Bigbee W L, Langlois R G, Stanker L H, Vanderlaan M, Jensen R H
Sciences Division, Lawrence Livermore National Laboratory, University of California, Livermore 94550.
Cytometry. 1990;11(2):261-71. doi: 10.1002/cyto.990110207.
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).
采用针对Tn抗原和血型糖蛋白A的单克隆抗体进行流式细胞术分析，以表征Tn综合征患者血样中存在的红细胞群体。从用Tn个体的红细胞免疫的Biozzi小鼠的脾细胞融合物中获得了四种针对人红细胞上Tn抗原（Gal-NAc单糖）的单克隆抗体。这些单克隆抗体特异性识别位于红细胞表面唾液糖蛋白、血型糖蛋白A和血型糖蛋白B上的GalNAc单糖位点，并且不与呈现Neu-NAcα2-3Galβ1-3（NeuNAcα2-6）GalNAcα1-O-Ser（Thr）四糖的固定正常红细胞结合，也不与呈现Gal-GalNAc二糖的固定神经氨酸酶消化细胞结合。来自六个不相关Tn供体的样本中Tn阳性红细胞的百分比范围为28%至99%。血型糖蛋白A特异性单克隆抗体的结合表明，构成Tn阴性部分的红细胞在血型糖蛋白A上呈现正常量的M和N表位。在来自五个正常供体的血样中测试了Tn阳性细胞推测的体细胞突变起源；在总共分析了1.1×10⁷个红细胞后观察到三个可能的Tn细胞，这表明正常个体中此类细胞的频率小于1×10⁻⁶。
