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. 1998 Oct;57(10):606–613. doi: 10.1136/ard.57.10.606

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

G Servais 1, M Guillaume 1, N Dumarey 1, J Duchateau 1
PMCID: PMC1752489  PMID: 9893572

Abstract

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.

 Keywords: anti-nuclear antibodies; systemic lupus erythematosus, immunofluorescence, cytoplasmic membrane associated DNA

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Figure 1  .

Figure 1  

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Immunofluorescence patterns on methanol-fixed Wil2 cells. (A) pattern "0"= absence of any significant fluorescence; (B) pattern "1"= punctuate membrane fluorescence; (C) pattern "2"= continuous membrane peripheral fluorescence.

Figure 2  .

Figure 2  

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Comparative histogram. Distribution of fluorescence patterns in SLE compared with normal serum samples at three different dilutions: 1/2, 1/20, 1/40.

Selected References

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

  1. Albani S., Massa M., Viola S., Pellegrini G., Martini A. Antibody reactivity against single stranded DNA of various species in normal children and in children with diffuse connective tissue diseases. Autoimmunity. 1990;8(1):77–80. doi: 10.3109/08916939008998436. [DOI] [PubMed] [Google Scholar]
  2. Bennett R. M., Cornell K. A., Merritt M. J., Bakke A. C., Hsu P. H., Hefeneider S. H. Autoimmunity to a 28-30 kD cell membrane DNA binding protein: occurrence in selected sera from patients with SLE and mixed connective tissue disease (MCTD). Clin Exp Immunol. 1991 Dec;86(3):374–379. doi: 10.1111/j.1365-2249.1991.tb02940.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bennett R. M., Cornell K. A., Merritt M. J., Bakke A. C., Mourich D., Hefeneider S. H. Idiotypic mimicry of a cell surface DNA receptor: evidence for anti-DNA antibodies being a subset of anti-anti-DNA receptor antibodies. Clin Exp Immunol. 1992 Dec;90(3):428–433. doi: 10.1111/j.1365-2249.1992.tb05863.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bennett R. M., Davis J., Campbell S., Portnoff S. Lactoferrin binds to cell membrane DNA. Association of surface DNA with an enriched population of B cells and monocytes. J Clin Invest. 1983 Mar;71(3):611–618. doi: 10.1172/JCI110807. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bennett R. M., Davis J., Merritt M. Anti-DNA antibodies react with DNA expressed on the surface of monocytes and B lymphocytes. J Rheumatol. 1986 Aug;13(4):679–685. [PubMed] [Google Scholar]
  6. Bennett R. M., Gabor G. T., Merritt M. M. DNA binding to human leukocytes. Evidence for a receptor-mediated association, internalization, and degradation of DNA. J Clin Invest. 1985 Dec;76(6):2182–2190. doi: 10.1172/JCI112226. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Emlen W., Holers V. M., Arend W. P., Kotzin B. Regulation of nuclear antigen expression on the cell surface of human monocytes. J Immunol. 1992 May 15;148(10):3042–3048. [PubMed] [Google Scholar]
  8. Hefeneider S. H., Bennett R. M., Pham T. Q., Cornell K., McCoy S. L., Heinrich M. C. Identification of a cell-surface DNA receptor and its association with systemic lupus erythematosus. J Invest Dermatol. 1990 Jun;94(6 Suppl):79S–84S. doi: 10.1111/1523-1747.ep12875170. [DOI] [PubMed] [Google Scholar]
  9. Hefeneider S. H., McCoy S. L., Morton J. I., Bakke A. C., Cornell K. A., Brown L. E., Bennett R. M. DNA binding to mouse cells is mediated by cell-surface molecules: the role of these DNA-binding molecules as target antigens in murine lupus. Lupus. 1992 May;1(3):167–173. doi: 10.1177/096120339200100308. [DOI] [PubMed] [Google Scholar]
  10. Jacob L., Lety M. A., Bach J. F., Louvard D. Human systemic lupus erythematosus sera contain antibodies against cell-surface protein(s) that share(s) epitope(s) with DNA. Proc Natl Acad Sci U S A. 1986 Sep;83(18):6970–6974. doi: 10.1073/pnas.83.18.6970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Jacob L., Viard J. P., Allenet B., Anin M. F., Slama F. B., Vandekerckhove J., Primo J., Markovits J., Jacob F., Bach J. F. A monoclonal anti-double-stranded DNA autoantibody binds to a 94-kDa cell-surface protein on various cell types via nucleosomes or a DNA-histone complex. Proc Natl Acad Sci U S A. 1989 Jun;86(12):4669–4673. doi: 10.1073/pnas.86.12.4669. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kaplan J. G., Rosenberg R., Reinitz E., Buchbinder S., Schaumburg H. H. Invited review: peripheral neuropathy in Sjogren's syndrome. Muscle Nerve. 1990 Jul;13(7):570–579. doi: 10.1002/mus.880130703. [DOI] [PubMed] [Google Scholar]
  13. Kubota T., Kanai Y., Miyasaka N. Interpretation of the cross-reactivity of anti-DNA antibodies with cell surface proteins: the role of cell surface histones. Immunol Lett. 1990 Jan;23(3):187–193. doi: 10.1016/0165-2478(90)90190-2. [DOI] [PubMed] [Google Scholar]
  14. Kuo M. T., Meinke W., Saunders G. F. Localization of cytoplasmic-membrane-associated DNA in human chromosomes. Proc Natl Acad Sci U S A. 1975 Dec;72(12):5004–5006. doi: 10.1073/pnas.72.12.5004. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lerner R. A., Meinke W., Goldstein D. A. Membrane-associated DNA in the cytoplasm of diploid human lymphocytes. Proc Natl Acad Sci U S A. 1971 Jun;68(6):1212–1216. doi: 10.1073/pnas.68.6.1212. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Meinke W., Hall M. R., Goldstein D. A., Kohne D. E., Lerner R. A. Physical properties of cytoplasmic membrane-associated DNA. J Mol Biol. 1973 Jun 25;78(1):43–56. doi: 10.1016/0022-2836(73)90427-0. [DOI] [PubMed] [Google Scholar]
  17. Moyer M. P. The association of DNA and RNA with membranes. Int Rev Cytol. 1979;61:1–61. doi: 10.1016/s0074-7696(08)61994-4. [DOI] [PubMed] [Google Scholar]
  18. Mécheri S., Dannecker G., Dennig D., Poncet P., Hoffmann M. K. Anti-histone autoantibodies react specifically with the B cell surface. Mol Immunol. 1993 Apr;30(6):549–557. doi: 10.1016/0161-5890(93)90029-b. [DOI] [PubMed] [Google Scholar]
  19. Okudaira K., Yoshizawa H., Williams R. C., Jr Monoclonal murine anti-DNA antibody interacts with living mononuclear cells. Arthritis Rheum. 1987 Jun;30(6):669–678. doi: 10.1002/art.1780300610. [DOI] [PubMed] [Google Scholar]
  20. Pisetsky D. S. Anti-DNA antibodies in systemic lupus erythematosus. Rheum Dis Clin North Am. 1992 May;18(2):437–454. [PubMed] [Google Scholar]
  21. Ramirez F., Searles R. B., Williams R. C., Jr Interactions of IgG from SLE patients with peripheral blood mononuclear cells and adherent cell populations. Rheumatol Int. 1988;8(1):15–20. doi: 10.1007/BF00541345. [DOI] [PubMed] [Google Scholar]
  22. Raz E., Ben-Bassat H., Davidi T., Shlomai Z., Eilat D. Cross-reactions of anti-DNA autoantibodies with cell surface proteins. Eur J Immunol. 1993 Feb;23(2):383–390. doi: 10.1002/eji.1830230213. [DOI] [PubMed] [Google Scholar]
  23. Reid B. L., Charlson A. J. Cytoplasmic and cell surface deoxyribonucleic acids with consideration of their origin. Int Rev Cytol. 1979;60:27–52. doi: 10.1016/s0074-7696(08)61258-9. [DOI] [PubMed] [Google Scholar]
  24. Rekvig O. P. Intrinsic cell membrane antigens recognized by antichromatin autoantibodies. The membrane antigens do not derive from the nucleus. Scand J Immunol. 1989 Jan;29(1):7–13. doi: 10.1111/j.1365-3083.1989.tb01093.x. [DOI] [PubMed] [Google Scholar]
  25. Rogers J. C., Kerstiens J. W. Capping of DNA on phytohemagglutinin-stimulated human lymphoblasts. J Immunol. 1981 Feb;126(2):703–705. [PubMed] [Google Scholar]
  26. Ruffatti A., Peserico A., Rondinone R., Calligaro A., Del Ross T., Ghirardello A., Germino M., Todesco S. Prevalence and characteristics of anti-single-stranded DNA antibodies in localized scleroderma. Comparison with systemic lupus erythematosus. Arch Dermatol. 1991 Aug;127(8):1180–1183. [PubMed] [Google Scholar]
  27. Smeenk R., Hylkema M. Detection of antibodies to DNA: a technical assessment. Mol Biol Rep. 1992 Nov;17(1):71–79. doi: 10.1007/BF01006401. [DOI] [PubMed] [Google Scholar]
  28. Swaak A. J., Huysen V., Nossent J. C., Smeenk R. J. Antinuclear antibody profiles in relation to specific disease manifestations of systemic lupus erythematosus. Clin Rheumatol. 1990 Mar;9(1 Suppl 1):82–94. doi: 10.1007/BF02205555. [DOI] [PubMed] [Google Scholar]
  29. Tan E. M., Cohen A. S., Fries J. F., Masi A. T., McShane D. J., Rothfield N. F., Schaller J. G., Talal N., Winchester R. J. The 1982 revised criteria for the classification of systemic lupus erythematosus. Arthritis Rheum. 1982 Nov;25(11):1271–1277. doi: 10.1002/art.1780251101. [DOI] [PubMed] [Google Scholar]

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