Characterization of Antibody that Binds In Vivo to Normal Human Red Blood Cells

  • Margaret R. Clark
  • Martin P. Sorette
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 307)


It has been well-documented that a subpopulation of normal circulating human red cells have antibody on their surfaces, and this antibody has been proposed to serve as a major mechanism for removal of senescent red cells from the circulation at the end of their lifespan (1). Antibodies of two different specificities have been reported to be enriched in high density human red cells. One is an antibody that appears to recognize an altered form of the Band 3 membrane protein (2). The other recognizes an a -galactosyl carbohydrate structure, found in small quantities in membrane glycolipids of Old World primates (3). As a basis for further understanding of the physiologic relevance of these antibodies, we have performed quantitative measurements of their contribution to in situ bound antibody on high density populations of normal human red cells. In addition, we have determined the number of binding sites of each type of antibody on different density populations that had been stripped of in situ antibody. To provide insight about the possible activity of the IgG dense cells, the IgG subclass distribution of the in situ antibody was also determined.


World Primate Density Gradient Separation Dense Cell Population Fluorescence Histogram Membrane Glycolipid 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    M. M. B. Kay, Mechanism of removal of senescent cells by human macrophages “in situ”, Proc. Natl. Acad. Sci. (USA) 72:3521 (1975).CrossRefGoogle Scholar
  2. 2.
    M. M. B. Kay, Localization of senescent antigen on band 3, Proc. Natl. Acad. Sci. (USA) 81:5753 (1984).CrossRefGoogle Scholar
  3. 3.
    U. Galilli, The natural anti-Gal antibody, the B-like antigen, and human red cell aging, Blood Cells 14:205 (1988).Google Scholar
  4. 4.
    M. R. Clark, Calcium extrusion by high density human red blood cells, Blood Cells 14:119 (1988).PubMedGoogle Scholar
  5. 5.
    O. P. Rekving and K. Hannestad, Acid elution of blood group antibodies from intact erythrocytes, Vox Sang. 33:286 (1977).CrossRefGoogle Scholar
  6. 6.
    M. R. Clark, N. Mohands and S. B. Shohet, Osmotic gradient ektacytometry: comprehensive characterization of red cell volume and surface maintenance, Blood 61:899 (1983).PubMedGoogle Scholar
  7. 7.
    P. Yam, L. D. Petz and P. Spath, Detection of IgG sensitization of red cells with 125I Staphylococcal Protein A, Am. J. Hematol. 12:337 (1982).PubMedCrossRefGoogle Scholar
  8. 8.
    C. Fenner, R. R. Trout, D. T. Mason, N. Wickman and J. Coffelt, Quantitation of Coomassie Blue stained proteins in Polyacrylamide gels based on analysis of the eluted dye, Anal. Biochem. 63:595 (1975).PubMedCrossRefGoogle Scholar
  9. 9.
    A. P. Gee and J. J. Langone, Immunoassay using 125I or enzyme labeled protein A and antigen coated tubes, Anal. Biochem. 116:524 (1981).PubMedCrossRefGoogle Scholar
  10. 10.
    U. Galili, R. E. Mandrell, R. M. Hamedeh, S. B. Shohet and J. M. Griffiss, Interaction between human natural anti--galactosyl immunoglobulin G and bacteria of human flora, Infect Immun. 56:1730 (1988).PubMedGoogle Scholar
  11. 11.
    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
  12. 12.
    M. F. Likacovic, M. B. Feinstein, R. I. Shaafi and S. Perrie, Purification of stabilized band 3 protein of the human erythrocyte membrane and its reconstitution into liposomes, Biochem. USA 20:3145 (1981).CrossRefGoogle Scholar
  13. 13.
    T. J. Mueller, C. W. Jackson, M. E. Dockter and M. Morrison, Membrane skeletal alterations during in vivo mouse red cell aging. Increase in the band ratio, J. Clin. Invest. 79:492 (1987).PubMedCrossRefGoogle Scholar
  14. 14.
    E. Beutler, Isolation of the aged, Blood Cells 14:1 (1988).PubMedGoogle Scholar
  15. 15.
    M. R. Clark, L. Corash and R. H. Jensen, Density distribution of aging, transfused human red cells, Blood 74 (Suppl. 1):217a (1989).Google Scholar

Copyright information

© Plenum Press, New York 1991

Authors and Affiliations

  • Margaret R. Clark
    • 1
  • Martin P. Sorette
    • 1
  1. 1.Department of Laboratory Medicine and Cancer Research InstituteUniversity of CaliforniaSan FranciscoUSA

Personalised recommendations