Membrane Properties of Senescent and Carrier Human Erythrocytes

  • Maria A. Castellana
  • Maria R. De Renzis
  • Giampiero Piccinini
  • Giampaolo Minetti
  • Claudio Seppi
  • Cesare Balduini
  • Augusta Brovelli
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 326)

Abstract

Studies about human red cell senescence have shown that the viability and post-transfusion survival of red cells is related to the structure of their plasma membrane.1 In an attempt to analyze the survival potential in the circulation of red cells manipulated for loading with drugs or biosubstances, we addressed our investigation to the identification of new parameters useful to describe membrane characteristics of red cells with a decreased life expectancy. In this study we have analyzed membrane properties of young, middle-aged, and senescent red cells, and compared them with those of red cells manipulated for loading, in order to discern the membrane structural lesions leading to a decreased survival potential. Removal from the circulation of senescent red cells seems to be triggered by the binding of autologous antibodies2 recognizing band 3 (B3) protein3–5, α-galactosyl groups, probably belonging to glycolipids6, and other epitopes not yet defined.5 Although the role played by autoantibodies in the removal of senescent cells has not been completely elucidated, their presence on the surface of senescent erythrocytes focused on plasma membrane studies about cell aging and raised questions about the mechanisms leading to the expression on the cell surface of the senescence antigens. Among the processes modifying the structure of membrane components and described to occur during red cell senescence, oxidation seems to play an important role.7–11 Therefore we have analyzed the oxidative state of membrane proteins in young, middle-aged and senescent normal red cells and tried to relate it with the functional activity of B3 protein12, considering that the involvement of B3 in the expression of the senescence antigen has been recognized by different authors.3–5,11 The same investigation was carried out on red cells submitted to hypotonic dialysis and resealed. The aim of this investigation was to identify steps of cell loading processes producing cell suffering and decrease of the survival potential, in order to prevent or minimize the cellular damage with appropriate protocols.

Keywords

Glutathione Electrophoresis Pyruvate Methionine Thiol 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    M.R. Clark, Senescence of red blood cells: progress and problems, Physiol. Rev. 68:503 (1988).PubMedGoogle Scholar
  2. 2.
    M.M.B. Kay, Mechanism of removal of senescent cells by human macrophages ‘in situ’, Proc. Natl. Acad. Sci. USA 72:3521 (1975).PubMedCrossRefGoogle Scholar
  3. 3.
    M.M.B. Kay, Localization of senescent cell antigen on band 3, Proc. Natl. Acad. Sci. USA 81:5753 (1984).PubMedCrossRefGoogle Scholar
  4. 4.
    H.U. Lutz, R. Flepp and G. Stringaro-Wipf, Naturally occurring autoantibodies to exoplasmic and cryptic regions of band 3 protein, the major integral membrane protein of human red blood cells, J. Immunol. 133:2610 (1984).PubMedGoogle Scholar
  5. 5.
    M.P. Sorette, U. Galili and M.R. Clark, Comparison of serum anti-band 3 and anti-Gal antibody binding to density-separated human red blood cells, Blood 77:628 (1991).PubMedGoogle Scholar
  6. 6.
    U. Galili, B.A. Macher, J. Buehler and S.B. Shohet, Human natural anti-alpha-galactosyl IgG. II. The specific recognition of alpha(1–3)-linked galactose residues, J. Exp. Med. 162:573 (1985).PubMedCrossRefGoogle Scholar
  7. 7.
    G.J. Johnson, D.W. Allen, T.P. Flynn, B. Finkel and J.G. White, Decreased survival in vivo of diamide-incubated dog erythrocytes, J. Clin. Invest. 66:955 (1980).PubMedCrossRefGoogle Scholar
  8. 8.
    M.M.B. Kay, G.J.C.G.M. Bosman, S.S. Shapiro, A. Bendich and P.S. Bassel, Oxidation as a possible mechanism of cellular aging: vitamin E deficiency causes premature aging and IgG binding to erythrocytes, Proc. Natl. Acad. Sci. USA 83:2463 (1986).PubMedCrossRefGoogle Scholar
  9. 9.
    P. Arese, F. Bussolino, R. Flepp, P. Stammler, S. Fasler and H.U. Lutz, Diamide enhances phagocytosis of human red cell in a complement-and anti band 3 antibody-dependent process, Biomed. Biochim. Acta 46:S84 (1987).PubMedGoogle Scholar
  10. 10.
    M. Beppu, A. Mizukami, M. Nagoya and K. Kikugawa, Binding of anti-band 3 auto-antibody to oxidatively damaged erythrocytes, J. Biol. Chem. 265:3226 (1990).PubMedGoogle Scholar
  11. 11.
    P.S. Low, Interaction of native and denatured hemoglobins with band 3: consequences for erythrocyte structure and function, in: “Red Blood Cell Membranes”, P. Agre and J.C. Parker, eds, M. Dekker, New York, (1989).Google Scholar
  12. 12.
    D. Jay and L. Cantley, Structural aspects of the red cell anion exchange protein, Ann. Rev. Biochem. 55:511(1986).Google Scholar
  13. 13.
    E. Beutler, C. West and K. G. Blume, The removal of leukocytes and platelets from whole blood, J. Lab. Clin. Med. 88:328 (1976).PubMedGoogle Scholar
  14. 14.
    C. Seppi, M.A. Castellana, G. Minetti, G. Piccinini, C. Balduini and A. Brovelli, Evidence for membrane protein oxidation during in vivo aging of human erythrocytes Mech. Ageing Dev. 57:247 (1991).PubMedCrossRefGoogle Scholar
  15. 15.
    J. R. Murphy, Influence of temperature and method of centrifugation on the separation of erythrocytes, J. Lab. Clin. Med. 82:334 (1973).PubMedGoogle Scholar
  16. 16.
    W. J. Griffiths, The determination of creatine in body fluids and muscle, and of phosphocreatine in muscle, using the autoanalyzer, Clin. Chim. Acta 9:210 (1964).PubMedCrossRefGoogle Scholar
  17. 17.
    J. Fehr and M. Knob, Comparison of red cell creatine level and reticulocyte count in appraising the severity of hemolytic processes, Blood 53:966 (1979).PubMedGoogle Scholar
  18. 18.
    V. T. Marchesi and J. E. Palade, The localization of Mg-Na-K-activated adenosine triphosphatase on red cell ghost membranes, J. Cell Biol. 35:385 (1967).PubMedCrossRefGoogle Scholar
  19. 19.
    C. Ropars, M. Chassaigne, M.C. Villeral, G. Avenard, C. Hurel and C. Nicolau, Resealed red blood cells as a new blood transfusion product, in: “Red Cells as Carrier for Drugs”, J.R. De Loach and U. Sprandel, eds., Karger, Basel (1985).Google Scholar
  20. 20.
    K. Yamamoto, T. Sehne and Y. Kanaoka, Fluorescent thiol reagents - Fluorescent tracer method for protein SH groups using N-(7-dimethylamino-4-methyl coumarinyl) maleimide. An application to the proteins separated by SDS-polyacrylamide gel electrophoresis, Anal. Biochem. 79:83 (1977).PubMedCrossRefGoogle Scholar
  21. 21.
    E. Nigg, M. Kessler and R.J. Cherry, Labelling of human erythrocyte membranes with eosin probes used for protein diffusion measurements. Inhibition of anion transport and photo-oxidative inactivation of acetylcholinesterase, Biochim. Biophys. Acta 550:328 (1979).PubMedCrossRefGoogle Scholar
  22. 22.
    C.E. Cobb and A.H. Beth, Identification of the eosynil-5-maleimide reaction site on the human erythrocyte anion exchange protein: overlap with the reaction sites of other chemical probes, Biochemistry 29:8283 (1990).PubMedCrossRefGoogle Scholar
  23. 23.
    T. Chiba, Y. Sato and Y. Suzuki, Characterization of eosin 5-isothiocyanate binding site in band 3 protein of the human erythrocyte, Biochim. Biophys. Acta 897:14 (1987).PubMedCrossRefGoogle Scholar
  24. 24.
    M.K. Ho and G. Guidotti, A membrane protein from human erythrocytes involved in anion exchange, J. Biol. Chem. 250:675 (1975).PubMedGoogle Scholar
  25. 25.
    U. K. Laemmli, Cleavage of structural proteins during the assembly of the head of bacteriophage T4, Nature 227: 680 (1970).PubMedCrossRefGoogle Scholar
  26. 26.
    O. H. Lowry, N. J. Rosebrough, A. L. Fan and R. J. Randall, Protein measurement with the Folin phenol reagent, J. Biol. Chem. 193:265 (1951).PubMedGoogle Scholar
  27. 27.
    E. Beutler, The preparation of red cells for assay, in: “Red cell metabolism - A Manual of Biochemical Methods” 3rd edition, Grune and Stratton, New York (1984).Google Scholar
  28. 28.
    E. Beutler, Reduced glutathione (GSH), in: “Red cell metabolism - A Manual of Biochemical Methods” 3rd edition, Grune and Stratton, New York (1984).Google Scholar
  29. 29.
    F. Tietze, Enzymic method for quantitative determination of nanogram amounts of total and oxidized glutathione, Anal. Biochem. 27:502 (1969).PubMedCrossRefGoogle Scholar
  30. 30.
    K.J.A. Davies and A.L. Goldberg, Proteins damaged by oxygen radicals are rapidly degraded in extracts of red blood cells, J. Biol. Chem. 262:8227 (1987).PubMedGoogle Scholar
  31. 31.
    A. Brovelli, C. Seppi, A.M. Castellana, M.R. De Renzis, A. Blasina and C. Balduini, Oxidative lesion to membrane proteins in senescent erythrocytes, Biomed. Biochim. Acta 49:S218 (1990).PubMedGoogle Scholar
  32. 32.
    G. Bartosz, Erythrocyte aging: physical and chemical membrane changes, Gerontology 37:33 (1991).PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1992

Authors and Affiliations

  • Maria A. Castellana
    • 1
  • Maria R. De Renzis
    • 1
  • Giampiero Piccinini
    • 1
  • Giampaolo Minetti
    • 1
  • Claudio Seppi
    • 1
  • Cesare Balduini
    • 1
  • Augusta Brovelli
    • 1
  1. 1.Dipartimento di BiochimicaUniversità degli Studi di PaviaPaviaItaly

Personalised recommendations