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Clinical and Experimental Immunology logoLink to Clinical and Experimental Immunology
. 1988 Sep;73(3):436–442.

Analysis of negatively charged dye-binding antibodies reactive with double-stranded DNA and heparan sulfate in serum from patients with rheumatic diseases.

S Aotsuka 1, M Okawa-Takatsuji 1, M Kinoshita 1, R Yokohari 1
PMCID: PMC1541765  PMID: 2974766

Abstract

Antibodies to double-stranded (ds) DNA are characteristically present in serum from patients with systemic lupus erythematosus (SLE). Recently, anti-dsDNA antibodies have been shown to have the capacity to react with a diversity of molecules with repeating negative charges. Using the anionic dye Cibacron blue F3GA, bound to crosslinked agarose, we analysed the nature of antibodies capable of reacting with this dye in serum samples from patients with various rheumatic diseases. The dye-antibody complex could easily be split by eluting with solutions of increasing ionic strength, suggesting that the interaction is ionic in nature. Pepsin-digested F(ab')2 antibodies retained the capacity to bind Cibacron blue, confirming that the binding occurred via antigen-binding sites on the antibody molecule. The eluates obtained from dye-ligand chromatography of active SLE sera contained antibodies to both dsDNA and heparan sulfate, while those of sera from patients with other non-SLE rheumatic diseases contained antibodies only against heparan sulfate. Furthermore, the dye-ligand eluates of sera from patients with active SLE and other non-SLE rheumatic diseases were found to contain increased amounts of IgG. In one patient with SLE, levels of antibodies to dsDNA and heparan sulfate, and the amounts of total IgG in dye-ligand eluates, were shown to be correlated with disease activity.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aarden L. A., Lakmaker F., Feltkamp T. E. Immunology of DNA. I. The influence of reaction conditions on the Farr assay as used for the detection of anti-ds DNA. J Immunol Methods. 1976;10(1):27–37. doi: 10.1016/0022-1759(76)90004-1. [DOI] [PubMed] [Google Scholar]
  2. Aotsuka S., Okawa M., Ikebe K., Yokohari R. Measurement of anti-double-stranded DNA antibodies in major immunoglobulin classes. J Immunol Methods. 1979;28(1-2):149–162. doi: 10.1016/0022-1759(79)90337-5. [DOI] [PubMed] [Google Scholar]
  3. Böhme H. J., Kopperschläger G., Schulz J., Hofmann E. Affinity chromatography of phosphofructokinase using Cibacron blue F3G-A. J Chromatogr. 1972 Jun 28;69(1):209–214. doi: 10.1016/s0021-9673(00)83103-9. [DOI] [PubMed] [Google Scholar]
  4. De Maeyer-Guignard J., Thang M. N., De Maeyer E. Binding of mouse interferon to polynucleotides. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3787–3790. doi: 10.1073/pnas.74.9.3787. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Emlen W., Burdick G. Purification of DNA antibodies using cibacron blue F3GA affinity chromatography. J Immunol Methods. 1983 Aug 26;62(2):205–215. doi: 10.1016/0022-1759(83)90248-x. [DOI] [PubMed] [Google Scholar]
  6. Faaber P., Capel P. J., Rijke G. P., Vierwinden G., van de Putte L. B., Koene R. A. Cross-reactivity of anti-DNA antibodies with proteoglycans. Clin Exp Immunol. 1984 Mar;55(3):502–508. [PMC free article] [PubMed] [Google Scholar]
  7. Faaber P., Rijke T. P., van de Putte L. B., Capel P. J., Berden J. H. Cross-reactivity of human and murine anti-DNA antibodies with heparan sulfate. The major glycosaminoglycan in glomerular basement membranes. J Clin Invest. 1986 Jun;77(6):1824–1830. doi: 10.1172/JCI112508. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Koike T., Tomioka H., Kumagai A. Antibodies cross-reactive with DNA and cardiolipin in patients with systemic lupus erythematosus. Clin Exp Immunol. 1982 Nov;50(2):298–302. [PMC free article] [PubMed] [Google Scholar]
  9. Kumar S. A., Beach T. A., Dickerman H. W. Effect of Cibacron blue F3GA on oligonucleotide binding site of estradiol--receptor complexes of mouse uterine cytosol. Proc Natl Acad Sci U S A. 1979 May;76(5):2199–2203. doi: 10.1073/pnas.76.5.2199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Meyer R. R., Glassberg J., Scott J. V., Kornberg A. A temperature-sensitive single-stranded DNA-binding protein from Escherichia coli. J Biol Chem. 1980 Apr 10;255(7):2897–2901. [PubMed] [Google Scholar]
  11. Okudaira H., Terada E., Ogita T., Aotsuka S., Yokohari R. Anti-double strand (ds) DNA antibody formation by NZB/W (F1) spleen cells in a microculture system detected by solid phase radioimmunoassay. J Immunol Methods. 1981;41(2):201–213. doi: 10.1016/0022-1759(81)90243-x. [DOI] [PubMed] [Google Scholar]
  12. Poikonen K., Oka M., Möttönen T., Jokinen I., Arvilommi H. Synthesis of IgM, IgG and IgA in rheumatoid arthritis. Ann Rheum Dis. 1982 Dec;41(6):607–611. doi: 10.1136/ard.41.6.607. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Pollard K. M., Webb J. Partial purification of anti-DNA antibodies from systemic lupus erythematosus serum by dye-ligand chromatography. J Immunol Methods. 1982 Oct 15;54(1):81–94. doi: 10.1016/0022-1759(82)90116-8. [DOI] [PubMed] [Google Scholar]
  14. Shoenfeld Y., Rauch J., Massicotte H., Datta S. K., André-Schwartz J., Stollar B. D., Schwartz R. S. Polyspecificity of monoclonal lupus autoantibodies produced by human-human hybridomas. N Engl J Med. 1983 Feb 24;308(8):414–420. doi: 10.1056/NEJM198302243080802. [DOI] [PubMed] [Google Scholar]
  15. Shoenfeld Y., Rauch J., Massicotte H., Datta S. K., André-Schwartz J., Stollar B. D., Schwartz R. S. Polyspecificity of monoclonal lupus autoantibodies produced by human-human hybridomas. N Engl J Med. 1983 Feb 24;308(8):414–420. doi: 10.1056/NEJM198302243080802. [DOI] [PubMed] [Google Scholar]
  16. Thompson S. T., Stellwagen E. Binding of Cibacron blue F3GA to proteins containing the dinucleotide fold. Proc Natl Acad Sci U S A. 1976 Feb;73(2):361–365. doi: 10.1073/pnas.73.2.361. [DOI] [PMC free article] [PubMed] [Google Scholar]

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