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荧光素标记的凝集素与小鼠胸腺细胞的差异结合:流式微量荧光测定法揭示的亚群

Differential binding of fluorescein-labeled lectins to mouse thymocytes: subsets revealed by flow microfluorometry.

作者信息

Fowlkes B J, Waxdal M J, Sharrow S O, Thomas C A, Asofsky R, Mathieson B J

出版信息

J Immunol. 1980 Aug;125(2):623-30.

PMID:6967084
Abstract

Fluorescein-labeled lectins bound to mouse thymocytes were analyzed by flow microfluorometry. This technique has identified several lectins that bind differentially to thymocyte subsets. The most complex fluorescence distributions were obtained using lectins with nominal specificities for galactose or N-acetylglucosamine. Inhibition of binding by sugars confirmed that the fluoresceinated lectins were bound to cells at their carbohydrate binding site. Simultaneous analyses of lectin fluorescence and forward light scatter intensity showed that cell subpopulations of different sizes can exhibit marked differences in the level of binding such that the amount of lectin bound per cell is often independent of cell size. A minor population of dull or unstained cells, delineated by several of these lectins, correlates with the subpopulation of medium-sized thymocytes resistant to in vivo cortisone treatment.

摘要

通过流式微量荧光测定法分析了与小鼠胸腺细胞结合的荧光素标记凝集素。该技术已鉴定出几种与胸腺细胞亚群有不同结合方式的凝集素。使用对半乳糖或N-乙酰葡糖胺具有名义特异性的凝集素可获得最复杂的荧光分布。糖对结合的抑制作用证实了荧光素化凝集素在其碳水化合物结合位点与细胞结合。对凝集素荧光和前向光散射强度的同步分析表明,不同大小的细胞亚群在结合水平上可表现出显著差异,以至于每个细胞结合的凝集素量通常与细胞大小无关。由其中几种凝集素勾勒出的一小部分暗淡或未染色的细胞,与对体内可的松治疗有抗性的中等大小胸腺细胞亚群相关。

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