Skip to main content
NCBI home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1986 May 1;163(5):1064–1084. doi: 10.1084/jem.163.5.1064

Antibodies to basement membrane heparan sulfate proteoglycans bind to the laminae rarae of the glomerular basement membrane (GBM) and induce subepithelial GBM thickening

PMCID: PMC2188093  PMID: 2939168

Abstract

Antibodies specific for the core protein of basement membrane HSPG (Mr = 130,000) were administered to rats by intravenous injection, and the pathologic consequences on the kidney were determined at 3 min to 2 mo postinjection. Controls were given antibodies against gp330 (the pathogenic antigen of Heymann nephritis) or normal rabbit IgG. The injected anti-HSPG(GBM) IgG disappeared rapidly (by 1 d) from the circulation. The urinary excretion of albumin increased in a dose- dependent manner during the first 4 d, was increased 10-fold at 1-2 mo, but remained moderate (mean = 12 mg/24 h). By immunofluorescence the anti-HSPG(GBM) was seen to bind rapidly (by 3 min) to all glomerular capillaries, and by immunoperoxidase staining the anti-HSPG was seen to bind exclusively to the laminae rarae of the GBM where it remained during the entire 2-mo observation period. C3 was detected in glomeruli immediately after the injection (3 min), where it bound exclusively to the lamina rara interna; the amount of C3 bound increased up to 2 h but decreased rapidly thereafter, and was not detectable after 4 d. Mononuclear and PMN leukocytes accumulated in glomerular capillaries, adhered to the capillary wall, and extended pseudopodia through the endothelial fenestrae to contact in the LRI of the GBM where the immune deposits and C3 were located. At 1 wk postinjection, staining for C3 reappeared in the glomeruli of some of the rats, and by this time most of the rats, including controls injected with normal rabbit IgG, had circulating anti-rabbit IgG (by ELISA) and linear deposits of rat IgG along the GBM (by immunofluorescence). Beginning at 9 d, there was progressive subepithelial thickening of the GBM which in some places was two to three times its normal width. This thickening was due to the laying down of a new layer of basement membrane-like material on the epithelial side of the GBM, which gradually displaced the old basement membrane layers toward the endothelium. The results show that the core proteins of this population of basement membrane HSPG (Mr = 130,000), which are ubiquitous components of basement membranes, are exposed to the circulation and can bind anti-HSPG(GBM) IgG in the laminae rarae of the GBM. Binding of these antibodies to the GBM leads to changes (C3 deposition, leukocyte adherence, moderate proteinuria, GBM thickening) considered typical of the acute phase of anti-GBM glomerulonephritis. Antibody binding interferes with the normal turnover of the GBM, presumably by affecting the biosynthesis and/or degradation of basement membrane components.

Full Text

The Full Text of this article is available as a PDF (4.7 MB).

Selected References

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

  1. ANDRES G. A., MORGAN C., HSU K. C., RIFKIND R. A., SEEGAL B. C. Electron microscopic studies of experimental nephritis with ferritin-conjugated antibody. The basement membranes and cisternae of visceral epithelial cells in nephritic rat glomeruli. J Exp Med. 1962 May 1;115:929–936. doi: 10.1084/jem.115.5.929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Abrahamson D. R., Caulfield J. P. Proteinuria and structural alterations in rat glomerular basement membranes induced by intravenously injected anti-laminin immunoglobulin G. J Exp Med. 1982 Jul 1;156(1):128–145. doi: 10.1084/jem.156.1.128. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Alousi M. A., Post R. S., Heymann W. Experimental autoimmune nephrosis in rats. Morphogenesis of the glomerular lesion: immunohistochemical and electron microscopic studies. Am J Pathol. 1969 Jan;54(1):47–71. [PMC free article] [PubMed] [Google Scholar]
  4. Barry D. N., Bowness J. M. Identification and turnover of glycosaminoglycans in rat kidneys. Can J Biochem. 1975 Jun;53(6):713–720. doi: 10.1139/o75-098. [DOI] [PubMed] [Google Scholar]
  5. Brown W. J., Farquhar M. G. The mannose-6-phosphate receptor for lysosomal enzymes is concentrated in cis Golgi cisternae. Cell. 1984 Feb;36(2):295–307. doi: 10.1016/0092-8674(84)90223-x. [DOI] [PubMed] [Google Scholar]
  6. Butkowski R. J., Wieslander J., Wisdom B. J., Barr J. F., Noelken M. E., Hudson B. G. Properties of the globular domain of type IV collagen and its relationship to the Goodpasture antigen. J Biol Chem. 1985 Mar 25;260(6):3739–3747. [PubMed] [Google Scholar]
  7. Cohen M. P., Surma M. L. In vivo biosynthesis and turnover of 35S-labeled glomerular basement membrane. Biochim Biophys Acta. 1982 Jun 16;716(3):337–340. doi: 10.1016/0304-4165(82)90025-3. [DOI] [PubMed] [Google Scholar]
  8. Couser W. G., Jermanovich N. B., Belok S., Stilmant M. M., Hoyer J. R. Effect of aminonucleoside nephrosis on immune complex localization in autologous immune complex nephropathy in rats. J Clin Invest. 1978 Mar;61(3):561–572. doi: 10.1172/JCI108967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Druet P., Bariety J., Bellon B., Laliberte F. Nephrotoxic serum nephritis in the rat: ultrastructural localization of nephrotoxic rabbit antibodies using peroxidase-labeled conjugates. Lab Invest. 1972 Aug;27(2):157–164. [PubMed] [Google Scholar]
  10. Fillit H., Damle S. P., Gregory J. D., Volin C., Poon-King T., Zabriskie J. Sera from patients with poststreptococcal glomerulonephritis contain antibodies to glomerular heparan sulfate proteoglycan. J Exp Med. 1985 Feb 1;161(2):277–289. doi: 10.1084/jem.161.2.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gustafsson J. E., Uzqueda H. R. The influence of citrate and phosphate on the Mancini single radial immunodiffusion technique and suggested improvements for the determination of urinary albumin. Clin Chim Acta. 1978 Dec 15;90(3):249–257. doi: 10.1016/0009-8981(78)90264-4. [DOI] [PubMed] [Google Scholar]
  12. Hoedemaeker P. J., Feenstra K., Nijkeuter A., Arends A. Ultrastructural localization of heterologous nephrotoxic antibody in the glomerular basement membrane of the rat. Lab Invest. 1972 May;26(5):610–613. [PubMed] [Google Scholar]
  13. Kanwar Y. S. Biophysiology of glomerular filtration and proteinuria. Lab Invest. 1984 Jul;51(1):7–21. [PubMed] [Google Scholar]
  14. Kanwar Y. S., Farquhar M. G. Anionic sites in the glomerular basement membrane. In vivo and in vitro localization to the laminae rarae by cationic probes. J Cell Biol. 1979 Apr;81(1):137–153. doi: 10.1083/jcb.81.1.137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kanwar Y. S., Farquhar M. G. Presence of heparan sulfate in the glomerular basement membrane. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1303–1307. doi: 10.1073/pnas.76.3.1303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kanwar Y. S., Hascall V. C., Farquhar M. G. Partial characterization of newly synthesized proteoglycans isolated from the glomerular basement membrane. J Cell Biol. 1981 Aug;90(2):527–532. doi: 10.1083/jcb.90.2.527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kerjaschki D., Farquhar M. G. Immunocytochemical localization of the Heymann nephritis antigen (GP330) in glomerular epithelial cells of normal Lewis rats. J Exp Med. 1983 Feb 1;157(2):667–686. doi: 10.1084/jem.157.2.667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kerjaschki D., Farquhar M. G. The pathogenic antigen of Heymann nephritis is a membrane glycoprotein of the renal proximal tubule brush border. Proc Natl Acad Sci U S A. 1982 Sep;79(18):5557–5561. doi: 10.1073/pnas.79.18.5557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Koffler D., Biesecker G., Noble B., Andres G. A., Martinez-Hernandez A. Localization of the membrane attack complex (MAC) in experimental immune complex glomerulonephritis. J Exp Med. 1983 Jun 1;157(6):1885–1905. doi: 10.1084/jem.157.6.1885. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mahieu P., Lambert P. H., Miescher P. A. Detection of anti-glomerular basement membrane antibodies by radioimmunological technique. Clinical application in human nephropathies. J Clin Invest. 1974 Jul;54(1):128–137. doi: 10.1172/JCI107734. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mannik M., Agodoa L. Y., David K. A. Rearrangement of immune complexes in glomeruli leads to persistence and development of electron-dense deposits. J Exp Med. 1983 May 1;157(5):1516–1528. doi: 10.1084/jem.157.5.1516. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Markwell M. A. A new solid-state reagent to iodinate proteins. I. Conditions for the efficient labeling of antiserum. Anal Biochem. 1982 Sep 15;125(2):427–432. doi: 10.1016/0003-2697(82)90025-2. [DOI] [PubMed] [Google Scholar]
  23. Marquardt H., Wilson C. B., Dixon F. J. Isolation and immunological characterization of human glomerular basement membrane antigens. Kidney Int. 1973 Feb;3(2):57–65. doi: 10.1038/ki.1973.12. [DOI] [PubMed] [Google Scholar]
  24. Martinez-Hernandez A., Amenta P. S. The basement membrane in pathology. Lab Invest. 1983 Jun;48(6):656–677. [PubMed] [Google Scholar]
  25. McLean I. W., Nakane P. K. Periodate-lysine-paraformaldehyde fixative. A new fixation for immunoelectron microscopy. J Histochem Cytochem. 1974 Dec;22(12):1077–1083. doi: 10.1177/22.12.1077. [DOI] [PubMed] [Google Scholar]
  26. Miettinen A., Törnroth T., Tikkanen I., Virtanen I., Linder E. Heymann nephritis induced by kidney brush border glycoproteins. Lab Invest. 1980 Dec;43(6):547–555. [PubMed] [Google Scholar]
  27. Milici A. J., L'Hernault N., Palade G. E. Surface densities of diaphragmed fenestrae and transendothelial channels in different murine capillary beds. Circ Res. 1985 May;56(5):709–717. doi: 10.1161/01.res.56.5.709. [DOI] [PubMed] [Google Scholar]
  28. Mynderse L. A., Hassell J. R., Kleinman H. K., Martin G. R., Martinez-Hernandez A. Loss of heparan sulfate proteoglycan from glomerular basement membrane of nephrotic rats. Lab Invest. 1983 Mar;48(3):292–302. [PubMed] [Google Scholar]
  29. Price R. G., Spiro R. G. Studies on the metabolism of the renal glomerular basement membrane. Turnover measurements in the rat with the use of radiolabeled amino acids. J Biol Chem. 1977 Dec 10;252(23):8597–8602. [PubMed] [Google Scholar]
  30. Stow J. L., Sawada H., Farquhar M. G. Basement membrane heparan sulfate proteoglycans are concentrated in the laminae rarae and in podocytes of the rat renal glomerulus. Proc Natl Acad Sci U S A. 1985 May;82(10):3296–3300. doi: 10.1073/pnas.82.10.3296. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Tokuyasu K. T., Maher P. A., Singer S. J. Distributions of vimentin and desmin in developing chick myotubes in vivo. I. Immunofluorescence study. J Cell Biol. 1984 Jun;98(6):1961–1972. doi: 10.1083/jcb.98.6.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Unanue E. R., Dixon F. J. Experimental glomerulonephritis: immunological events and pathogenetic mechanisms. Adv Immunol. 1967;6:1–90. doi: 10.1016/s0065-2776(08)60521-0. [DOI] [PubMed] [Google Scholar]
  33. Vogt A., Rohrbach R., Shimizu F., Takamiya H., Batsford S. Interaction of cationized antigen with rat glomerular basement membrane: in situ immune complex formation. Kidney Int. 1982 Jul;22(1):27–35. doi: 10.1038/ki.1982.128. [DOI] [PubMed] [Google Scholar]
  34. Walker F. The origin, turnover and removal of glomerular basement-membrane. J Pathol. 1973 Jul;110(3):233–244. doi: 10.1002/path.1711100306. [DOI] [PubMed] [Google Scholar]
  35. Wick G., Müller P. U., Timpl R. In vivo localization and pathological effects of passively transferred antibodies to type IV collagen and laminin in mice. Clin Immunol Immunopathol. 1982 Jun;23(3):656–665. doi: 10.1016/0090-1229(82)90328-2. [DOI] [PubMed] [Google Scholar]
  36. Wieslander J., Bygren P., Heinegård D. Antiglomerular basement membrane antibody: antibody specificity in different forms of glomerulonephritis. Kidney Int. 1983 Jun;23(6):855–861. doi: 10.1038/ki.1983.106. [DOI] [PubMed] [Google Scholar]
  37. Wieslander J., Bygren P., Heinegård D. Isolation of the specific glomerular basement membrane antigen involved in Goodpasture syndrome. Proc Natl Acad Sci U S A. 1984 Mar;81(5):1544–1548. doi: 10.1073/pnas.81.5.1544. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Yaar M., Foidart J. M., Brown K. S., Rennard S. I., Martin G. R., Liotta L. The Goodpasture-like syndrome in mice induced by intravenous injections of anti-type IV collagen and anti-laminin antibody. Am J Pathol. 1982 Apr;107(1):79–91. [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES