Petty H R, Hermann W, Dereski W, Frey T, McConnell H M
J Cell Sci. 1984 Dec;72:1-13. doi: 10.1242/jcs.72.1.1.
Natural killer (NK) cells have been obtained from mouse spleens and grown in vitro. These cells: retained cytotoxicity against YAC targets; and were homogeneous as judged by morphology and surface markers. The alpha-naphthol acetate esterases (ANAE) and diisopropyl-fluorophosphate (DFP)-binding proteins of NK cells, YAC cells and NK-YAC conjugates have been examined. Ultrastructural cytochemistry indicates that NK cells have two types of ANAE: an enzyme primarily associated with the granule externum and an activatable ANAE associated with the granule internum. Both of these activities are inhibited by DFP. The activatable ANAE appears during incubation of NK cells with YAC cells. DFP-binding proteins were examined by sodium dodecyl sulphate/polyacrylamide gel electrophoresis and autofluorography. NK cells and YAC cells have major DFP-binding proteins of 35 X 10(3) and 20 X 10(3) Mr, respectively. NK-YAC conjugates had a new band at 55 X 10(3) Mr. This new protein may be identical to the activatable ANAE described above. Our studies provide evidence for an NK cell esterase that becomes activated when incubated in the presence of tumour cells. This esterase may be related to the cytolytic mechanism.
自然杀伤（NK）细胞已从小鼠脾脏中获取并在体外培养。这些细胞：对YAC靶细胞保持细胞毒性；并且根据形态学和表面标志物判断是均一的。已检测了NK细胞、YAC细胞和NK - YAC结合物的α - 萘酚醋酸酯酶（ANAE）和二异丙基氟磷酸（DFP）结合蛋白。超微结构细胞化学表明NK细胞有两种类型的ANAE：一种主要与颗粒外部相关的酶和一种与颗粒内部相关的可激活的ANAE。这两种活性均被DFP抑制。可激活的ANAE在NK细胞与YAC细胞孵育期间出现。通过十二烷基硫酸钠/聚丙烯酰胺凝胶电泳和自动放射自显影检测DFP结合蛋白。NK细胞和YAC细胞分别有分子量为35×10³和20×10³的主要DFP结合蛋白。NK - YAC结合物在分子量55×10³处有一条新带。这种新蛋白质可能与上述可激活的ANAE相同。我们的研究为一种在肿瘤细胞存在下孵育时被激活的NK细胞酯酶提供了证据。这种酯酶可能与细胞溶解机制有关。
