Peddie C M, Smith V J
School of Biological and Medical Sciences, Gatty Marine Laboratory, St. Andrews University, Fife, Scotland.
J Exp Zool. 1993 Dec 15;267(6):616-23. doi: 10.1002/jez.1402670609.
Blood cell-mediated cytotoxic activity against mammalian target cells by the hemocytes of the solitary ascidian Ciona intestinalis was investigated in vitro by fluorochromasia. Salt-conditioned target cells were labeled with carboxyfluorescein diacetate and challenged with mixed and separated hemocytes. The assay provided optimal conditions for the functioning of the effector hemocytes while maintaining low background leakage from the target cells. Comparison of different hemocyte populations, separated by density gradient centrifugation, revealed that only cell bands containing the phagocytic and nonphagocytic amoebocytes exhibited cytotoxicity. Experiments to characterize cytolysis demonstrated that activity increased with the effector to target cell ratio, occurred within 15 min, and was maximal at an incubation temperature of 20 degrees C. Both human (K562) and mouse [YAC-1, P815, WEHI (3B) and L929] target cell lines were killed by the ascidian effector hemocytes. This paper demonstrates a population of nonspecific cytotoxic effector cells in the blood of C. intestinalis that are able to spontaneously kill a range of mammalian targets in vitro.
通过荧光染色法在体外研究了单体海鞘玻璃海鞘血细胞对哺乳动物靶细胞的血细胞介导的细胞毒性活性。用羧基荧光素二乙酸酯标记盐处理过的靶细胞，并用混合和分离的血细胞进行攻击。该测定为效应血细胞的功能提供了最佳条件，同时保持靶细胞的低背景泄漏。通过密度梯度离心分离不同血细胞群体的比较表明，只有含有吞噬性和非吞噬性变形细胞的细胞带表现出细胞毒性。表征细胞溶解的实验表明，活性随效应细胞与靶细胞的比例增加而增加，在15分钟内发生，并且在20摄氏度的孵育温度下最大。人（K562）和小鼠[YAC-1、P815、WEHI（3B）和L929]靶细胞系均被海鞘效应血细胞杀死。本文证明了玻璃海鞘血液中存在一群非特异性细胞毒性效应细胞，它们能够在体外自发杀死一系列哺乳动物靶标。
