Volgmann T, Klein-Struckmeier A, Mohr H
Blood Transfusion Service of Lower Saxony, Department of Research and Development, Springe, F.R.G.
J Immunol Methods. 1989 Apr 21;119(1):45-51. doi: 10.1016/0022-1759(89)90379-7.
A fluorescence assay for the quantitation of tumor cell lysis by activated and non-activated killer (LAK) cells is described. The target cells are labelled with a europium chelate (Eu-diethylenetriaminopentaacetate) and after cytolysis caused by the LAK cells the Eu3+ complex is released into the culture supernatant. The addition of beta-naphthoyltrifluoroacetone to culture supernatant aliquots leads to the formation of a highly fluorescent chelate which can be measured with a time-resolved fluorometer. The influence of various assay parameters has been evaluated including incubation time, effector-to-target cell ratio, the target cell line and different concentrations of interleukin-2 during cell culture. The optimized time-resolved fluorometric assay was found to be as simple and sensitive as the commonly used cytotoxicity assay in which the release of 51Cr from the labelled target cells is measured. In addition the assay is much faster and safer since the label is not radioactive.
本文描述了一种用于定量活化和未活化杀伤（LAK）细胞对肿瘤细胞裂解作用的荧光测定法。靶细胞用铕螯合物（铕-二乙烯三胺五乙酸）标记，在LAK细胞引起细胞溶解后，Eu3+复合物释放到培养上清液中。向上清液等分试样中加入β-萘甲酰三氟丙酮会导致形成一种高荧光螯合物，可用时间分辨荧光计进行测量。评估了各种测定参数的影响，包括孵育时间、效应细胞与靶细胞比例、靶细胞系以及细胞培养过程中不同浓度的白细胞介素-2。结果发现，优化后的时间分辨荧光测定法与常用的细胞毒性测定法一样简单灵敏，后者是通过测量标记靶细胞中51Cr的释放来进行的。此外，该测定法更快且更安全，因为标记物不是放射性的。
