Flow cytometric analysis of erythrocyte populations in tn syndrome blood using monoclonal antibodies to glycophorin A and the Tn antigen

Authors

  • Dr. William L. Bigbee,

    Corresponding author
    1. Sciences Division, Lawrence Livermore National Laboratory, University of California, Livermore, California 94550
    • Biomedical Sciences Division, Lawrence Livermore National Laboratory, University of California, P.O. Box 5507, L-452, Livermore, CA 94550
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  • Richard G. Langlois,

    1. Sciences Division, Lawrence Livermore National Laboratory, University of California, Livermore, California 94550
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  • Larry H. Stanker,

    1. Sciences Division, Lawrence Livermore National Laboratory, University of California, Livermore, California 94550
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  • Martin Vanderlaan,

    1. Sciences Division, Lawrence Livermore National Laboratory, University of California, Livermore, California 94550
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  • Ronald H. Jensen

    1. Sciences Division, Lawrence Livermore National Laboratory, University of California, Livermore, California 94550
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  • Supported in part by grant No. R811819-01 from the U.S. Environmental Protection Agency and by grant No. 2S07-RR05917-04 from the U.S. National Institutes of Health under the auspices of the U.S. Department of Energy by the Lawrence Livermore National Laboratory under contract No. W-7405-ENG-48.

  • DISCLAIMER. This document was prepared as an account of work sponsored by an agency of the United States Government. Neither the United States Government nor the University of California nor any of their employees, makes any warranty, express or implied, or assumes any legal liability or responsibility for the accuracy, completeness, or usefulness of any information, apparatus, product, or process disclosed, or represents that its use would not infringe privately owner rights. Reference herein to any specific commercial products, process, or service by trade name, trademark, manufacturer, or otherwise, does not necessarily constitute or imply its endorsement, recommendation, or favoring by the United States Government or the University of California. The views and opinions of authors expressed herein do not necessarily state or reflect those of the United States Government or the University of California, and shall not be used for advertising or product endorsement purposes.

Abstract

Flow cytometric analysis employing monoclonal antibodies to the Tn antigen and glycophorin A was used to characterize the erythrocyte populations present in blood samples from individuals with Tn syndrome. Four monoclonal antibodies specific for the Tn antigen, GalNAc monosaccharide, on human erythrocytes were obtained from a fusion of splenocytes from a Biozzi mouse immunized with red cells from a Tn individual. These monoclonal antibodies specifically recognize GalNAc monosaccharide sites located on the erythrocyte cell surface sialoglycoproteins, glycophorin A and glycophorin B, and do not bind to fixed normal red cells presenting the Neu-NAcα2-3Galβ1-3(NeuNAcα2-6)GalNAcα1-O-Ser(Thr) tetrasaccharide or to fixed neuraminidase-digested cells presenting the Gal-GalNAc disaccharide. The percentages of Tn-positive red cells in samples from six unrelated Tn donors ranged from 28 to 99%. Binding of the glycophorin A-specific monoclonal antibodies showed that the erythrocytes composing the Tn-negative fraction presented normal amounts of the M and N epitopes on glycophorin A. The presumed somatic mutational origin of Tn-positive cells was tested in blood samples from five normal donors; three possible Tn cells were observed after analysis of a total of 1.1 X 107 erythrocytes, suggesting that the frequency of such cells in normal individuals is > 1 X 10−6.

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