Ezaki T, Christensen N D, Skinner M A, Marbrook J
J Immunol Methods. 1984 Feb 10;66(2):357-68. doi: 10.1016/0022-1759(84)90349-1.
The use of the original haemolytic plaque reduction technique to measure cytotoxic T lymphocytes (CTL) has been developed further as a rapid screening assay, particularly suitable for limiting dilution analyses. Using hybridoma cells as targets, the cytotoxicity has been measured by the loss of haemolytic plaque formation and by the reduction of the amount of haemolytic monoclonal antibody secreted from viable target cells into the assay supernatants. The assessment of large numbers of cytotoxic samples has been greatly facilitated by quantitating the amount of haemoglobin released in the assay with an automated microELISA multiscanner and by scoring visually using a modification of the spot test. Using these new techniques, relatively high frequency estimates of cytotoxic cell precursors in an allogeneic response (1 in 462 spleen cells) and an anti-fluorescein response (1 in 3970 spleen cells) were obtained.
最初用于测量细胞毒性T淋巴细胞（CTL）的溶血空斑减少技术已进一步发展成为一种快速筛选检测方法，特别适用于有限稀释分析。以杂交瘤细胞作为靶细胞，通过溶血空斑形成的丧失以及从存活靶细胞分泌到检测上清液中的溶血单克隆抗体量的减少来测量细胞毒性。通过使用自动微量酶联免疫吸附测定多扫描仪对检测中释放的血红蛋白量进行定量，并使用斑点试验的改良方法进行目视评分，极大地促进了对大量细胞毒性样品的评估。使用这些新技术，在同种异体反应（每462个脾细胞中有1个）和抗荧光素反应（每3970个脾细胞中有1个）中获得了细胞毒性细胞前体的相对高频估计值。
