Abstract
Erythrocytes from young and old human donors were separated according to age-density on Stractan gradients. Old donors had more low age-density (young) erythrocytes than did young donors. Levels of IgG bound to old and young erythrocytes were determined by ELISA. Erythrocytes from old donors bore higher levels of IgG on their erythrocytes (123±55 IgG molecules per young RBC and 196±43 IgG per old RBC) than did those from young donors (58±15 IgG per young RBC and 98±20 IgG per old RBC). In an in vitro erythrophagocytosis assay, young and old erythrocytes from old donors and old erythrocytes from young donors were shown to be recognized and phagocytosed by lymphokine activated human peripheral blood monocytes. Young erythrocytes from young donors were not phagocytosed in this assay. The in vitro erythrophagocytosis of erythrocytes from old and young donors can be specifically blocked by ß-galactoside but not α-galactoside sugars. This phagocytosis is not blocked by Protein-G which specifically blocks Fc-gamma mediated erythrophagocytosis of Rh-D+ erythrocytes coated with IgG anti-Rh-D antibodies, ß-galactoside and α-galactoside sugars have no inhibitory effect on erythrophagocytosis mediated by IgG anti-Rh-D antibodies coating Rh-D+ erythrocytes. It thus appears that erythrophagocytosis of young and old erythrocytes from old donors and old erythrocytes from young donors are all mediated by a lectin-like receptor on the monocytes which recognizes ß-galactoside-like sugar moiety on the erythrocytes rather than by recognition of IgG on the erythrocyte and an Fc receptor on the macrophage. It also appears that the membrane of both young and old erythrocytes of old donors are marked for phagocytosis whereas only the old erythrocytes from young donors are so marked.
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References
L. M. Corash, S. Piomelli, H. Chia Chan, C. Seaman and E. Gross, Separation of erythrocytes according to age on a simplified density gradient, J. Clin. Med. 84:147 (1974).
G. D. Bennet and M. M. B. Kay, Homeostatic removal of senescent murine erythrocytes by splenic macrophages, Exp. Hematol. 9:297 (1981).
D. Danon and Y. Marikovsky, Determination of density distribution of red cell population, J. Lab. & Clin. Med. 64:668 (1964).
J. R. Murphy, Influence of temperature and method of centrifugation on the separation of erythrocytes, J. Lab. Clin. Med. 82:334 (1973).
M. R. Clark, Senescence of red blood cells: progress and problems, Physiol. Rev. 68:503 (1988).
G. A. Glass, H. Gershon, D. Gershon, The effect of donor and cell age on several characteristics of rat erythrocytes, Exp. Hematol. 11:987 (1983).
E. C. Abraham, J. F. Taylor and C. A. Lang, Influence of mouse age and erythrocyte age on glutathione metabolism, Bioch. J. 174:819 (1978).
M. Magnani, L. Rossi, V. Stocchi, L. Cucchiarini, G. Piacentini and G. Fornaini, Effect of age on some properties of mice erythrocytes, Mech. Age Dev. 42:37 (1988).
T. Vomel and D. Piatt, Lifespan of rabbit erythrocytes and activity of the reticulohistiocyte system, Mech. Age Dev. 17:267 (1981).
G. A. Glass, D. Gershon and H. Gershon, Some characteristics of the human erythrocyte as a function of donor and cell age, Exp. Hematol. 13:1122 (1985).
D. Danon, Recognition by macrophages of alterations in the membranes of old red cells and expelled nuclei, in: “Permeability and Function of Biological Membranes”, Bolis, Katchalski, Kevens, Loevenstein, Pethica, pp. 57, North-Holland (1970).
D. Aminoff, Senescence and sequestration of RBC from Circulation, in: “Cellular and Molecular Aspects of Aging; The Red Cell as a Model”, Eaton, Konzen, White, pp. 279, Liss, New York (1985).
U. Galili, I. Flechner and E. A. Rachmilewitz, A naturally occuring anti-α-galactosyl IgG recognizing senenscent human red cells, in: “Cellular and Molecular Aspects of Aging; The Red Cell as a Model”, Eaton, Konzen, White, pp. 263, Liss, New York (1985).
M. M. B. Kay, S. R. Goodman, K. Sorensen, C. F. Whitfieid, P. Wong, L. Zaki and V. Rudloff, Senescent cell antigen is immunologically related to band3, Proc. Natl. Acad. Sci. USA 80:1631 (1983).
H. U. Lutz and G. Stringaro-Wipe, Senescent red cell-bound IgG is attached to band 3 protein. Biomed. Biochim. Acta 42:S117 (1983).
K. Schluter and D. Drenckhan, Co-clustering of denatured hemoglobin with band 3: its role in binding of autoantibodies against band 3 to abnormal and aged erythrocytes, Proc. Natl. Sci. USA 83:6137 (1986).
J. J. Fidler, Macrophages and metastasis — a biological approach to cancer theraphy, Cancer Res. 45:4714 (1985).
D. Danon, L. Goldstein, Y. Marikovsky and E. Skutelsky, Use of cationized ferritin as a label of negative charge on cell surfaces, J Ultrastruct Res 38:500 (1972).
D. Aminoff, M. A. Ghalambor and C. J. Henric, Gost, galactose oxidase and sialyl transferase, substrate and receptor sites in erythrocyte senescence, in: “Erythrocyte Membrane 2: Recent Clinical and Experimental Advances”, Kruckergerg, Eaton, Brewer, pp. 269 Liss, New York (1981).
M. Kay, Role of physiologic autoantibody in the removal of senescent human red cells, J. Supramolecular Structure 9:555 (1978).
H. U. Lutz, F. Bussolino, R. Flepp, S. Fasler, P. Stammler, M. D. Kazatchkine and P. Arese, Naturally occurring anti-band-3 antibodies and complement together mediate phagocytosis of oxidatively stressed human erythrocytes, Proc. Natl. Acad. Sci. USA 84:7368 (1987).
I. O. Szymanski, P. R. Odgren, N. L. Fortier and L. M. Snyder, Red blood cell associated IgG in normal and pathologic states, Blood 55:48 (1980).
M. O. Jeje, M. A. Blajchman, K. Steeves, P. Horsewood and J. G. Kelton, Quantitation of red cell-associated IgG using an immunoradiometric assay, Transfusion 24:473 (1984).
E. Sheiban and H. Gershon, The development of an ELISA for the determination of in situ bound IgG on erythrocytes of normal donors and specific blocking of an IgG dependent erythrophagocytosis assay by Protein-G. Submitted for publication.
E. Sheiban and H. Gershon, Mechanism of recognition and sequestration of young and old erythrocytes from young and old donors. Submitted for publication.
G. C. Bosma, R. P. Custer and M. J. Bosma, A severe combined immunodeficiency in the mouse, Nature 301:527 (1983).
M. R. Lieber, J. E. Hesse, S. Lewis, G. C. Bosma, N. Rosenberg, K. Mizuichi, M. J. Bosma and M. G. Gilbert, The defect in murine SCID: Joining of signal sequences but not coding segments in V(D)J recombination, Cell 55:7 (1988).
M. G. Kim, W. Shuler, M. J. Bosma and K. B. Marcu, Abnormal recombinant of Igh D and J gene segmants in transformed pre-B cells of SCID mice. J. Immunol. 141:1341 (1988).
K. Okajaki, S. Nishikawa and H. Sakano, Aberrant immunoglobulin gene rearrangement in SCID mouse bone marrow cells. J. Immunol. 141:1348 (1988).
B. A. Malynn, T. K. Blackwell, G. M. Fulop, G. A. Rathburn, A. J. W. Furley, P. Ferrier, L. B. Heinke, R. A. Phillips, G. Yancoupolos and F. W. Alt, The SCID defect affects the final step of the immunoglobulin VDJ recombinase mechanism, Cell 54:453 (1988).
K. Dorshkind, G. M. Killer, R. A. Phillips, R. G. Miller, G. C. Bosma, M. O’Toole and M. J. Bosma, Functional status of cells from lymphoid and myeloid tissues in mice with severe combined immunodeficiency disease, J. Immunol. 132:1804 (1984).
H. Gershon, Normal erythrocyte sequestration in an agammaglobulinemic host: Studies in the CB-17 SCID mouse. Submitted for publication.
E. Beutler, C. West and K. G. Blume, The removal of leukocytes and platelets from whole blood, J. Lab. Clin. Med. 88:328 (1976).
R. P. Custer, G. C. Bosma and M. J. Bosma, Severe combined immunodeficiency (SCID) in the mouse: Pathology, reconstitution, neoplasms, Am. J. Pathol. 120:468 (1985).
G. A. Glass and D. Gershon, Decreased enzymic protection and increased sensitivity to oxidative damage in erythrocytes as a function of cell and donor age, Bioch. J. 218:531 (1984).
L. Bjork and A. Kronvall, Purification and some properties of streptococcal Protein G, a novel IgG-binding reagent, J. Immunol. 133:969 (1984).
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© 1991 Plenum Press, New York
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Gershon, H., Sheiban, E. (1991). In Vitro Sequestration of Erythrocytes from Hosts of Various Ages. In: Magnani, M., De Flora, A. (eds) Red Blood Cell Aging. Advances in Experimental Medicine and Biology, vol 307. Springer, Boston, MA. https://doi.org/10.1007/978-1-4684-5985-2_30
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DOI: https://doi.org/10.1007/978-1-4684-5985-2_30
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