Clinical Significance of Anti–double‐stranded Dna Antibodies Detected by a Solid Phase Enzyme Immunoassay

Miller, Thomas E.; Lahita, Robert G.; Zarro, Vincent J.; Macwilliam, Joanne; Koffler, David

doi:10.1002/art.1780240406

Cited by 35 publications

(19 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The percentage of positives in active SLE is somewhat higher than those obtained from other methods but agrees with those found by Miller et al (27) and Halbert et al (28). We found 66% of SLE sera from patients with inactive disease were positive, which was similar to the 56% found by Miller et al (27) but considerably higher than that found by other methods.…”

Section: Discussronsupporting

confidence: 89%

See 1 more Smart Citation

Enzyme immunoassay for antibodies to native DNA

Eaton

Schnneider

Schur

1983

Arthritis & Rheumatism

109

View full text Add to dashboard Cite

An enzyme immunoassay was developed to detect antibodies to native DNA; DNA coating conditions that maximized sensitivity, specificity, and reproducibility were selected. Sera of patients with systemic lupus erythematosus (SLE) were positive more frequently by this immunoassay than by the Crithidiu luciliae assay or by counterimmunoelectrophoresis. By enzyme immunoassay, 94% of sera with active SLE and 70% of sera from patients with inactive SLE were positive, as were 16% from those suspected of having SLE, and 2.5% of normal persons. Specificity for native DNA was shown for both SLE and normal sera by inhibition studies and by S1 nuclease treatment of polystyrene-bound native DNA. The enzyme immunoassay correlated more with serum hemolytic complement levels than did the other 2 assays, suggesting that it detects biologically more relevant anti-DNA antibodies than do the other 2 tests.Measurement of serum anti-DNA antibody levels has been helpful in both the diagnosis and clinical assessment of systemic lupus erythematosus (SLE) detect antibodies to native DNA (nDNA), including immunodiffusion (7), complement fixation (8,9), agglutination (lO,ll), immunofluorescence (6,12), and radioimmunoassays (13-18). The sensitivity, specificity, cost, and ease of performance of these assays vary considerably (19,20).(The enzyme immunoassay (EIA) appears to offer advantages over other assays in areas of sensitivity, reproducibility, and efficiency, without sacrificing specificity. Since the initial description by Engvall and Perlmann (21), many antigens have been bound to solid phases to detect antibodies by EIA. Most of these antigens have been proteins or polysaccharides, but the binding of nucleic acids to polystyrene has not been systematically studied. Recently, several papers have appeared describing the application of EIAs to detect antibodies to DNA (22-29). In each of these reports different conditions were used, and the success of binding nDNA to the solid phase has varied (23,25).The purpose of the present study was to develop an EIA that could detect antibodies to nDNA, including a reproducible and reliable method for binding of DNA to the solid phase; this EIA was to be both sensitive and specific. In addition, the antibodies detected by the EIA were compared with those detected by other tests to define the (biologic) quality and relevance of the antibodies analyzed by different techniques. MATERIALS AND METHODSSource of sera. Normal human sera were obtained either from hospital personnel (sera kindly provided by Mr.

show abstract

Section: Discussronsupporting

confidence: 89%

“…Several investigators have attached DNA to untreated plastics (22)(23)(24)(25)(26)(27), although some reports have claimed that nDNA is not adsorbed to untreated polystyrene or polyvinyl (34,35). Pretreatment of plastics with poly-L lysine for radioimmunoassays (3 1.32) or protamine sulfate for EIA (25) has been successfully tried.…”

Section: Discussronmentioning

confidence: 99%

Enzyme immunoassay for antibodies to native DNA

Eaton

Schnneider

Schur

1983

Arthritis & Rheumatism

109

View full text Add to dashboard Cite

show abstract

“…Calf thymus dsDNA (Worthington) was prepared by chromatography on methylated albumin Kieselguhr (1 1). The purified dsDNA antigen used for fluid-phase inhibition was considered to be operationally double-stranded because of the resistance to endonuclease treatment, the absence of reaction with rabbit anti-A or rabbit anti-T antisera (12), and the failure to inhibit human sera containing only antibodies to denatured DNA (dDNA). Denatured DNA was prepared by heating at 100°C for 10 minutes, followed by submersion in ice.…”

Section: Methodsmentioning

confidence: 99%

Reaction of antipolynucleotide antibody from systemic lupus erythematosus patient serum with double‐stranded DNA complexed to protein

Faiferman

Koffler

1987

Arthritis & Rheumatism

Self Cite

View full text Add to dashboard Cite

Antipolynucleotide antibody from systemic lupus erythematosus patient serum was found'to interact with common determinants of double-stranded DNA (dsDNA) complexed to protein within nucleosomes, and with protein-free denatured DNA. In contrast, purified dsDNA isolated from nucleosomes manifested decreased reactivity with antipolynucleotide antibody. These data suggest that immunoassays using denatured DNA antigen substrate are useful for identifying a broad spectrum of antipolynucleotide antibodies that are reactive with dsDNA complexed to protein.The presence of high-avidity antibody that is reactive with double-stranded DNA (dsDNA) is one of the major diagnostic criteria for systemic lupus erythematosus (SLE) (1). Increased titers of this antibody have also been closely correlated with flares of disease activity (2). Protein-free dsDNA is used as the antigen substrate in most immunoassays that detect or quantitate this antibody. However, in vivo, dsDNA exists as a complex with proteins, such as histone. Inasmuch as local protein-DNA interaction has been shown to alter helical conformation (3-9, one would expect differences in the reaction of antipolynucleotide antibody from SLE serum with purified dsDNA

show abstract

“…Whether DNA is an antigenic stimulus in SLE has been a subject of controversy (4,5), not only because some anti-DNA antibodies cross-react with antigens irrelevant to DNA, but also because other antigens such as cardiolipin and Klebsiella may give rise to antibodies cross-reacting with DNA (6)(7)(8)(9)(10)(11)(12)(13). Likewise, the presence of anti-DNA antibodies may or may not correlate with disease activity (14)(15)(16)(17). Despite these paradoxes, antinucleic acid antibodies in general, and anti-native DNA (n-DNA)' in particular, are still the hallmark of SLE (18,19).…”

Section: Introductionmentioning

confidence: 99%