Skosey J L, Damgaard E, Chow D, Sorensen L B
J Cell Biol. 1974 Sep;62(3):625-34. doi: 10.1083/jcb.62.3.625.
During the process of phagocytosis, polymorphonuclear leukocytes (PMN) release lysosomal enzymes into the extracellular medium. When the antibiotic cytochalasin B (CB) is present in the incubation medium along with phagocytable particles, enhanced recovery of enzyme activities from the incubation medium has been observed. These findings have led to the interpretation that CB enhances lysosomal enzyme release. Our results contradict this interpretation. The lysosomal enzymes acid phosphatase and beta-galactosidase are unstable after they are released from cells. During the first 5-15 min of phagocytosis, significant amounts of both acid phosphatase and beta-galactosidase can be recovered from the extracellular medium. After this, the recovery of enzyme from the medium declines, presumably because the rate of loss of lysosomal enzyme activity exceeds the rate of release at later time periods. In the presence of CB, the appearance of lysosomal enzymes in the extracellular medium of cells exposed to zymosan is retarded for 5-10 min, after which it begins and then continues for approximately 20 min. At the end of a 30-min incubation period, therefore, in the absence of CB, extracellular levels of lysosomal enzymes (especially those which are unstable) are declining toward low levels while, in the presence of CB, extracellular enzyme levels are continuing to rise. We also measured the lysosomal enzyme remaining within cells after exposure to zymosan. CB retarded the disappearance of enzyme from cells and resulted in significantly less total cell enzyme loss. Thus, in the presence of CB, a greater proportion of the lysosomal enzyme lost from cells is recovered in the extracellular medium. In contrast to the previous conclusions that CB enhances lysosomal enzyme release, our results indicate that CB delays and decreases the zymosan-stimulated release of lysosomal enzymes from PMN. Since CB inhibits phagocytosis by PMN, our results indicate that the antibiotic modifies the mechanism of release of lysosomal enzymes, resulting in zymosan stimulation of their release independently of phagocytosis.
在吞噬过程中,多形核白细胞(PMN)会将溶酶体酶释放到细胞外介质中。当抗生素细胞松弛素B(CB)与可吞噬颗粒一起存在于孵育介质中时,已观察到从孵育介质中回收的酶活性增强。这些发现导致了CB增强溶酶体酶释放的解释。我们的结果与这一解释相矛盾。溶酶体酶酸性磷酸酶和β-半乳糖苷酶从细胞释放后不稳定。在吞噬的最初5 - 15分钟内,细胞外介质中可回收大量的酸性磷酸酶和β-半乳糖苷酶。在此之后,从介质中回收的酶量下降,推测是因为溶酶体酶活性丧失的速率超过了后期释放的速率。在CB存在的情况下,暴露于酵母聚糖的细胞的细胞外介质中溶酶体酶的出现延迟5 - 10分钟,之后开始并持续约20分钟。因此,在30分钟的孵育期结束时,在没有CB的情况下,溶酶体酶的细胞外水平(尤其是那些不稳定的酶)正在下降至低水平,而在有CB的情况下,细胞外酶水平仍在继续上升。我们还测量了暴露于酵母聚糖后细胞内剩余的溶酶体酶。CB延缓了酶从细胞中的消失,并导致细胞总酶损失显著减少。因此,在CB存在的情况下,从细胞中损失的溶酶体酶有更大比例在细胞外介质中被回收。与之前关于CB增强溶酶体酶释放的结论相反,我们的结果表明CB延迟并减少了酵母聚糖刺激的PMN溶酶体酶释放。由于CB抑制PMN的吞噬作用,我们的结果表明该抗生素改变了溶酶体酶的释放机制,导致酵母聚糖独立于吞噬作用刺激其释放。