Skip to main content
NCBI home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1987 Aug 1;166(2):539–549. doi: 10.1084/jem.166.2.539

Advanced glycosylation endproducts on erythrocyte cell surface induce receptor-mediated phagocytosis by macrophages. A model for turnover of aging cells

PMCID: PMC2189583  PMID: 3598465

Abstract

Glucose can react nonenzymatically with amino groups of proteins to form covalent Amadori products. With time these adducts undergo further rearrangements to form irreversible advanced glycosylation endproducts (AGE), which accumulate with protein age. A specific AGE, 2-(2-furoyl)- 4(5)-(2-furanyl)-1H-imidazole (FFI), has been identified on proteins in vivo. We have recently shown that a macrophage receptor specifically recognizes and internalizes proteins modified by AGE such as FFI, thus preferentially degrading senescent macromolecules. Reasoning that cellular turnover may be mediated by macrophage recognition of AGE- membrane proteins, we prepared human RBCs with FFI attached chemically. Human monocytes were incubated with either FFI-RBCs, IgG-opsonized RBCs, or PBS-treated RBCs. Erythrophagocytosis of FFI-RBCs was significantly higher than that of PBS-RBCs (55 vs. 4%; p less than 0.0025) and almost as high as that of IgG-RBCs (70%), and was competitively inhibited by AGE-BSA. AGE-RBCs were also prepared by incubating RBCs with various sugars. Human monocytes showed a 15% ingestion of glucose-RBCs, and a 26% ingestion of glucose-6-phosphate- RBCs, compared to 6% for PBS-RBCs. Similarly, diabetic mouse RBCs were phagocytosed by nearly three times more cells (21%) than normal mouse RBCs when exposed to syngeneic mouse macrophages. This phagocytosis was competitively inhibited (70%) by addition of excess AGE-BSA. The in vivo half-life of 51Cr-labeled mouse FFI-RBCs injected into syngeneic mice was reduced to 7 d, as compared to a half-life of 20 d for the controls. These data suggest that the macrophage receptor for the removal of glucose-modified proteins may also mediate the endocytosis of RBCs with AGE formed on their surface, and thus be responsible in part for the removal of some populations of aging cells.

Full Text

The Full Text of this article is available as a PDF (724.0 KB).

Selected References

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

  1. Andreassen T. T., Seyer-Hansen K., Bailey A. J. Thermal stability, mechanical properties and reducible cross-links of rat tail tendon in experimental diabetes. Biochim Biophys Acta. 1981 Oct 12;677(2):313–317. doi: 10.1016/0304-4165(81)90101-x. [DOI] [PubMed] [Google Scholar]
  2. Bailey A. J., Robins S. P., Tanner M. J. Reducible components in the proteins of human erythrocyte membrane. Biochim Biophys Acta. 1976 May 20;434(1):51–57. doi: 10.1016/0005-2795(76)90034-9. [DOI] [PubMed] [Google Scholar]
  3. Bevilacqua M. P., Amrani D., Mosesson M. W., Bianco C. Receptors for cold-insoluble globulin (plasma fibronectin) on human monocytes. J Exp Med. 1981 Jan 1;153(1):42–60. doi: 10.1084/jem.153.1.42. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bianco C., Eden A., Cohn Z. A. The induction of macrophage spreading: role of coagulation factors and the complement system. J Exp Med. 1976 Dec 1;144(6):1531–1544. doi: 10.1084/jem.144.6.1531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  6. Brownlee M., Pongor S., Cerami A. Covalent attachment of soluble proteins by nonenzymatically glycosylated collagen. Role in the in situ formation of immune complexes. J Exp Med. 1983 Nov 1;158(5):1739–1744. doi: 10.1084/jem.158.5.1739. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Brownlee M., Vlassara H., Cerami A. Nonenzymatic glycosylation and the pathogenesis of diabetic complications. Ann Intern Med. 1984 Oct;101(4):527–537. doi: 10.7326/0003-4819-101-4-527. [DOI] [PubMed] [Google Scholar]
  8. Brownlee M., Vlassara H., Cerami A. Trapped immunoglobulins on peripheral nerve myelin from patients with diabetes mellitus. Diabetes. 1986 Sep;35(9):999–1003. doi: 10.2337/diab.35.9.999. [DOI] [PubMed] [Google Scholar]
  9. Brownlee M., Vlassara H., Kooney A., Ulrich P., Cerami A. Aminoguanidine prevents diabetes-induced arterial wall protein cross-linking. Science. 1986 Jun 27;232(4758):1629–1632. doi: 10.1126/science.3487117. [DOI] [PubMed] [Google Scholar]
  10. Böyum A. Isolation of mononuclear cells and granulocytes from human blood. Isolation of monuclear cells by one centrifugation, and of granulocytes by combining centrifugation and sedimentation at 1 g. Scand J Clin Lab Invest Suppl. 1968;97:77–89. [PubMed] [Google Scholar]
  11. Chang J. C., Ulrich P. C., Bucala R., Cerami A. Detection of an advanced glycosylation product bound to protein in situ. J Biol Chem. 1985 Jul 5;260(13):7970–7974. [PubMed] [Google Scholar]
  12. DODGE J. T., MITCHELL C., HANAHAN D. J. The preparation and chemical characteristics of hemoglobin-free ghosts of human erythrocytes. Arch Biochem Biophys. 1963 Jan;100:119–130. doi: 10.1016/0003-9861(63)90042-0. [DOI] [PubMed] [Google Scholar]
  13. Gmelig-Meyling F., Waldmann T. A. Separation of human blood monocytes and lymphocytes on a continuous Percoll gradient. J Immunol Methods. 1980;33(1):1–9. doi: 10.1016/0022-1759(80)90077-0. [DOI] [PubMed] [Google Scholar]
  14. Higgins P. J., Bunn H. F. Kinetic analysis of the nonenzymatic glycosylation of hemoglobin. J Biol Chem. 1981 May 25;256(10):5204–5208. [PubMed] [Google Scholar]
  15. Kay M. M. Aging of cell membrane molecules leads to appearance of an aging antigen and removal of senescent cells. Gerontology. 1985;31(4):215–235. doi: 10.1159/000212706. [DOI] [PubMed] [Google Scholar]
  16. Kay M. M. Localization of senescent cell antigen on band 3. Proc Natl Acad Sci U S A. 1984 Sep;81(18):5753–5757. doi: 10.1073/pnas.81.18.5753. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Koenig R. J., Cerami A. Hemoglobin A Ic and diabetes mellitus. Annu Rev Med. 1980;31:29–34. doi: 10.1146/annurev.me.31.020180.000333. [DOI] [PubMed] [Google Scholar]
  18. Miller J. A., Gravallese E., Bunn H. F. Nonenzymatic glycosylation of erythrocyte membrane proteins. Relevance to diabetes. J Clin Invest. 1980 Apr;65(4):896–901. doi: 10.1172/JCI109743. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Monnier V. M., Kohn R. R., Cerami A. Accelerated age-related browning of human collagen in diabetes mellitus. Proc Natl Acad Sci U S A. 1984 Jan;81(2):583–587. doi: 10.1073/pnas.81.2.583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Paulsen E. P., Koury M. Hemoglobin AIc levels in insulin-dependent and -independent diabetes mellitus. Diabetes. 1976;25(2 Suppl):890–896. [PubMed] [Google Scholar]
  21. Peterson C. M., Jones R. L., Koenig R. J., Melvin E. T., Lehrman M. L. Reversible hematologic sequelae of diabetes mellitus. Ann Intern Med. 1977 Apr;86(4):425–429. doi: 10.7326/0003-4819-86-4-425. [DOI] [PubMed] [Google Scholar]
  22. Pongor S., Ulrich P. C., Bencsath F. A., Cerami A. Aging of proteins: isolation and identification of a fluorescent chromophore from the reaction of polypeptides with glucose. Proc Natl Acad Sci U S A. 1984 May;81(9):2684–2688. doi: 10.1073/pnas.81.9.2684. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. REYNOLDS T. M. CHEMISTRY OF NONENZYMIC BROWNING. I. THE REACTION BETWEEN ALDOSES AND AMINES. Adv Food Res. 1963;12:1–52. doi: 10.1016/s0065-2628(08)60005-1. [DOI] [PubMed] [Google Scholar]
  24. Schlepper-Schäfer J., Kolb-Bachofen V., Kolb H. Identification of a receptor for senescent erythrocytes on liver macrophages. Biochem Biophys Res Commun. 1983 Sep 15;115(2):551–559. doi: 10.1016/s0006-291x(83)80180-6. [DOI] [PubMed] [Google Scholar]
  25. Schlüter K., Drenckhahn D. Co-clustering of denatured hemoglobin with band 3: its role in binding of autoantibodies against band 3 to abnormal and aged erythrocytes. Proc Natl Acad Sci U S A. 1986 Aug;83(16):6137–6141. doi: 10.1073/pnas.83.16.6137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Singer J. A., Jennings L. K., Jackson C. W., Dockter M. E., Morrison M., Walker W. S. Erythrocyte homeostasis: antibody-mediated recognition of the senescent state by macrophages. Proc Natl Acad Sci U S A. 1986 Aug;83(15):5498–5501. doi: 10.1073/pnas.83.15.5498. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Vlassara H., Brownlee M., Cerami A. Accumulation of diabetic rat peripheral nerve myelin by macrophages increases with the presence of advanced glycosylation endproducts. J Exp Med. 1984 Jul 1;160(1):197–207. doi: 10.1084/jem.160.1.197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Vlassara H., Brownlee M., Cerami A. High-affinity-receptor-mediated uptake and degradation of glucose-modified proteins: a potential mechanism for the removal of senescent macromolecules. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5588–5592. doi: 10.1073/pnas.82.17.5588. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Vlassara H., Brownlee M., Cerami A. Novel macrophage receptor for glucose-modified proteins is distinct from previously described scavenger receptors. J Exp Med. 1986 Oct 1;164(4):1301–1309. doi: 10.1084/jem.164.4.1301. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Williams S. K., Devenny J. J., Bitensky M. W. Micropinocytic ingestion of glycosylated albumin by isolated microvessels: possible role in pathogenesis of diabetic microangiopathy. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2393–2397. doi: 10.1073/pnas.78.4.2393. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Wright S. D., Silverstein S. C. Tumor-promoting phorbol esters stimulate C3b and C3b' receptor-mediated phagocytosis in cultured human monocytes. J Exp Med. 1982 Oct 1;156(4):1149–1164. doi: 10.1084/jem.156.4.1149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Yue D. K., McLennan S., Delbridge L., Handelsman D. J., Reeve T., Turtle J. R. The thermal stability of collagen in diabetic rats: correlation with severity of diabetes and non-enzymatic glycosylation. Diabetologia. 1983 Apr;24(4):282–285. doi: 10.1007/BF00282714. [DOI] [PubMed] [Google Scholar]

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

RESOURCES