Keilholz U, Dummer R, Welters H, Brado B, Galm F, Matheiowetz P, Hunstein W
Medizinische Klinik und Poliklinik V, University of Heidelberg, FRG.
Scand J Clin Lab Invest. 1990 Dec;50(8):879-84. doi: 10.3109/00365519009104956.
Since adoptive immunotherapy using lymphokine activated killer (LAK) cells and interleukin-2 (IL-2) has been introduced into clinical medicine, there has been a growing interest in cytotoxicity assays. The standard 51chromium release assay has certain disadvantages, in particular limited sensitivity, because of a high, nonspecific background release. We examined the conditions under which tritiated thymidine, which has been used to assess slow macrophage mediated cytolysis, can be utilized to assess the rapid cytotoxic activity of unstimulated PBL and LAK cells. The optimal assay duration for the 3H-thymidine (3H-TdR) release assay is 24 h. Under standard conditions actively proliferating effector cells do incorporate some of the 3H-TdR released by the target cells during this time, leading to false low results. This problem can be abolished by the addition of excessive amounts of cold TdR to the assay medium. We found a good correlation of the results of the TdR release assay and the Cr release assay. Using the very sensitive TdR release assay, unexpected significant cytolysis of the so-called 'NK resistant' Daudi cells by unstimulated PBL is demonstrated. The modified 3H-TdR release assay is well-suited to monitor the immunological effects of immunotherapy, using IL-2 and LAK cells.
自从将使用淋巴因子激活的杀伤细胞（LAK细胞）和白细胞介素-2（IL-2）的过继性免疫疗法引入临床医学以来，人们对细胞毒性测定的兴趣日益浓厚。标准的51铬释放测定法存在某些缺点，特别是由于高非特异性背景释放导致灵敏度有限。我们研究了在何种条件下，已用于评估缓慢巨噬细胞介导的细胞溶解作用的氚化胸腺嘧啶核苷可用于评估未刺激的外周血淋巴细胞（PBL）和LAK细胞的快速细胞毒性活性。3H-胸腺嘧啶核苷（3H-TdR）释放测定的最佳测定持续时间为24小时。在标准条件下，在此期间，活跃增殖的效应细胞确实会摄取靶细胞释放的一些3H-TdR，从而导致错误的低结果。通过向测定培养基中添加过量的冷TdR可以消除这个问题。我们发现TdR释放测定结果与Cr释放测定结果具有良好的相关性。使用非常灵敏的TdR释放测定法，证明未刺激的PBL对所谓的“NK抗性”Daudi细胞有意外的显著细胞溶解作用。改良的3H-TdR释放测定法非常适合监测使用IL-2和LAK细胞的免疫疗法的免疫效果。
