THE USE OF NONRADIOACTIVE CHROMIUM AS AN ALTERNATIVE TO CR-51 IN NK ASSAY
THE USE OF NONRADIOACTIVE CHROMIUM AS AN ALTERNATIVE TO CR-51 IN NK ASSAY
复制标题
DOI：
10.1016/0022-1759(95)00140-6
复制
发表时间：
1995-10-12
影响因子：
2.2
通讯作者：
COSSARIZZA, A
中科院分区：
文献类型：
--
作者：
BORELLA, P;BARGELLINI, A;COSSARIZZA, A
提出了一种基于使用“冷”、非放射性铬和通过石墨炉原子吸收光谱法（FAAS）测定金属释放的测量靶细胞溶解的新方法。自然杀伤细胞（NK）的测定进行标记的靶细胞与铬作为Na 2CrO 4，并与那些通过常规的过夜标记与Cr-51的目标杀死相同的效应得到的结果进行了比较。评价了来自健康受试者的外周血淋巴细胞的细胞毒性能力，并且用两种方法测量的NK活性在每个测试的效应细胞与靶细胞比率（在100：1和1：1之间）下显示出良好的一致性，具有高且显著的相关系数（r = 0.931，p < 0.0001）。标记靶细胞的适当铬浓度的选择考虑了我们的仪器的灵敏度和金属的可能毒性作用。一项关于铬对NK试验中通常用作靶细胞的细胞系（K562）的影响的研究表明，铬可能对细胞功能产生有害影响，在使用高于15 μ mol/l（780 μ g/l）的铬水平孵育24小时后，大量细胞线粒体去极化，DNA合成减少。这里提出的方法具有许多优点，包括使用非放射性示踪剂，有限的成本，高灵敏度和再现性，以及储存样品的可能性。此外，该技术使用已知无毒的固定Cr浓度。
A novel method to measure target cell cytolysis based on the use of 'cold', non-radioactive chromium and on the determination of metal release by graphite furnace atomic absorption spectroscopy (FAAS) is proposed. Natural killer (NK) assays were performed by labelling target cells with chromium as Na2CrO4, and results were compared with those obtained by conventional overnight labelling with Cr-51 of targets killed by the same effecters. The cytotoxic capacity of peripheral blood lymphocytes from healthy subjects was evaluated, and NK activity measured with both methods showed a good agreement at each of the tested effector to target cell ratios (between 100:1 and 1:1), with a high and significant coefficient of correlation (r = 0.931, p < 0.0001). The selection of the appropriate Cr concentrations for labelling target cells took into account both the sensitivity of our instrumentation and the possible toxic effects of the metal. A study of the effects of Cr on the cell line (K562) which is usually employed as a target in NK tests showed that Cr could have a detrimental effect on cellular function, with significant numbers of cells with depolarised mitochondria and reduced DNA synthesis after 24 h incubation using Cr levels higher than 15 mu mol/l (780 mu g/l). The method proposed here has a number of advantages, including the use of a non-radioactive tracer, limited costs, high sensitivity and reproducibility, and the possibility of storing samples. In addition, the technique uses a fixed Cr concentration which is known to be non toxic.