Eskinazi D P, Hasson J N, Kebo D K
Immunol Lett. 1982 Sep;5(3):161-6. doi: 10.1016/0165-2478(82)90102-x.
This report describes the use of fibronectin-coated polyvinyl chloride (PVC) plates as a time-saving modification in an assay measuring antibody-binding to live adherent tumor cells. Three cell lines (A-431, Colo 16 and UCLA-SO-P3) derived from human squamous cell carcinomas (SCC) and forming monolayers in cultures were plated onto flexible PVC microtest plates rather than the commonly used rigid polystyrene plates. In PVC plates, two of the three cell lines (A-431 and Colo 16) grew as foci of clumped cells instead of monolayers. Coating of the plates with plasma fibronectin restored the monolayer morphology. [125I]Staphylococcal protein A radioimmunoassays measuring the binding of a monoclonal anti-beta 2 microglobulin antibody to the cells were slightly, but consistently, more sensitive in coated PVC plates than in polystyrene plates. In contrast, the sensitivity of the radioimmunoassay was remarkably constant when the assay was performed on the third cell line (UCLA-SO-P3), which formed monolayers in any of the conditions tested. Preliminary experiments suggested that the inability to form monolayers on uncoated PVC plates correlates with the amount of fibronectin associated with the cell surface.
本报告描述了使用纤连蛋白包被的聚氯乙烯（PVC）板作为一种节省时间的改进方法，用于检测抗体与活的贴壁肿瘤细胞的结合。将源自人鳞状细胞癌（SCC）并在培养中形成单层的三种细胞系（A-431、Colo 16和UCLA-SO-P3）接种到柔性PVC微量测试板上，而不是常用的刚性聚苯乙烯板上。在PVC板中，三种细胞系中的两种（A-431和Colo 16）以细胞团块的形式生长，而不是形成单层。用血浆纤连蛋白包被板可恢复单层形态。[125I]葡萄球菌蛋白A放射免疫分析检测单克隆抗β2微球蛋白抗体与细胞的结合，在包被的PVC板中比在聚苯乙烯板中稍微但始终更敏感。相比之下，当对第三种细胞系（UCLA-SO-P3）进行该分析时，放射免疫分析的灵敏度非常恒定，该细胞系在任何测试条件下均形成单层。初步实验表明，在未包被的PVC板上无法形成单层与细胞表面相关的纤连蛋白量有关。
