Reimann J, Ehman D, Miller R G
Cytometry. 1984 Mar;5(2):194-203. doi: 10.1002/cyto.990050214.
Murine marrow cells were labeled with a panel of fluorescein-conjugated lectins and analyzed by flow cytometry. All lectins tested [concanavalin A (Con A), phytohemagglutinin (PHA), wheat germ agglutinin (WGA), pokeweed mitogen (PWM), soybean agglutinin (SBA), peanut agglutinin (PNA), lentil lectin (LL)], stained a fraction of marrow cells; the fraction was, over a defined concentration range, linearly related to the logarithm of the lectin concentration. The slope of this titration curve was characteristic of the particular lectin tested but independent of the nominal sugar specificity of the respective lectin. Con A-, PWM-, and PNA-labeled marrow cells showed brightly and dimly stained subsets in relative fluorescence intensity distribution histograms at high or intermediate staining concentrations. These subsets were composed of cells with restricted and characteristic size distributions as determined by forward light scatter. Lectin staining patterns of marrow cells from normal mice were compared with those of athymic nude mice and mice with severe combined immunodeficiency (SCID). Normal and nude mice lacked a subset of small cells, dimly stained by Con A and PWM, but brightly stained by PNA. This subset, uniquely defined by the above lectins, appears to correspond to the lymphopoietic marrow compartment.
用一组荧光素偶联凝集素标记小鼠骨髓细胞,并通过流式细胞术进行分析。所有测试的凝集素[刀豆球蛋白A(Con A)、植物血凝素(PHA)、麦胚凝集素(WGA)、商陆有丝分裂原(PWM)、大豆凝集素(SBA)、花生凝集素(PNA)、扁豆凝集素(LL)]均能使一部分骨髓细胞染色;在规定的浓度范围内,这部分细胞与凝集素浓度的对数呈线性关系。该滴定曲线的斜率是所测试的特定凝集素的特征,但与各凝集素的标称糖特异性无关。在高或中等染色浓度下,Con A、PWM和PNA标记的骨髓细胞在相对荧光强度分布直方图中显示出明亮和暗淡染色的亚群。这些亚群由通过前向光散射确定的具有受限和特征性大小分布的细胞组成。将正常小鼠、无胸腺裸鼠和严重联合免疫缺陷(SCID)小鼠的骨髓细胞凝集素染色模式进行了比较。正常小鼠和裸鼠缺乏一小部分小细胞亚群,这些细胞被Con A和PWM暗淡染色,但被PNA明亮染色。由上述凝集素独特定义的这一亚群似乎对应于淋巴细胞生成的骨髓区室。