scholarly journals Detection by Flow Cytometry of Anti-DNA Autoantibodies and Circulating DNA Immune Complexes in Lupus Erythematosus

2019 ◽  
Vol 2019 ◽  
pp. 1-12
Author(s):  
Nisen Abuaf ◽  
Chantal Desgruelles ◽  
Mohamed Moumaris ◽  
Faïza Boussa-Khettab ◽  
Hidayeth Rostane ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Lupus Erythematosus ◽  
Immune Complexes ◽  
Serum Samples ◽  
Crithidia Luciliae ◽  
Two Samples ◽  
Dna Antibodies ◽  
Diagnostic Sensitivity And Specificity ◽  
Polyethylene Glycol Precipitation ◽  
Dsdna Antibodies

A new method for the detection by flow cytometry of anti-double-stranded DNA antibodies and of circulating immune complexes (IC) containing endogenous DNA (IC-eDNA) is described. From each serum sample, two samples were taken, one was used to detect IC-eDNA. The other to detect anti-DNA antibodies was incubated with calf thymus DNA. ICs were isolated by polyethylene glycol precipitation or by cryoprecipitation, after which immunoglobulins were labeled with FITC-conjugated anti-human globulin. Serum samples from 63 systemic lupus erythematosus (SLE) patients, 32 incomplete lupus, and 87 control patients were tested. Detection of anti-dsDNA antibodies by flow cytometry had a diagnostic sensitivity and specificity almost comparable to routine tests, the fluorescent enzyme immunoassay EliA™-dsDNA test, and the ultrasensitive Crithidia luciliae indirect immunofluorescence test. In 21 (33%) out of 63 SLE serum samples, IC-eDNA was detected. In these samples, free anti-dsDNA antibodies were hardly detectable or undetectable by flow cytometry or by routine tests. When anti-DNA antibodies are neutralized by endogenous DNA and can no longer be detected by routine tests, the serologic diagnosis and the follow-up of relapses in patients with SLE is compromised. To overcome this obstacle, we propose an accessible solution: the detection of circulating IC-eDNA by flow cytometry.

scholarly journals Immune deposit and vasculopathy in metabolic-active lung tissues of patients with pulmonary tuberculosis

2021 ◽  
Author(s):  
Hui Ma ◽  
Lin Wang ◽  
Zilu Wen ◽  
Xinchun Chen ◽  
Haiying Liu ◽  
...  
Keyword(s):  
Gene Expression ◽  
Systemic Lupus Erythematosus ◽  
Inflammatory Response ◽  
Pulmonary Tuberculosis ◽  
Autoimmune Diseases ◽  
Metabolic Activity ◽  
Lupus Erythematosus ◽  
Immune Complexes ◽  
Systemic Lupus ◽  
Dsdna Antibodies

ABSTRACTMetabolic activity in pulmonary lesion is associated with disease severity and relapse risk in tuberculosis. However, the nature of the metabolic activity associated with tuberculosis in humans remains unclear. Previous works indicate that tuberculosis bears resemblance transcriptionally with systemic lupus erythematosus in peripheral blood, except that the plasma cell component was absent in tuberculosis. Here we reported that the missing transcriptional component was present within the metabolic active tissues in the lung of patients with sputum culture-negative tuberculosis, within which increased levels of circulating immune complexes and anti-dsDNA antibodies were found relative to nearby non-metabolic active tissues. Histological examination revealed specific vascular deposition of immune complexes, neutrophil extracellular traps, and vascular necrosis in the metabolic-active tissue. Thus, tuberculosis-initiated metabolic activity was associated with hyperactive antibody responses and vascular pathology, and shared features with systemic lupus erythematosus and other autoimmune diseases. We discussed these observations in the context of earlier literatures demonstrating that similar effects could be induced in humans and animal models by complete freund’s adjuvant, the most potent antibody response inducer ever reported. Our small case series, if verified in a larger size study, might help inform host-directed therapies to alleviate disease progression and augment treatment efficacy.IMPORTANCEIn patients with pulmonary tuberculosis, lung tissues were destroyed by a hyperactive inflammatory response towards M. tuberculosis. The mechanisms underlying the inflammatory response are still poorly understood. Using 18F-FDG avidity as a surrogate marker of inflammation, we have identified that hyper-inflamed tissues possessed features associated with systemic lupus erythematosus: gene expression signatures of plasma cell and immunoglobulins and increased levels of anti-dsDNA antibodies, immune deposits, and vasculopathy. This observation might suggest an explanation to why patients with tuberculosis share more gene expression signatures with autoimmune diseases than infectious diseases and why they are more likely to develop autoimmune diseases. Defining the inflammatory responses at the lesion could help inform host-directed therapies to intervene disease progression or even accelerate cure.

Immune complexes: characteristics, clinical correlations, and interpretive approaches in the clinical laboratory.

1982 ◽  
Vol 28 (6) ◽  
pp. 1259-1271 ◽  
Author(s):  
S E Ritzmann ◽  
J C Daniels
Keyword(s):  
Immune Complex ◽  
Lupus Erythematosus ◽  
Immune Complexes ◽  
Complex Disease ◽  
Clinical Medicine ◽  
Clinical Laboratory ◽  
Therapeutic Monitoring ◽  
Serum Samples ◽  
Complement Components ◽  
Antigen Antibody

Abstract Immune-complex-mediated injury is thought to play a role in diseases such as rheumatoid arthritis, systemic lupus erythematosus, serum sickness, various infectious diseases, and malignancies. With increased appreciation of the biological and pathological significance of circulating immune complexes has come efforts to develop appropriate techniques for identifying and measuring them. Common approaches exploit such phenomena as the attachment of complement components to antigen-antibody complexes, the presence of specialized receptors for immune complexes at the surface of cells, and the ability of rheumatoid factor to bind with immune complexes. This variety of assay systems for immune complexes has yielded abstruse results in numerous human pathological conditions. Unfortunately, these results seldom correlate with one another in a given disease. Thus, use of a panel of immune complex assays has been recommended. Indirect consequences of immune complex disease may still be appraised and evaluated with some confidence in clinical medicine: measurements of C3 and C4, cryoglobulins, serum viscosity, and turbidity of serum samples. Measurement of immune complexes may be useful in diagnosis, prognosis, and therapeutic monitoring, but it is the characterization of immune complexes that holds the greatest potential for better understanding of disease mechanisms.

scholarly journals Interaction between anti-DNA and anti-DNA-binding protein autoantibodies in cryoglobulins from sera of patients with systemic lupus erythematosus.

1986 ◽  
Vol 164 (4) ◽  
pp. 1029-1042 ◽  
Author(s):  
W H Reeves ◽  
N Chiorazzi
Keyword(s):  
Systemic Lupus Erythematosus ◽  
Dna Binding ◽  
Lupus Erythematosus ◽  
Immune Complexes ◽  
Protein Complex ◽  
Binding Protein ◽  
Dna Binding Protein ◽  
Systemic Lupus ◽  
Variable Regions ◽  
Dna Antibodies

We have previously shown that sera from some patients with SLE and related disorders contain autoantibodies to a DNA-binding protein complex designated p70/p80. The present study shows that anti-p70/p80 autoantibodies are frequently accompanied by anti-DNA antibodies and cryoglobulins. When the cryoglobulins were isolated, they were found to be specifically enriched in both anti-p70/p80 and anti-DNA activities. The anti-p70/p80 and anti-DNA antibodies were found to be distinct populations of autoantibodies rather than a single crossreactive species, since they could be separated from one another by chromatography on DNA-cellulose. Certain human anti-DNA mAbs could inhibit the binding of autoimmune polyclonal anti-p70/p80 antibodies to p70/p80, suggesting that anti-DNA antibodies might also associate with the variable regions of some anti-p70/p80 antibodies in the cryoglobulins. Binding of one murine anti-p70/p80 mAb (111-12) also was inhibited by certain human anti-DNA mAbs, but the binding of another murine mAb (162-11) to a different epitope of p70/p80 was not. These studies suggest that certain anti-DNA antibodies may interact with the variable regions of a population of anti-p70/p80 antibodies. The cryoglobulins found in the sera containing both anti-p70/p80 and anti-DNA antibodies may represent immune complexes consisting, in part, of idiotype and antiidiotype.

Serum BLC/CXCL13 Concentrations and Renal Expression of CXCL13/CXCR5 in Patients with Systemic Lupus Erythematosus and Lupus Nephritis

2009 ◽  
Vol 37 (1) ◽  
pp. 45-52 ◽  
Author(s):  
HUI-TING LEE ◽  
YU-MING SHIAO ◽  
TSAI-HUNG WU ◽  
WEI-SHENG CHEN ◽  
YUNG-HSIANG HSU ◽  
...  
Keyword(s):  
Systemic Lupus Erythematosus ◽  
Flow Cytometry ◽  
B Cells ◽  
Lupus Nephritis ◽  
Lupus Erythematosus ◽  
B Lymphocyte ◽  
Venous Blood ◽  
Systemic Autoimmune Disease ◽  
Systemic Lupus ◽  
Serum Samples

Objective.Systemic lupus erythematosus (SLE) is a prototype of systemic autoimmune disease in which cytokines such as B lymphocyte chemoattractant (BLC, or CXC motif ligand 13, CXCL13) may play important roles in pathogenesis. We investigated the implications of CXCL13 in SLE and lupus nephritis.Methods.Serum samples from 425 patients with SLE and 106 healthy control individuals were analyzed for the concentration of CXCL13 by ELISA. Tissue expression of CXCL13 and its corresponding receptor CXCR5 were observed in lupus kidney. The CXCR5-bearing B cells in SLE patients were analyzed by flow cytometry.Results.Serum levels of CXCL13 were higher in SLE patients compared to controls. SLE patients with lupus nephritis or positive anti-dsDNA antibodies had significantly higher serum CXCL13 levels. The peripheral venous blood B cells that bear CXCR5 were more abundant in SLE patients as detected by flow cytometry. CXCR5 and CXCL13 were highly expressed in the renal cortex from patients with lupus nephritis.Conclusions.Our results suggest that BLC/CXCL13 as well as its corresponding receptor, CXCR5, may play important roles in the pathogenesis of SLE and in lupus nephritis.

Performance analysis of automated evaluation of Crithidia luciliae-based indirect immunofluorescence tests in a routine setting – strengths and weaknesses

2017 ◽  
Vol 56 (1) ◽  
Author(s):  
Wymke Hormann ◽  
Melanie Hahn ◽  
Stefan Gerlach ◽  
Nicola Hochstrate ◽  
Kai Affeldt ◽  
...  
Keyword(s):  
Systemic Lupus Erythematosus ◽  
Lupus Erythematosus ◽  
Diagnostic Marker ◽  
Systemic Lupus ◽  
Diagnostic Laboratory ◽  
Automated Evaluation ◽  
Crithidia Luciliae ◽  
The Everyday ◽  
Dsdna Antibodies ◽  
Result Interpretation

AbstractBackground:Antibodies directed against dsDNA are a highly specific diagnostic marker for the presence of systemic lupus erythematosus and of particular importance in its diagnosis. To assess anti-dsDNA antibodies, theMethods:We analyzed the performance of the EUROPattern Suite with its automated fluorescence interpretation for CLIFT in a routine setting, reflecting the everyday life of a diagnostic laboratory. Three hundred and twelve consecutive samples were collected, sent to the Central Diagnostic Laboratory of the Maastricht University Medical Centre with a request for anti-dsDNA analysis over a period of 7 months.Results:Agreement between EUROPattern assay analysis and the visual read was 93.3%. Sensitivity and specificity were 94.1% and 93.2%, respectively. The EUROPattern Suite performed reliably and greatly supported result interpretation.Conclusions:Automated image acquisition is readily performed and automated image classification gives a reliable recommendation for assay evaluation to the operator. The EUROPattern Suite optimizes workflow and contributes to standardization between different operators or laboratories.

scholarly journals Features of systemic lupus erythematosus in mice injected with bacterial lipopolysaccharides: identificantion of circulating DNA and renal localization of DNA-anti-DNA complexes.

1977 ◽  
Vol 145 (5) ◽  
pp. 1115-1130 ◽  
Author(s):  
S Izui ◽  
P H Lambert ◽  
G J Fournié ◽  
H Türler ◽  
P A Miescher
Keyword(s):  
Immune Complex ◽  
Lupus Erythematosus ◽  
Immune Complexes ◽  
Basement Membranes ◽  
Circulating Dna ◽  
Dna Complexes ◽  
Renal Glomeruli ◽  
Glomerular Deposits ◽  
Dna Antibodies ◽  
Similar Density

After injection of lipopolysaccharides (LPS) in mice, there is first a release of DNA into plasma and secondly an induction of anti-DNA antibodies. The circulating DNA was purified from plasma and physico-immunochemically characterized. This DNA has a similar density to mammalian cellular DNA,is 4--6S insize, and probably represents a mixture of single-stranded DNA (SSDNA) and double-stranded DNA (DSDNA) or DSDNA with some single-stranded regions. This purified DNA was shown to react with anti-DNA antibodies which appeared as early as 3 days after a single injection of LPS in mice. In serum, DNA-anti-DNA complexes were not detected, although unidentified circulating immune complex-like material was demonstrated 5-8 days after the injection of LPS. In tissues, particularly in renal glomeruli, fine granular immune complex-type immunoglobulin deposits appeared along the glomerular capillary walls and in the mesangium 3 days after the injection of LPS. There is a direct correlation between the level of anti-DNA antibodies and the intensity of glomerular deposits and about 40% of immunoglobulins eluted from kidneys are anti-DNA antibodies, indicating that some of the immune complexes localized in kidneys are DNA-anti-DNA complexes. Based on these observations, the following hypothetical mechanism for the glomerular localization of DNA-anti-DNA complexes after the injection of LPS in mice is proposed. First, DNA, which has been released in circulating blood after injection of LPS, might bind to renal glomeruli, probably on glomerular basement membranes (GBM) through a high affinity of GBM for DNA; secondly, circulating anti-DNA antibodies, which appear later, might react with the glomerular-bound DNA and form immune complexes independently of circulating immune complexes. However, the possibility of direct deposition of immune complexes is not ruled out.

scholarly journals An electrochemical biosensor for rapid detection of anti-dsDNA antibodies in absolute scale

2018 ◽  
Author(s):  
Pablo Fagúndez ◽  
Gustavo Brañas ◽  
Justo Laíz ◽  
Juan Pablo Tosar
Keyword(s):  
Lupus Erythematosus ◽  
Electrochemical Biosensor ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Complex Samples ◽  
Absolute Scale ◽  
Laboratory Facility ◽  
Autoimmune Pathology ◽  
Assay Time ◽  
Dsdna Antibodies

AbstractAutoimmune diseases are chronic inflammatory pathologies that are characterized by the presence of antibodies against own epitopes in serum (autoantibodies). Systemic lupus erythematosus (SLE) is a common autoimmune pathology, characterized by the presence of antinuclear antibodies (ANAs). These include anti-dsDNA (α-dsDNA) antibodies, which are widely used for diagnosis and disease monitoring. Their determination is carried out by traditional techniques such as Indirect Immunofluorescence (IFI) or Enzyme Linked Immunosorbent Assay (ELISA), which are time consuming, require qualified technicians, and are not compatible with decentralized analysis outside a laboratory facility. Here, we show a sandwich-format electrochemical biosensor-based method for α-dsDNA determination in a rapid and simple manner. Total assay time is only 30 minutes and the sensor is capable of detecting 16 ng (8 μg / mL) of α-dsDNA antibodies. Using the current derived from the detection limit of the method as a cut-off, we could discriminate positive from negative serum samples with 90% sensitivity and 100% specificity. By using monoclonal antibodies for calibration curves, our results are presented in absolute scale (i.e., concentration instead of serum title) what will help to perform comparisons between methods and further improvements of this protocol. In an effort to render the sensor compatible with automation, we minimized manipulation steps without compromise of the analytical performance, even in complex samples such as serum.

Autoantibodies to insulin do appear in non-diabetic patients with autoimmune disorders: Comparison with anti-immunoglobulin antibodies and other autoimmune phenomena

1990 ◽  
Vol 122 (2) ◽  
pp. 303-308 ◽  
Author(s):  
Umberto Di Mario ◽  
Riccardo Perfetti ◽  
Emanuela Anastasi ◽  
Giovanna Contreas ◽  
Laura Crisà ◽  
...  
Keyword(s):  
Autoimmune Diseases ◽  
Lupus Erythematosus ◽  
Immune Complexes ◽  
Type I Diabetes ◽  
Primary Hypothyroidism ◽  
Insulin Autoantibodies ◽  
Diabetic Patients ◽  
Graves Disease ◽  
Type I ◽  
Serum Samples

Abstract Insulin- and anti-immunoglobulin-antibodies have been recently reported in pre-diabetic subjects: the former has been proposed as a predictive marker of Type I diabetes in non-diabetic-subjects. To evaluate the diabetes-related specificity of these antibodies, the presence of insulin autoantibodies, using a recently developed and highly sensitive competitive radioimmune assay, and of anti-immunoglobulin antibodies together with that of immune complexes and of other autoantibodies has been investigated in patients with organ- or non-organ-specific autoimmune diseases. One hundred and eleven serum samples were assayed from patients with Graves' disease, primary hypothyroidism, chronic autoimmune thyroiditis, Addison's disease, chronic autoimmune hepatitis, pernicious anemia, lupus erythematosus, and rheumatoid arthritis, together with 45 serum samples from normal subjects. From patients with autoimmune diseases, 32.4% of all sera revealed values of insulin autoantibodies above the limit of positivity (p<0.001); anti-immunoglobulin antibodies were present in 4.1% of patients (NS); immune complexes were found in 19.5% (NS) of all patients, but in 38% of patients with Graves' disease and chronic hepatitis (p<0.02). There was a trend for multiple autoantibody positivity to be associated with high levels of insulin autoantibodies (p<0.05). Thus, whereas contrary to expectation anti-immunoglobulin antibodies are not associated with non-diabetes-related autoimmune diseases, increased humoral immunoresponsiveness to endogenous insulin appears to be related to autoimmunity in general rather than restricted to Type I diabetes.

scholarly journals Automated Evaluation ofCrithidia luciliaeBased Indirect Immunofluorescence Tests: A Novel Application of the EUROPattern-Suite Technology

2015 ◽  
Vol 2015 ◽  
pp. 1-8 ◽  
Author(s):  
Stefan Gerlach ◽  
Kai Affeldt ◽  
Lena Pototzki ◽  
Christopher Krause ◽  
Jörn Voigt ◽  
...  
Keyword(s):  
Indirect Immunofluorescence ◽  
Lupus Erythematosus ◽  
Clinical Manifestations ◽  
Automated Evaluation ◽  
Crithidia Luciliae ◽  
Moderate Sensitivity ◽  
Major Criterion ◽  
Increasing Demand ◽  
Dsdna Antibodies

Systemic lupus erythematosus (SLE) is a severe rheumatic autoimmune disease with various clinical manifestations. Anti-dsDNA antibodies are an important immunological hallmark of SLE and their occurrence represents a major criterion for the diagnosis. Among the commonly applied test systems for determination of anti-dsDNA antibodies, the indirect immunofluorescence test (IIFT) using the flagellatedkinetoplastida Crithidia luciliaeis considered to be highly disease specific at moderate sensitivity. Since IIFT, however, is claimed to be affected by subjective interpretation and a lack of standardization, there has been an increasing demand for automated pattern interpretation of immunofluorescence reactions in recent years. Corresponding platforms are already available for evaluation of anti-nuclear antibody (ANA) IIFT on HEp-2 cells, the recommended “gold standard” for ANA screening in the diagnosis of various systemic rheumatic autoimmune diseases. For one of these systems, the “EUROPattern-Suite” computer-aided immunofluorescence microscopy (CAIFM), automated interpretation of microscopic fluorescence patterns was extended to theCrithidia luciliaebased anti-dsDNA IIFT.

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