Flow cytometric analysis of erythrocyte populations in Tn syndrome blood using monoclonal antibodies to glycophorin A and the Tn antigen.
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, Gal-NAc 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-NAc alpha 2-3Gal beta 1-3(NeuNAc alpha 2-6)GalNAc alpha 1-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 10(7) erythrocytes, suggesting that the frequency of such cells in normal individuals is less than 1 x 10(-6).
Bigbee
W L
WL
Sciences Division, Lawrence Livermore National Laboratory, University of California, Livermore 94550.
Langlois
R G
RG
Stanker
L H
LH
Vanderlaan
M
M
Jensen
R H
RH
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm / Q000276:immunology*; D015295:Antigens, Tumor-Associated, Carbohydrate; D015415:Biomarkers; D001789:Blood Group Antigens / Q000276:immunology; D002469:Cell Separation; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis; D004912:Erythrocytes / Q000166:cytology* / Q000276:immunology; D005434:Flow Cytometry; D006021:Glycophorins / Q000276:immunology*; D006386:Hemagglutination Tests; D006801:Humans; D008951:MNSs Blood-Group System; D009154:Mutation; D012795:Sialoglycoproteins / Q000276:immunology*; D013577:Syndrome
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D015295:Antigens, Tumor-Associated, Carbohydrate; D015415:Biomarkers; D001789:Blood Group Antigens; D000939:Epitopes; D006021:Glycophorins; D008951:MNSs Blood-Group System; D012795:Sialoglycoproteins; C040286:Tn antigen