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Evidence of autoantibodies to cell membrane associated DNA (cultured lymphocytes): a new specific marker for rapid identification of systemic lupus erythematosus

OBJECTIVE Autoantibodies to cell membrane associated DNA are described in systemic lupus erythematosus (SLE). The specificity of these antibodies differ from antibodies to nuclear DNA. METHODS Using indirect immunofluorescence, a specific IgG was detected giving a characteristic pattern of continuou...

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Published in:Annals of the rheumatic diseases 1998-10, Vol.57 (10), p.606-613
Main Authors: Servais, Geneviève, Guillaume, Marie-Paule, Dumarey, Nicolas, Duchateau, Jean
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Guillaume, Marie-Paule
Dumarey, Nicolas
Duchateau, Jean
description OBJECTIVE Autoantibodies to cell membrane associated DNA are described in systemic lupus erythematosus (SLE). The specificity of these antibodies differ from antibodies to nuclear DNA. METHODS Using indirect immunofluorescence, a specific IgG was detected giving a characteristic pattern of continuous peripheral membrane fluorescence on cultured B-lymphocytes. RESULTS This pattern was observed in 53 of 80 serum samples of SLE patients but absent in the serum samples of the control populations: 15 rheumatoid arthritis, 38 ankylosing spondylarthritis, 17 non-inflammatory osteopenic patients, and 224 blood donors. In 34 Sjögren syndrome’s patients one only showed a positive test. The cmDNA specificity of these antibodies was confirmed by pattern extinction with DNAse but not RNase or protease pre-treatment of the cells. IgG to cmDNA, separated by absorption/elution from purified cmDNA immobilised on DEAE-nitrocellulose reproduced the immunofluorescence pattern pictures. Extensive serum depletion of anti-double strand or single strand DNA antibodies by absorption to cellulose bound ds- or ss-DNA affected marginally the pericellular fluorescence revealing some minor cross reactivity with nuclear DNA. Moreover, in SLE patients without detectable antibody to ds-DNA, pericellular fluorescence could be visible. CONCLUSION This novel rapid immunofluorescence method may serve as an identification test of SLE patients. Given its positive (97.1%) and negative (92.9%) predictive value, sensitivity (66%) and specificity (99.5%), it improves on other diagnostic tests such as the detection of antibodies to Sm.
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The specificity of these antibodies differ from antibodies to nuclear DNA. METHODS Using indirect immunofluorescence, a specific IgG was detected giving a characteristic pattern of continuous peripheral membrane fluorescence on cultured B-lymphocytes. RESULTS This pattern was observed in 53 of 80 serum samples of SLE patients but absent in the serum samples of the control populations: 15 rheumatoid arthritis, 38 ankylosing spondylarthritis, 17 non-inflammatory osteopenic patients, and 224 blood donors. In 34 Sjögren syndrome’s patients one only showed a positive test. The cmDNA specificity of these antibodies was confirmed by pattern extinction with DNAse but not RNase or protease pre-treatment of the cells. IgG to cmDNA, separated by absorption/elution from purified cmDNA immobilised on DEAE-nitrocellulose reproduced the immunofluorescence pattern pictures. Extensive serum depletion of anti-double strand or single strand DNA antibodies by absorption to cellulose bound ds- or ss-DNA affected marginally the pericellular fluorescence revealing some minor cross reactivity with nuclear DNA. Moreover, in SLE patients without detectable antibody to ds-DNA, pericellular fluorescence could be visible. CONCLUSION This novel rapid immunofluorescence method may serve as an identification test of SLE patients. Given its positive (97.1%) and negative (92.9%) predictive value, sensitivity (66%) and specificity (99.5%), it improves on other diagnostic tests such as the detection of antibodies to Sm.</description><identifier>ISSN: 0003-4967</identifier><identifier>EISSN: 1468-2060</identifier><identifier>DOI: 10.1136/ard.57.10.606</identifier><identifier>PMID: 9893572</identifier><identifier>CODEN: ARDIAO</identifier><language>eng</language><publisher>London: BMJ Publishing Group Ltd and European League Against Rheumatism</publisher><subject>Adult ; Aged ; Aged, 80 and over ; anti-nuclear antibodies ; Antibodies, Antinuclear - blood ; Antibody Specificity ; Arthritis - diagnosis ; Autoimmune diseases ; Biological and medical sciences ; Biomarkers - blood ; Cell Membrane - immunology ; cytoplasmic membrane associated DNA ; Deoxyribonucleic acid ; Diagnosis, Differential ; Disease ; DNA ; DNA - immunology ; DNA, Single-Stranded - immunology ; Experiments ; Extended Reports ; Female ; Fluorescent Antibody Technique, Indirect ; Humans ; immunofluorescence ; Immunoglobulin G - blood ; Immunoglobulins ; Lupus ; Lupus Erythematosus, Systemic - diagnosis ; Lupus Erythematosus, Systemic - immunology ; Lymphocytes ; Lymphoma ; Male ; Medical sciences ; Methanol ; Middle Aged ; Patients ; Proteins ; Sarcoidosis. Granulomatous diseases of unproved etiology. Connective tissue diseases. Elastic tissue diseases. 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The specificity of these antibodies differ from antibodies to nuclear DNA. METHODS Using indirect immunofluorescence, a specific IgG was detected giving a characteristic pattern of continuous peripheral membrane fluorescence on cultured B-lymphocytes. RESULTS This pattern was observed in 53 of 80 serum samples of SLE patients but absent in the serum samples of the control populations: 15 rheumatoid arthritis, 38 ankylosing spondylarthritis, 17 non-inflammatory osteopenic patients, and 224 blood donors. In 34 Sjögren syndrome’s patients one only showed a positive test. The cmDNA specificity of these antibodies was confirmed by pattern extinction with DNAse but not RNase or protease pre-treatment of the cells. IgG to cmDNA, separated by absorption/elution from purified cmDNA immobilised on DEAE-nitrocellulose reproduced the immunofluorescence pattern pictures. Extensive serum depletion of anti-double strand or single strand DNA antibodies by absorption to cellulose bound ds- or ss-DNA affected marginally the pericellular fluorescence revealing some minor cross reactivity with nuclear DNA. Moreover, in SLE patients without detectable antibody to ds-DNA, pericellular fluorescence could be visible. CONCLUSION This novel rapid immunofluorescence method may serve as an identification test of SLE patients. Given its positive (97.1%) and negative (92.9%) predictive value, sensitivity (66%) and specificity (99.5%), it improves on other diagnostic tests such as the detection of antibodies to Sm.</description><subject>Adult</subject><subject>Aged</subject><subject>Aged, 80 and over</subject><subject>anti-nuclear antibodies</subject><subject>Antibodies, Antinuclear - blood</subject><subject>Antibody Specificity</subject><subject>Arthritis - diagnosis</subject><subject>Autoimmune diseases</subject><subject>Biological and medical sciences</subject><subject>Biomarkers - blood</subject><subject>Cell Membrane - immunology</subject><subject>cytoplasmic membrane associated DNA</subject><subject>Deoxyribonucleic acid</subject><subject>Diagnosis, Differential</subject><subject>Disease</subject><subject>DNA</subject><subject>DNA - immunology</subject><subject>DNA, Single-Stranded - immunology</subject><subject>Experiments</subject><subject>Extended Reports</subject><subject>Female</subject><subject>Fluorescent Antibody Technique, Indirect</subject><subject>Humans</subject><subject>immunofluorescence</subject><subject>Immunoglobulin G - blood</subject><subject>Immunoglobulins</subject><subject>Lupus</subject><subject>Lupus Erythematosus, Systemic - diagnosis</subject><subject>Lupus Erythematosus, Systemic - immunology</subject><subject>Lymphocytes</subject><subject>Lymphoma</subject><subject>Male</subject><subject>Medical sciences</subject><subject>Methanol</subject><subject>Middle Aged</subject><subject>Patients</subject><subject>Proteins</subject><subject>Sarcoidosis. 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The specificity of these antibodies differ from antibodies to nuclear DNA. METHODS Using indirect immunofluorescence, a specific IgG was detected giving a characteristic pattern of continuous peripheral membrane fluorescence on cultured B-lymphocytes. RESULTS This pattern was observed in 53 of 80 serum samples of SLE patients but absent in the serum samples of the control populations: 15 rheumatoid arthritis, 38 ankylosing spondylarthritis, 17 non-inflammatory osteopenic patients, and 224 blood donors. In 34 Sjögren syndrome’s patients one only showed a positive test. The cmDNA specificity of these antibodies was confirmed by pattern extinction with DNAse but not RNase or protease pre-treatment of the cells. IgG to cmDNA, separated by absorption/elution from purified cmDNA immobilised on DEAE-nitrocellulose reproduced the immunofluorescence pattern pictures. Extensive serum depletion of anti-double strand or single strand DNA antibodies by absorption to cellulose bound ds- or ss-DNA affected marginally the pericellular fluorescence revealing some minor cross reactivity with nuclear DNA. Moreover, in SLE patients without detectable antibody to ds-DNA, pericellular fluorescence could be visible. CONCLUSION This novel rapid immunofluorescence method may serve as an identification test of SLE patients. Given its positive (97.1%) and negative (92.9%) predictive value, sensitivity (66%) and specificity (99.5%), it improves on other diagnostic tests such as the detection of antibodies to Sm.</abstract><cop>London</cop><pub>BMJ Publishing Group Ltd and European League Against Rheumatism</pub><pmid>9893572</pmid><doi>10.1136/ard.57.10.606</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record>
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identifier ISSN: 0003-4967
ispartof Annals of the rheumatic diseases, 1998-10, Vol.57 (10), p.606-613
issn 0003-4967
1468-2060
language eng
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source Open Access: PubMed Central
subjects Adult
Aged
Aged, 80 and over
anti-nuclear antibodies
Antibodies, Antinuclear - blood
Antibody Specificity
Arthritis - diagnosis
Autoimmune diseases
Biological and medical sciences
Biomarkers - blood
Cell Membrane - immunology
cytoplasmic membrane associated DNA
Deoxyribonucleic acid
Diagnosis, Differential
Disease
DNA
DNA - immunology
DNA, Single-Stranded - immunology
Experiments
Extended Reports
Female
Fluorescent Antibody Technique, Indirect
Humans
immunofluorescence
Immunoglobulin G - blood
Immunoglobulins
Lupus
Lupus Erythematosus, Systemic - diagnosis
Lupus Erythematosus, Systemic - immunology
Lymphocytes
Lymphoma
Male
Medical sciences
Methanol
Middle Aged
Patients
Proteins
Sarcoidosis. Granulomatous diseases of unproved etiology. Connective tissue diseases. Elastic tissue diseases. Vasculitis
systemic lupus erythematosus
Tissue Fixation
Tumor Cells, Cultured
title Evidence of autoantibodies to cell membrane associated DNA (cultured lymphocytes): a new specific marker for rapid identification of systemic lupus erythematosus
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