Binding of monoclonal anti-native DNA autoantibodies to DNA of varying size and conformation

R, Ali; DerSimonian, Harout; Stollar, B. David

doi:10.1016/0161-5890(85)90065-3

Cited by 76 publications

(30 citation statements)

References 27 publications

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“…Finally, it is very unlikely that very small DNA molecules which might have run otTthegel and been lost can account for the CIE-measurable DNA. sinre molecules smaller than 34 bp would almost certainly not have been immunoprecipitabic [28], It is therefore concluded that the CIE method is in fact measuring molecules that are ON-like in structure, as visualized on PAGE, although il dt)es not necessatily follow Ihal all such visualized subfractions contribute proportionately to the lotal.…”

Section: Discussionmentioning

confidence: 99%

Haemodialysis as a model for studying endogenous plasma DNA: oligonucleosome-like structure and clearance

Rumore

Muralidhar

Lin

et al. 1992

Clinical and Experimental Immunology

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SUMMARYThe rale of clearance of exiracellular plasma DNA in man ha.s importani inipiitalions for palhogenelie mechanisms in systemic lupus erythemalosus (SLE), as well as for eeriain oihcr clinical states. Present knowledge of this parameter is derived exelusively from studies of injected, naked DNA in animals. Recent informalion indicates that the physiologic form of plasma DNA in SLE is that of oligonucleosome-like molecules rather than of naked DNA and eonsists of multimcric complexes of DNA bound to histone, probably arising from an apoplotic process. In order to study the rale at which ihese oligonucleosome-like complexes are removed from plasma and to do so in man rather than experimental animals, we exploited the observation that during haemodialysis large amounts of DNA are released, apparently within the dialysis coil, inlo the paiieni's plasma. Since this release appears lo cease promptly with termination of the procedure, it offered the potential for estimating the rate of removal of such DNA from human plasma. Moreover, if that DNA. as postulated, were shown to possess an oligonucleosome-like structure resembling that fotind endogenously in human SLE, the relevance of such information to the human disease state would be further enhanced. The present results support the conclusion that DNA released into plasma during haemodialysis possesses such an oligonucleosome-like structure. The plasma half-life of ihat DNA in man was found nol lo exceed 4 min. The highly dynamic state thus implied for extracellular endogenous plasma DNA in man has important implications for pathogenetic mechanisms dependent on dsDNA in SLE. Moreover, individuals undergoing chronic hacmodialysi,s, who are thereby exposed to a very large cumulative amount of such DNA., might serve as models for studying its long-term sequelae.

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Section: Discussionmentioning

confidence: 99%

Haemodialysis as a model for studying endogenous plasma DNA: oligonucleosome-like structure and clearance

Rumore

Muralidhar

Lin

et al. 1992

Clinical and Experimental Immunology

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“…Elucidating the mechanism of antibody-RNA interaction is crucial to understanding the probable contribution of antibodies to pathological processes in some autoimmune diseases as well as for the general understanding of structural features that determine the specificity of RNA-protein recognition+ Anti-nucleic acid antibodies play an important but still unclear role in the autoimmune disorder systemic lupus erythematosus (SLE; Gilbert et al+, 1997)+ Understanding their interaction with DNA may provide better insight into the pathogenicity of the disease (Shlomchik et al+, 1990;Chen et al+, 1995;Zouali, 1997)+ In addition to sequence and structure analysis of anti-DNA antibodies, their binding mechanisms are of special interest (Ali et al+, 1985;Stollar, 1986;Radic & Weigert, 1994)+ The major population of anti-nucleic acid antibodies is represented by DNA-specific species, but there are also antibodies directed against the left-handed helical Z-conformation (Z-DNA and Z-RNA) and against singlestranded (ss) or double-stranded (ds) RNA (Stollar, 1992)+ In contrast to DNA, which does not induce an immune response in healthy organisms, dsRNA is a relatively good antigen+ dsRNA-specific antibodies like J2 recognize structural features common to most dsRNA species+ Because many ssRNA-and dsRNA-binding proteins display apparently sequence-nonspecific interactions, anti-dsRNA antibodies may serve as a general model system to elucidate binding characteristics and binding mechanisms+ This may improve their use in diagnosis and the general investigation of viral and metabolic dsRNAs (Schönborn et al+, 1991;Lukacs, 1994)+ RNA-protein interactions have become increasingly important as a result of the perception that gene regulation at the posttranscriptional level is significant+ With the mechanisms of antisense RNA-mediated gene si-lencing and the recent discovery of RNAi as a specific inhibitor of gene expression, the role of dsRNA in cellular metabolism has become a central theme in research+ The detection of dsRNA in cells or cellular extracts is difficult because artifacts may be generated during preparation and there are only indirect methods to demonstrate the presence of genuine dsRNA molecules+ It has recently been shown that dsRNA-specific antibodies can be used to visualize dsRNA by in situ techniques (Lukacs, 1997)+ mAb J2, one of the antibodies used for in situ detection of dsRNA, has a binding preference for mixed-sequence dsRNA and poly(A)•poly(U) compared to poly(I)•poly(C) (Schönborn et al+, 1991)+ A class of proteins showing a binding behavior similar to mAb J2 is characterized by one or more dsRNAbinding domains (dsRBD) (St Johnston et al+, 1992;Finerty & Bass, 1997)+ These are domains of ;65-70 amino acids in an a-b-b-b-a configuration+ Highresolution structural information is available for four of these proteins: RNAse III from Escherichia coli (Kharrat et al+, 1995), the staufen protein from Drosophila (Bycroft et al+, 1995), human dsRNA-dependent protein kinase (PKR; …”

Section: Introductionmentioning

confidence: 99%

Determination of preferential binding sites for anti-dsRNA antibodies on double-stranded RNA by scanning force microscopy

Bonin

Oberstrass

Lukàcs

et al. 2000

RNA

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The monoclonal anti-dsRNA antibody J2 binds double-stranded RNAs (dsRNA) in an apparently sequence-nonspecific way. The mAb only recognizes antigens with double-stranded regions of at least 40 bp and its affinity to poly(A) poly(U) and to dsRNAs with mixed base pair composition is about tenfold higher than to poly(I) poly(C). Because no specific binding site could be determined, the number, the exact dimensions, and other distinct features of the binding sites on a given antigen are difficult to evaluate by biochemical methods. We therefore employed scanning force microscopy (SFM) as a method to analyze antibody-dsRNA interaction and protein-RNA binding in general. Several in vitro-synthesized dsRNA substrates, generated from the Dictyostelium PSV-A gene, were used. In addition to the expected sequence-nonspecific binding, imaging of the complexes indicated preferential binding of antibodies to the ends of dsRNA molecules as well as to certain internal sites. Analysis of 2,000 bound antibodies suggested that the consensus sequence of a preferential internal binding site is A 2 N 9 A 3 N 9 A 2 , thus presenting A residues on one face of the helix. The site was verified by site-directed mutagenesis, which abolished preferential binding to this region. The data demonstrate that SFM can be efficiently used to identify and characterize binding sites for proteins with no or incomplete sequence specificity. This is especially the case for many proteins involved in RNA metabolism.

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“…BALB/c-derived hybridoma 35D is an L-chain-only line bearing an L chain identical with V8R (14). The origins of mAb are described previously: anti-laminin IgG H50 and anti-DNA IgM 238 (19); LamH-C Tg mAb 54 and A10C (12); anti-DNA IgG H241 (20). Alkaline phosphatase-conjugated anti-isotype reagents and avidin were obtained from Pierce.…”

Section: Cell Lines and Absmentioning

confidence: 99%

κ Editing Rescues Autoreactive B Cells Destined for Deletion in Mice Transgenic for a Dual Specific Anti-Laminin Ig

Brady

Congdon

Clark

et al. 2004

The Journal of Immunology

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We explored mechanisms involved in B cell self-tolerance in a double- and triple-transgenic mouse model bearing the LamH-Cμ Ig H chain conventional transgene and a gene-targeted replacement for a functional Vκ8Jκ5 L chain gene. Whereas the H chain is known to generate anti-laminin Ig in combination with multiple L chains, the H + L Ig binds ssDNA in addition to laminin. Immune phenotyping indicates that H + L transgenic B cells are regulated by clonal deletion, receptor editing via secondary rearrangements at the nontargeted κ allele, and anergy. Collectively, the data suggest that multiple receptor-tolerogen interactions regulate autoreactive cells in the H + L double-transgenic mice. Generation of H + LL triple-transgenic mice homozygous for the targeted L chain to exclude secondary κ rearrangements resulted in profound B cell depletion with absence of mature B cells in the bone marrow. We propose that the primary tolerogen of dual reactive B cells in this model is not ssDNA, but a strongly cross-linking tolerogen, presumably basement membrane laminin, that triggers recombination-activating gene activity, L chain editing, and deletion.

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Binding of monoclonal anti-native DNA autoantibodies to DNA of varying size and conformation

Cited by 76 publications

References 27 publications

Haemodialysis as a model for studying endogenous plasma DNA: oligonucleosome-like structure and clearance

Haemodialysis as a model for studying endogenous plasma DNA: oligonucleosome-like structure and clearance

Determination of preferential binding sites for anti-dsRNA antibodies on double-stranded RNA by scanning force microscopy

κ Editing Rescues Autoreactive B Cells Destined for Deletion in Mice Transgenic for a Dual Specific Anti-Laminin Ig

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