽ 0.8) were shown. The AUCs of anti-SSA, anti-SSB, anti-Sm, anti-U1-snRNP, and anti-Sm/RNP detected by LETIA were 0.972 (95% confidence interval (CI): 0.941–1.000, p < 0.001), 0.986 (95% CI: 0.967–1.000, p < 0.001), 0.912 (95% CI: 0.763–1.000, p < 0.001), 0.829 (95% CI: 0.731–0.928, p < 0.001), and 0.828 (95% CI: 0.715–0.941, p < 0.001), respectively. LETIA and LIA showed considerable consistency in detecting anti-ENAs. Moreover, with the pronounced advantages of automatic and rapid detection, and high universality, LETIA can meet the requirements for quantitative detection of anti-ENAs.

International audience; PurposeThe goal of this study is to determine a link between benign essential blepharospasm and Sjogren's syndrome by analyzing the presence of extractable nuclear antigens in this population.MethodsSeventy-two patients with benign essential blepharospasm (BEB) were included in this study. We eliminated patients with hemifacial spasm or blepharospasm secondary to corneal pathology. We collected the values of the Schirmer I test and the results of the anti-SSA and anti-SSB antibodies.ResultsOur study included 72 patients (144 eyes) whose 62 women (86.1%). Mean age was 74.3 years ± 10.73. Average Schirmer I test was 3.14 mm ± 4.00 mm. Five women (8% of this female population) had positive anti-SSA and SSB antibodies. Their mean age was 65.66 years ± 13.24 whereas the negative antibody patients had an average age of 75.42 ± 9.27. There was no significant difference between their Schimer I test and the Schirmer I of negative antibody population.ConclusionThis study illustrates the possible association between the presence of Sjögren's syndrome and the occurrence of a BEB justifying the search for anti-SSA and anti SSB in blepharospasm patients.; ObjectifÉtudier s’il existe un lien entre le blépharospasme essentiel et le syndrome de Sjögren en analysant la présence d’anticorps nucléaires solubles dans cette population.MéthodesSoixante douze patients atteints de blépharospasme essentiel (BSE) ont été inclus dans cette étude. Nous avons éliminé les patients atteints de spasme hémifacial, ou de blépharospasme secondaire à une pathologie cornéenne avérée. Nous avons recueilli les valeurs des test de Schirmer I et les résultats des anticorps anti-SSA et anti-SSB.RésultatsNotre étude a regroupé 72 patients (144 yeux) dont 62 femmes (86,1 %). L’âge moyen était de 74,3 ans ± 10,73. Le test de Schirmer I était en moyenne de 3,14 mm ± 4,00 mm. Cinq femmes (8 % de cette population féminine) ont révélés des anticorps anti-SSA et SSB positifs. Leur moyenne d’âge était de 65,66 ans ± 13,24 alors que les patients anticorps négatifs avaient une moyenne d’âge de 75,42 ± 9,27. Il n’existait pas de différence statistiquement significative entre leur test de Schimer I et celui des patients anticorps négatifs.ConclusionCette étude illustre la possible association entre la présence d’une maladie de Sjögren et la présence d’un BSE justifiant la recherche d’anticorps anti-SSA et SSB chez ces patients.

INTRODUCTIONS To analyse the clinical significance of anti-Ro52 autoantibodies that target TRIM21 protein and occur in a variety of connective tissue diseases (CTD), 4782 cases with blood tested positive for anti-Ro52 antibodies between January 2016 and September 2017 in Drum Tower Hospital were enrolled. METHODS Anti-Ro52 and anti-Ro60 were measured using the immunoblotting method together with antibodies against other extractable nuclear antigens at the time of the patient's first visit. RESULTS In this cohort, females accounted for the majority of all subjects (75.9%) and the average age was 47.25 years old. About 97.4% were diagnosed as having various diseases, in which around 2/3 were CTDs. The incidence of CTD in patients with merely anti-Ro52 was lower than those with both anti-Ro52 and anti-Ro60 (54.7% vs 85.5% for all patients and 53.3% vs 87.1% for inpatients, both P

Background:Some infectious agents may act as inducers of autoimmune conditions1. Despite SARS-CoV-2 infection can induce autoimmune phenomena in infected people2, individual risk factors or underlining mechanisms leading to loss of immunological tolerance are still unknown.Objectives:To assess the rate of development of autoantibodies in convalescent SARS-CoV-2 patients and their relation on infection clinical course and disease parameters.Methods:One-hundred and nine convalescent SARS-CoV-2 patients were studied and underwent multidisciplinary assessment in a Day Hospital clinical setting. For each patient, demographic, clinical and immunological data were collected and, at study entry, autoimmune profile [antinuclear antibodies (ANAs), antibodies reacting with extractable nuclear antigens (anti-ENA), antineutrophil cytoplasmic antibodies (ANCAs), Lupus anticoagulant (LA), anticardiolipin antibodies (aCL) IgM and IgG, anti-β2 glycoprotein I (anti-β2GPI) IgM and IgG] was assessed by Fluorescent immunoassay. Moreover, IL-6 plasma levels were assessed by ELISA (ELLA).Results:After a median time from hospitalization for SARS-CoV-2 infection of 53.3 ± 0.9 days, 55(50.5%) SARS-CoV-2 convalescent patients showed the positivity (ABpos) of at least one autoantibody. In particular, 31(28.4%) were positive for LA, 11(10.1%) for IgM-RF, 8(7.3%) for ANA and 6(5.5.%) for IgG-aCL whilst less than 2% showed other autoantibody positivities (IgM-aCL, IgG-anti-β2GPI, ENA, ACPA, c-ANCA, Scl70 and RNP). Analyzing the patient-related characteristics associated with the development of autoimmunity, convalescent male patients were more likely characterized by the development of antiphospholipid antibodies (aPL) (37.3%) than female (16.7%; p=0.02). Considering the disease-related characteristics, convalescent SARS-CoV-2 patients who experienced severe pneumonia (i.e., oxygen support need) during hospitalization, more likely received IL-6R-inhibitor administration (47.3%) and developed more than one autoantibody (87.5%) (aPL + another AB) than convalescent SARS-CoV-2 patients who did not need oxygen support [(12.5%; p=0.02) (OR95%IC: 9.5(1.4-109.1)] or IL-6R-inihibitor (1.9%; pnegpatients (5.6 ± 1.5 pg/ml; p=0.07), mostly if considered autoantibodies other than aPL (15.4 ± 7.7 pg/ml)(p=0.01).Conclusion:Loss of self-tolerance is a common phenomenon in the medium-term follow-up of SARS-CoV-2 convalescent patients whose occurrence is dependent by a severe disease course and by an aberrant host inflammatory response. Long-term follow-up will reveal AB persistency and their clinical impact.References:[1]Barzilai O, et al. Current Opinion in Rheumatology 2007.[2]Zhou Y, et al. Clin Transl Sci. 2020.Disclosure of Interests:None declared

Background/Purpose Anti-carbamylated protein antibodies (anti-CarP) have been described not only in Rheumatoid arthritis but in other systemic autoimmune diseases. Recently, they have been reported in different cohorts of Systemic Lupus Erythematosus (SLE) with a prevalence of 9–28%1–4 in patients selected with arthritis/arthralgias. Anti-CarP have been proposed as a marker of erosive arthritis in SLE.4 The aim was to assess the prevalence of anti-CarP in SLE patients from a single center cohort and their association to clinical and laboratory data. Methods Serum anti-CarP levels were evaluated using a home-made ELISA (nv Results Complete clinical and serological data were available for 314 consecutive patients: 85 (27%) positive and 229 (73%) negative. No association was found among CarP+ and arthritis/arthralgias. CarP+ patients presented an earlier disease onset compared with CarP- (mean 28±11vs32±14.7,p=0.001), a trend towards a higher prevalence of xerophtalmia (36% vs 26.5%, p=0.075;OR:1.62,95%CI:0.95–2.75) and extractable nuclear antigen positivity (67% vs 54%,p=0.064;OR:1.65,95%CI:0.97–2.8). Interestingly, patients anti-CarP+ less frequently experienced class IV glomerulonephritis (12%vs 21.8%,p=0.05;OR:0.53,95%CI:0.26–1.08). Fifty-six patients evaluated were treated with anti-Blys therapy and longitudinally sera were available (T0,T6,T12). At baseline anti-CarP were positive in 10 (17.8%) and titre significantly decreased at T6 (p=0.006) and T12 (p=0.01). Negative seroconversion was observed in 7/10 sera. Conclusions The prevalence of anti-CarP antibodies found in our unselected cohort is in line with to what previously reported. In our hands, anti-CarP antibodies seems to identify a less severe form of SLE, with less kidney involvement and probably in overlap with Sjogren disease. Further studies are needed in order to be able to identify a possible role for this autoantibody. References Ziegelasch M,et al.Arthritis Res and Ther 2016;18:289. Nakabo S,et al J Rheumatol 2017 Pecani A,et al. Arthritis Res Ther 2016; 18: 276. Ceccarelli F,et al. Arthritis Res Ther 2018; 20:126.

Background This systematic review of literature and meta-analysis aimed to determine the prevalence, phenotype and treatment of systemic lupus erythematosus (SLE) in Native sub-Saharan Africans. Methods PubMed, EMBASE, Web of Science, African Journals Online, and Global Index Medicus as well as references of retrieved papers were searched to select studies addressing SLE in Native sub-Saharan Africans and published during January 1, 2008- October 7, 2018. Results were pooled through narrative review and random-effects model. Heterogeneity (I2) was assessed via the χ2 test. Pooled estimates are expressed with 95% confidence intervals. This study is registered with PROSPERO: registration number CRD42019139226. Results Fifteen hospital-based studies were included out of 1502 records. The pooled prevalence of SLE was 1.7% (0.8–2.9). The mean age at diagnosis ranged from 28.8 to 39.2 years. The female proportion was 88%-100%. Rheumatological (5.1%-99.9%), dermatological (4.3%-100%) and hematological (1.4–86.9%) manifestations were the commonest. Patients had a high seroprevalence for anti-ribonucleoprotein 57.9% (36.4–77.9), anti-Smith 53.5% (40.4–66.2), anti-Sjogren syndrome antigen A 45.6% (19.2–73.4) and anti-Sjogren syndrome antigen B 33.7% (13.6–57.6) autoantibodies. The most used treatments were corticosteroids 99% (94.9–100) and antimalarials 62. 8% (23.3–94.1). The pooled mortality rate was 10.3% (3.3–20.6); mainly due to infections, kidney and neurological involvement. Conclusions Over the last 30 years, SLE was not rare among Native sub-Saharan Africans and its featured characteristics were earlier onset, female predominance, and high seropositivity for extractable nuclear antigen autoantibodies. The standard treatments were corticosteroids and antimalarials. The mortality rate was high. Population prevalence and incidence as well as full description of SLE characteristics in Native sub-Saharan Africans are needed.

Objective: The aim of this retrospective study was to investigate the relationship between serum systemic autoantibodies and anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis. Methods: Thirty-nine patients with anti-NMDAR encephalitis were examined for serum systemic autoantibodies (antinuclear antibodies, extractable nuclear antigen autoantibodies, rheumatoid factors, and anti-neutrophil cytoplasmic antibodies), in comparison with 39 neuromyelitis optica spectrum disorder (NMOSD) and 78 healthy controls. Clinical features, cerebrospinal fluid characteristics, and outcomes were compared between the two subgroups of anti-NMDAR patients with positive and negative systemic autoantibodies, respectively. Results: Anti-NMDAR encephalitis patients had higher frequency of positive serum systemic autoantibodies than healthy controls (23.1 vs. 2.6%, p = 0.001) and lower frequency than NMOSD (23.1 vs. 48.7%, p = 0.018). No patients were diagnosed comorbidities with non-organ-specific autoimmune diseases. Consciousness disturbance was more frequent in autoantibodies positive group than in the negative group (88.9 vs. 40.0%, p = 0.02). Autoantibody positive group had a poorer outcome than autoantibody negative group (55.6 vs. 86.7%, p = 0.043). There was a negative correlation between serum autoantibodies and outcomes in anti-NMDAR encephalitis patients (r = −0.325, p = 0.044). Conclusion: Our data demonstrated serum systemic autoantibodies were more frequent in anti-NMDAR encephalitis patients than in healthy controls and less frequent than NMOSD, which were associated with higher severity of disease.

Background Detection of antibodies to extractable nuclear antigens (ENAs) is needed for the diagnosis in systemic autoimmune diseases. In this study, we compared three reagents using line immunoblot assay (LIA) or multiplex bead immunoassay for detecting the anti-ENAs. Methods A total of 89 sera were tested by 3 different assays: EUROASSAY Anti-ENA Profile (Euroimmune, Germany), Polycheck Autoimmune Test (Biocheck GmbH, Germany), and FIDIS™ Connective Profile (Biomedical Diagnostics, France). The following individual ENAs were investigated: Sm, SS-A (Ro), SS-B (La), Scl-70, Jo-1 and RNP. We reviewed medical records to investigate the discrepant results among three methods. Results Overall percent agreements were 96.1% between EUROASSAY Anti-ENA Profile and FIDIS™ Connective profile; 90.4% between EUROASSAY Anti-ENA Profile and Polycheck Autoimmune Test using the manufacturers’ cutoff; 96.4% between EUROASSAY Anti-ENA Profile and Polycheck Autoimmune Test using a upward cutoff; 90.4% between FIDIS™ Connective profile and Polycheck Autoimmune Test the manufacturers’ cutoff; and 96.4% between FIDIS™ Connective profile and Polycheck Autoimmune Test a upward cutoff. Conclusions The three assays showed excellent agreement with each other. With appropriate cutoff, the all three assays for six of the anti-ENA tests investigated in this study can be used in clinical laboratories for detecting the anti-ENAs.

Background/objective The criterion standard for anti-topoisomerase I antibody (anti-topo I antibody) testing in systemic sclerosis (SSc) uses immunodiffusion (ID) techniques, but enzyme-linked immunosorbent assay (ELISA) and multiple-bead technology are often used in current settings to save time and cost. Our aim was to assess the performance of the multiple-bead assay, ELISA, and ID testing methods. Methods We conducted a retrospective study of patients at the University of Michigan whose extractable nuclear antigen 10 autoantibody panel tested positive for the anti-topo I antibody by multiple-bead technology during a 1-year period. All samples positive by multiple-bead assay were sent to the RDL Laboratories and reflexed for ELISA, and all anti-topo I antibodies positive by ELISA were further tested by ID. Clinical data were reviewed by a rheumatologist and assessed for presence of SSc. Data were analyzed via frequency tables. Results Approximately 9500 extractable nuclear antigen 10 panels were ordered by physicians at the University of Michigan. Of these, 129 patients were positive for the anti-topo I antibody by multiple-bead assay, 51 were positive by multiple-bead assay and ELISA, and 21 were positive by multiple-bead assay, ELISA, and ID. We found that 26.4% of patients positive by multiple-bead assay, 47.1% positive by multiple-bead assay and ELISA, and 95.2% positive by multiple-bead assay, ELISA, and ID had SSc. Conclusions Multiple-bead assays have a high rate of false-positive results for the anti-topo I antibody in patients without clinical evidence of SSc. A stepwise approach of confirmation of positive multiple-bead assay results using both ELISA and ID improves the predictive value of antibody testing for the diagnosis of SSc.

1. Richard K. Vehe, MD* 2. Mona M. Riskalla, MD* 1. *Division of Pediatric Rheumatology, Department of Pediatrics, University of Minnesota and the University of Minnesota Masonic Children’s Hospital, Minneapolis, MN * Abbreviations: ANA: : antinuclear antibody CTD: : connective tissue disease CVD: : collagen vascular disease dsDNA: : doubled-stranded DNA DM: : dermatomyositis ENA: : extractable nuclear antigen JIA: : juvenile idiopathic arthritis MCTD: : mixed connective tissue disease NFC: : nail fold capillary RA: : rheumatoid arthritis RF: : rheumatoid factor RP: : Raynaud phenomenon RNP: : ribonucleoprotein SLE: : systemic lupus erythematosus Timely and accurate recognition of collagen vascular disorders (CVDs), and implementation of effective screening and referral processes for patients suspected of having a CVD, remain a challenge for many physicians. The result, too often, is unnecessary testing and referrals, and in some cases unnecessary anxiety for physicians, patients, and parents. After completing this article, readers should be able to: 1. Recognize the common clinical symptoms and signs of systemic lupus erythematosus, dermatomyositis, and scleroderma, and their distinction from common infectious mimics. 2. Recognize the testing that can clarify the likelihood of whether a child has a rheumatic disease, including the limited utility of early serologic testing for autoantibodies. 3. Recognize the prognosis and management objectives for these often-chronic disorders, and practical steps to ensure excellent outcomes. Timely and accurate recognition of collagen vascular diseases (CVDs), and implementation of effective screening and referral processes for patients suspected of having a CVD, remain challenging for many physicians. The result, too often, is unnecessary testing with questionable results, leading in some cases to unnecessary referrals and anxiety for physicians, patients, and parents. (1) Sometimes, CVDs are also referred to as connective tissue diseases (CTDs), but neither name fully captures their essential nature. These are not primarily heritable disorders affecting collagen-containing structures, or disorders of vascular development, but rather are immune-mediated inflammatory diseases. The immunologic targets vary, but the inflammatory disease often affects collagen-containing tissues, including the musculoskeletal system and many other tissues. The targeting of blood vessels can lead to manifestations ranging from reversible vasospasm of Raynaud phenomenon (RP) to vascular injury of 1 or more organ systems, often with serious consequences. The diseases that …

The aim was to explore possible correlations of antibodies to extractable nuclear antigens (ENA) with clinical manifestations and disease activity indices in systemic lupus erythematosus (SLE) patients. A total of 70 consecutive SLE patients (64 females) were included. Disease activity was assessed by SLE activity index (SLEDAI), and British Isles Lupus Assessment Group (BILAG). Anti-Ro/SSA correlated positively with, headache (r=0.24, p=0.04), blurring of vision (r=0.25, p=0.03) and SLEDAI (r=0.25, p=0.04) and negatively with C3 (r=–0.35, p=0.003). Anti-Ro/SSA correlated with anti La/SSB antibodies (r=0.69, p