Reaction of systemic lupus erythematosus antinative DNA antibodies with native DNA fragments from 20 to 1,200 base pairs.

Papalian, Michael M.; Lafer, Eileen M.; Wong, Richard; Stollar, B D

doi:10.1172/jci109690

Cited by 98 publications

(36 citation statements)

References 33 publications

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“…To study the biological properties of immune complexes containing DNA, factors such as DNA size, antigen-antibody molar ratios, and DNA configuration (ss vs. ds) should be defined (15). Only by using well defined complexes can the effect of each of these parameters on the in vivo behavior and pathogenicity of DNA-anti-DNA immune complexes be ascertained.…”

Section: Discussionmentioning

confidence: 99%

Clearance of circulating DNA-anti-DNA immune complexes in mice.

Emlen¹,

Mannik²

1982

The Journal of Experimental Medicine

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Immune complexes composed of DNA and antibodies to DNA contribute to tissue injury in systemic lupus erythematosus (SLE) (1, 2). High titers of antibodies to both single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA) are associated with the disease (2, 3), and antibodies to DNA have been eluted from the kidneys of SLE patients (4, 5). Direct demonstration of circulating DNA-anti-DNA immune complexes has been reported (6, 7), but not confirmed (8).Whereas the fate of circulating immun e complexes such as HSA-anti-HSA has been well characterized (9), the behavior of immune complexes containing DNA has not been studied. DNA is cleared from the circulation extremely rapidly, faster than large-latticed complexes (10). Furthermore, in vitro DNA binds to glomerular and skin basement membranes (11). These unique features of DNA might also influence the fate of immune complexes containing DNA as the antigen. In this study, immune complexes were prepared in vitro with ssDNA and antibodies to DNA from a patient with SLE. The clearance and organ localization of these immune complexes was studied in normal mice and compared with the clearance of heat aggregated immunoglobulin (IgG), serving as a model for immune complexes. Our results demonstrate that the clearance of ssDNA-anti-DNA immune complexes parallels the clearance of DNA alone and is much more rapid than the clearance of aggregated IgG. Materials and MethodsAntibodies to DNA. Serum from a patient with active SLE, kindly provided by Dr. J. V. Jones of the University of Chicago, was precipitated with 50% saturated ammonium sulphate and resuspended in borate buffer (0.2 M borate, 0.15 M NaCI; pH 8.0). This material was radiolabeled with 125I by the iodine monochloride method (12), and free 125I was removed by dialysis (final sp act 30,000 epm//~g). This preparation, containing IgM and IgG, was gel filtered over Sepharose 4B (Pharmacia Fine Chemicals, Div. of Pharmaeia Inc., Piscataway, N J) and the included IgG peak was pooled (see Figure 1 A). Reehromatography of this peak showed no excluded protein.ssDNA. Calf thymus DNA (Worthington Biochemical Corp., Freehold, N J) was heat denatured and labeled with laxI according to the method of Commerford (13). After dialysis to remove free xa'I, the DNA was passed over Sepharose 4B, and the excluded material was pooled (Fig. 1 B). Analysis of this DNA on 2% agarose gel electrophoresis revealed a mean mol wt of 640,000 with a range of 400,000-800,000.Preparation of Immune Complexes. I0 mg of 125I IgG was mixed with 20/~g of lalI ssDNA and incubated at 37°C for 30 rain. This mixture (2 ml) was gel filtered on a 50-ml Sepharose 4B column, and the 125I and 131I counts per 0.5 ml fraction were determined (Fig. 1C).

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Section: Discussionmentioning

confidence: 99%

Clearance of circulating DNA-anti-DNA immune complexes in mice.

Emlen¹,

Mannik²

1982

The Journal of Experimental Medicine

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“…The antigens were calf thymus nDNA and dDNA, RNA, poly I, poly(dT) (at a concentration 2.5 pdml), and purified cardiolipin micelles (20 pghl). The DNA was prepared as previously described (19). The double-stranded nature of the nDNA was assured by treatment with S1 nuclease.…”

Section: Methodsmentioning

confidence: 99%

Multiple serologic reactions and their relationship to clinical activity in systemic lupus erythematosus

Isenberg

Shoenfeld

Schwartz

1984

Arthritis & Rheumatism

106

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“…Calf thymus DNA, obtained from Worthington Biochemicals Corp., Freehold, N. J., was purified further as described (20). For denaturation, DNA was heated at 100°C for 10 min and quickly chilled in an ice-water bath.…”

Section: Methodsmentioning

confidence: 99%

“…Helical DNA fragments of 90-110 base pairs were prepared as described previously (20). Oligonucleotides averaging 20 residues in length were prepared from a pancreatic DNase digest of calf thymus DNA.…”

Section: Methodsmentioning

confidence: 99%

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Secondary Structure in Denatured DNA is Responsible for Its Reaction with Antinative DNA Antibodies of Systemic Lupus Erythematosus Sera

Stollar¹,

Papalian²

1980

J. Clin. Invest.

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A B S T R A C T Experiments were designed to determine the basis for the strong competitive reaction of denatured DNA with systemic lupus erythematosus (SLE) antinative DNA antibodies. Secondary structure in denatured DNA was reflected in hyperchromicity upon heating and in multiphase kinetics ofits digestion by Si nuclease. Partial digestion by Si nuclease completely eliminated the ability of denatured DNA to react with antidenatured DNA antibodies, but not its ability to react with SLE sera. Sl nuclease-resistant cores were isolated from extensively digested denatured DNA. These cores had secondary structure, including some stable fold-back helical regions. The cores, from 20 to several hundred base pairs in size, competed with native DNA for binding by SLE sera. Other experiments measured reactions of denatured DNA under conditions that affected its secondary structure content. Its competitive activity decreased as temperature was increased from 00 to 37°C, whereas the activity of native DNA was not altered in this temperature range. With DNA pieces of 90-110 base pairs, native fragments were much more effective than the denatured fragments, in which stable helical structure is less likely to occur than in high molecular weight denatured DNA. Competitive assays with mononucleotides, oligonucleotides, homopolymers, and RNA-DNA hybrids also indicated that two strands of polydeoxyribonucleotide were required for optimal reactions with these SLE serum antibodies. The antibodies can measure stable helical regions in denatured DNA; they may also stabilize short helical regions that occur in an equilibrium of conformational forms.

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Reaction of systemic lupus erythematosus antinative DNA antibodies with native DNA fragments from 20 to 1,200 base pairs.

Abstract: A B S T R A C T Double-stranded DNA fragments of varying sizes were isolated and tested for binding to systemic lupus erythematosus (SLE) antinative DNA antibodies. Fragments of 20-25, 40-50, 90-110, and 160-180

Cited by 98 publications

References 33 publications

Clearance of circulating DNA-anti-DNA immune complexes in mice.

Clearance of circulating DNA-anti-DNA immune complexes in mice.

Multiple serologic reactions and their relationship to clinical activity in systemic lupus erythematosus

Secondary Structure in Denatured DNA is Responsible for Its Reaction with Antinative DNA Antibodies of Systemic Lupus Erythematosus Sera

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