Crouch S P, Kozlowski R, Slater K J, Fletcher J
Medical Research Centre, City Hospital, Nottingham, UK.
J Immunol Methods. 1993 Mar 15;160(1):81-8. doi: 10.1016/0022-1759(93)90011-u.
Adenosine triphosphate (ATP) bioluminescence was used to determine whether there was a linear relationship between cultured cell number and measured luminescence using the luciferin-luciferase reaction. In all the cells tested including peripheral blood mononuclear cells (MNC), MOLT-4, HL-60, TF-1, NFS-60 and L-929 cell lines there was a significant correlation as determined by Spearman's rank correlation coefficient (p > 0.00001). These observations were then used to determine whether ATP bioluminescence could be used as a suitable substitute for tritiated thymidine uptake as a measure of cell proliferation. The cell lines MOLT-4, HL-60, TF-1 and NFS-60 showed a strong correlation between thymidine uptake and ATP bioluminescence (p > 0.00001 for all cell types). Additionally the ATP method could detect the cytokine dependent proliferation on TF-1 and NFS-60 cells by granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (G-CSF) respectively. The tumour necrosis factor alpha (TNF)-induced cytotoxic effect on L-929 cells could also be accurately detected using this method. It would therefore appear to be possible to use ATP bioluminescence in the detection of cytokine activity in a number of different bioassays.
利用三磷酸腺苷（ATP）生物发光法，通过荧光素-荧光素酶反应来确定培养的细胞数量与所测发光值之间是否存在线性关系。在所测试的所有细胞中，包括外周血单个核细胞（MNC）、MOLT-4、HL-60、TF-1、NFS-60和L-929细胞系，经斯皮尔曼等级相关系数测定，均存在显著相关性（p>0.00001）。然后利用这些观察结果来确定ATP生物发光法是否可以作为氚标记胸腺嘧啶核苷摄取法的合适替代方法，用以测量细胞增殖。MOLT-4、HL-60、TF-1和NFS-60细胞系显示出胸腺嘧啶核苷摄取与ATP生物发光之间存在强相关性（所有细胞类型的p>0.00001）。此外，ATP方法能够分别检测粒细胞-巨噬细胞集落刺激因子（GM-CSF）对TF-1细胞以及粒细胞集落刺激因子（G-CSF）对NFS-60细胞的细胞因子依赖性增殖。使用该方法还能准确检测肿瘤坏死因子α（TNF）对L-929细胞的细胞毒性作用。因此，在许多不同的生物测定中，似乎可以使用ATP生物发光法来检测细胞因子活性。
