Kaplan J H, Uzgiris E E
J Immunol Methods. 1975 Jul;7(4):337-46. doi: 10.1016/0022-1759(75)90042-3.
The changes in electrophoretic mobility and isoelectric point produced by incubating human peripheral blood lymphocytes with phytohemagglutinin (PHA) and concanavalin A (Con A) have been characterized by laser Doppler spectroscopy. The results extend and partially confirm older observations made by classical procedures. Incubation with either agent for 90 min at 37 degrees C resulted in stable and reproducible decreases in electrophoretic mobility, and increases in the isoelectric point. The incubation conditions used are known to permit primary attachment of the phytomitogen, capping and endocytosis; nevertheless, at least in the case of Con A, washing the cells with a specific inhibitor for Con A binding, methyl-alpha-D-glucoside (MAG), resulted in complete reversal of the electrokinetic changes, showing that the underlying changes in cell surface constitution detected under these conditions are solely due to reversibly bound Con A. The results suggest that laser Doppler spectroscopic changes could provide a direct assay for specific binding to immunocompetent cell surfaces.
通过激光多普勒光谱法对用植物血凝素（PHA）和刀豆球蛋白A（Con A）孵育人外周血淋巴细胞所产生的电泳迁移率和等电点的变化进行了表征。结果扩展并部分证实了通过经典方法所做的早期观察。在37℃下用任一种试剂孵育90分钟导致电泳迁移率稳定且可重复地降低，以及等电点升高。已知所使用的孵育条件允许植物促细胞分裂剂的初始附着、帽化和内吞作用；然而，至少在Con A的情况下，用Con A结合的特异性抑制剂α-D-甲基葡萄糖苷（MAG）洗涤细胞导致电动变化完全逆转，表明在这些条件下检测到的细胞表面组成的潜在变化完全是由于可逆结合的Con A所致。结果表明，激光多普勒光谱变化可为与免疫活性细胞表面的特异性结合提供直接测定方法。
