Skip to main content
NCBI home page
Annals of the Rheumatic Diseases logoLink to Annals of the Rheumatic Diseases
. 1996 Jan;55(1):57–62. doi: 10.1136/ard.55.1.57

Increased release of von Willebrand factor antigen from endothelial cells by anti-DNA autoantibodies.

K N Lai 1, J C Leung 1, K B Lai 1, F M Lai 1, K C Wong 1
PMCID: PMC1010083  PMID: 8572736

Abstract

OBJECTIVE--To determine whether antibodies to double stranded DNA (anti-dsDNA) have a pathogenic role in systemic lupus erythematosus (SLE). METHODS--IgG was purified from 17 patients with SLE (median anti-dsDNA titre 1212 IU/ml) and nine healthy controls (median titre 40 IU/ml). Anti-dsDNA depleted polyclonal IgG (median anti-dsDNA titre 17 IU/ml) was also prepared from sera of the 17 patients by affinity chromatography on a DNA cellulose column. Binding to antiendothelial cell antibodies (AECA) and expression of von Willebrand factor (VWF) antigen by cultured human umbilical vein endothelial cells (HUVECs) were studied by flow cytometry. RESULTS--The percentage of HUVECs binding to AECA or expressing VWF was greater for cells incubated with IgG from patients with SLE than for cells incubated with control IgG, though values did not reach statistical significance; nevertheless, HUVECs incubated with IgG from patients expressed a greater mean fluorescence intensity with AECA (p = 0.0001) and greater VWF expression (p = 0.019). Both the fluorescence intensity and percentage of HUVECs binding to AECA or expressing VWF were significantly greater in HUVEC incubated with IgG containing anti-dsDNA than in those incubated with anti-dsDNA depleted IgG. The concentration of VWF in the supernatant was significantly increased in HUVECs incubated with IgG containing anti-dsDNA compared with control IgG or anti-dsDNA depleted IgG. Pretreatment of HUVECs with native DNA before incubation with IgG from lupus patients did not increase binding to AECA, or expression or release of VWF. CONCLUSIONS--Our study provides in vitro evidence that antibodies to DNA have a pathogenic role in the induction of inflammatory injury of the vascular endothelium in SLE.

Full text

PDF
57

Selected References

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

  1. Blann A. D. von Willebrand factor as a marker of injury to the endothelium in inflammatory vascular disease. J Rheumatol. 1993 Sep;20(9):1469–1471. [PubMed] [Google Scholar]
  2. Chan T. M., Frampton G., Cameron J. S. Identification of DNA-binding proteins on human umbilical vein endothelial cell plasma membrane. Clin Exp Immunol. 1993 Jan;91(1):110–114. doi: 10.1111/j.1365-2249.1993.tb03363.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cines D. B., Lyss A. P., Reeber M., Bina M., DeHoratius R. J. Presence of complement-fixing anti-endothelial cell antibodies in systemic lupus erythematosus. J Clin Invest. 1984 Mar;73(3):611–625. doi: 10.1172/JCI111251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. D'Cruz D. P., Houssiau F. A., Ramirez G., Baguley E., McCutcheon J., Vianna J., Haga H. J., Swana G. T., Khamashta M. A., Taylor J. C. Antibodies to endothelial cells in systemic lupus erythematosus: a potential marker for nephritis and vasculitis. Clin Exp Immunol. 1991 Aug;85(2):254–261. doi: 10.1111/j.1365-2249.1991.tb05714.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fournié G. J. Circulating DNA and lupus nephritis. Kidney Int. 1988 Feb;33(2):487–497. doi: 10.1038/ki.1988.25. [DOI] [PubMed] [Google Scholar]
  6. Giddings J. C., Shall L. Enhanced release of von Willebrand factor by human endothelial cells in culture in the presence of phorbol myristate acetate and interleukin 1. Thromb Res. 1987 Aug 1;47(3):259–267. doi: 10.1016/0049-3848(87)90139-3. [DOI] [PubMed] [Google Scholar]
  7. Hahn B. H., Ebling F. M. Suppression of NZB/NZW murine nephritis by administration of a syngeneic monoclonal antibody to DNA. Possible role of anti-idiotypic antibodies. J Clin Invest. 1983 Jun;71(6):1728–1736. doi: 10.1172/JCI110927. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hashimoto Y., Ziff M., Hurd E. R. Increased endothelial cell adherence, aggregation, and superoxide generation by neutrophils incubated in systemic lupus erythematosus and Felty's syndrome sera. Arthritis Rheum. 1982 Dec;25(12):1409–1418. doi: 10.1002/art.1780251204. [DOI] [PubMed] [Google Scholar]
  9. Isenberg D. A., Ehrenstein M. R., Longhurst C., Kalsi J. K. The origin, sequence, structure, and consequences of developing anti-DNA antibodies. A human perspective. Arthritis Rheum. 1994 Feb;37(2):169–180. doi: 10.1002/art.1780370204. [DOI] [PubMed] [Google Scholar]
  10. Izui S., Lambert P. H., Miescher P. A. Failure to detect circulating DNA--anti-DNA complexes by four radioimmunological methods in patients with systemic lupus erythematosus. Clin Exp Immunol. 1977 Dec;30(3):384–392. [PMC free article] [PubMed] [Google Scholar]
  11. Izui S., Lambert P. H., Miescher P. A. In vitro demonstration of a particular affinity of glomerular basement membrane and collagen for DNA. A possible basis for a local formation of DNA-anti-DNA complexes in systemic lupus erythematosus. J Exp Med. 1976 Aug 1;144(2):428–443. doi: 10.1084/jem.144.2.428. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Koffler D., Schur P. H., Kunkel H. G. Immunological studies concerning the nephritis of systemic lupus erythematosus. J Exp Med. 1967 Oct 1;126(4):607–624. doi: 10.1084/jem.126.4.607. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lai K. N., Leung J. C., Rifkin I., Lockwood C. M. Effect of anti-neutrophil cytoplasm autoantibodies on the intracellular calcium concentration of human neutrophils. Lab Invest. 1994 Feb;70(2):152–162. [PubMed] [Google Scholar]
  14. Lindsey N. J., Dawson R. A., Henderson F. I., Greaves M., Hughes P. Stimulation of von Willebrand factor antigen release by immunoglobulin from thrombosis prone patients with systemic lupus erythematosus and the anti-phospholipid syndrome. Br J Rheumatol. 1993 Feb;32(2):123–126. doi: 10.1093/rheumatology/32.2.123. [DOI] [PubMed] [Google Scholar]
  15. Paleolog E. M., Crossman D. C., McVey J. H., Pearson J. D. Differential regulation by cytokines of constitutive and stimulated secretion of von Willebrand factor from endothelial cells. Blood. 1990 Feb 1;75(3):688–695. [PubMed] [Google Scholar]
  16. Pearson J. D. Markers of endothelial perturbation and damage. Br J Rheumatol. 1993 Aug;32(8):651–652. doi: 10.1093/rheumatology/32.8.651. [DOI] [PubMed] [Google Scholar]
  17. Savage C. O., Pottinger B. E., Gaskin G., Lockwood C. M., Pusey C. D., Pearson J. D. Vascular damage in Wegener's granulomatosis and microscopic polyarteritis: presence of anti-endothelial cell antibodies and their relation to anti-neutrophil cytoplasm antibodies. Clin Exp Immunol. 1991 Jul;85(1):14–19. doi: 10.1111/j.1365-2249.1991.tb05675.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Shibata S., Sasaki T., Harpel P., Fillit H. Autoantibodies to vascular heparan sulfate proteoglycan in systemic lupus erythematosus react with endothelial cells and inhibit the formation of thrombin-antithrombin III complexes. Clin Immunol Immunopathol. 1994 Feb;70(2):114–123. doi: 10.1006/clin.1994.1018. [DOI] [PubMed] [Google Scholar]
  19. Suenaga R., Evans M., Hatfield M., Abdou N. I. Study of anti-DNA antibodies prepared by DNA cellulose or Cibacron blue chromatography. J Immunol Methods. 1986 Oct 23;93(1):131–140. doi: 10.1016/0022-1759(86)90443-6. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Tokunaga O., Fan J. L., Watanabe T. Atherosclerosis- and age-related multinucleated variant endothelial cells in primary culture from human aorta. Am J Pathol. 1989 Dec;135(6):967–976. [PMC free article] [PubMed] [Google Scholar]
  22. Tokunaga O., Watanabe T. Atherosclerosis and endothelium. Part 1. A simple method of endothelial cell culture from human atherosclerotic aorta. Acta Pathol Jpn. 1987 Apr;37(4):527–536. [PubMed] [Google Scholar]
  23. Tsai C. Y., Wu T. H., Sun K. H., Liao T. S., Lin W. M., Yu C. L. Polyclonal IgG anti-dsDNA antibodies exert cytotoxic effect on cultured rat mesangial cells by binding to cell membrane and augmenting apoptosis. Scand J Rheumatol. 1993;22(4):162–171. doi: 10.3109/03009749309099265. [DOI] [PubMed] [Google Scholar]
  24. Westphal J. R., Boerbooms A. M., Schalwijk C. J., Kwast H., De Weijert M., Jacobs C., Vierwinden G., Ruiter D. J., Van de Putte L. B., De Waal R. M. Anti-endothelial cell antibodies in sera of patients with autoimmune diseases: comparison between ELISA and FACS analysis. Clin Exp Immunol. 1994 Jun;96(3):444–449. doi: 10.1111/j.1365-2249.1994.tb06049.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Winfield J. B., Faiferman I., Koffler D. Avidity of anti-DNA antibodies in serum and IgG glomerular eluates from patients with systemic lupus erythematosus. Association of high avidity antinative DNA antibody with glomerulonephritis. J Clin Invest. 1977 Jan;59(1):90–96. doi: 10.1172/JCI108626. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Yap H. K., Ng S. C., Lee B. W., Seah C. C., Tan M. A., Choy M. Y., Jordan S. C. Modulation of MHC expression on human endothelial cells by sera from patients with systemic lupus erythematosus. Clin Immunol Immunopathol. 1993 Sep;68(3):321–326. doi: 10.1006/clin.1993.1133. [DOI] [PubMed] [Google Scholar]

Articles from Annals of the Rheumatic Diseases are provided here courtesy of BMJ Publishing Group

RESOURCES