Adachi Reiko, Takeuchi Kosei, Suzuki Kazuhiro
National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan.
J Biol Chem. 2002 Nov 22;277(47):45566-71. doi: 10.1074/jbc.M207419200. Epub 2002 Sep 23.
Phagocytes play a central role in the host defense system, and the relationship between the mechanism of their activation and cytoskeletal reorganization has been studied. We have previously reported a possible involvement of cofilin, an actin-binding protein, in phagocyte functions through its phosphorylation/dephosphorylation and translocation to the plasma membrane regions. In this work, we have obtained a new line of evidence showing an important role of cofilin in phagocyte functions using the mouse macrophage cell line J774.1 and an antisense oligonucleotide to cofilin. Upon stimulation with opsonized zymosan (OZ), cofilin was phosphorylated, and it accumulated around phagocytic vesicles. As the antisense oligonucleotide to cofilin, a 20-mer S-oligo corresponding to the sequence including the AUG translational initiation site was found to be effective. In the cells treated with the antisense oligonucleotide, the amount of cofilin was less than 30% of that in the control cells, and the level of F-actin was two or three times higher than that in the control cells before and throughout the cell activation. In the antisense oligonucleotide-treated cells, OZ-triggered superoxide production was three times faster than that in the control cells. Furthermore, phagocytosis of OZ was enhanced by the antisense. These results show that cofilin plays an essential role in the control of phagocyte function through regulation of actin filament dynamics.
吞噬细胞在宿主防御系统中发挥着核心作用,并且其激活机制与细胞骨架重组之间的关系已得到研究。我们之前曾报道,肌动蛋白结合蛋白丝切蛋白可能通过其磷酸化/去磷酸化以及向质膜区域的转位参与吞噬细胞功能。在这项研究中,我们利用小鼠巨噬细胞系J774.1和针对丝切蛋白的反义寡核苷酸获得了新的证据,表明丝切蛋白在吞噬细胞功能中具有重要作用。在用调理酵母聚糖(OZ)刺激后,丝切蛋白发生磷酸化,并在吞噬小泡周围积累。作为针对丝切蛋白的反义寡核苷酸,一种与包含AUG翻译起始位点的序列相对应的20聚体硫代寡核苷酸被发现是有效的。在用反义寡核苷酸处理的细胞中,丝切蛋白的量不到对照细胞的30%,并且在细胞激活之前及整个过程中,F-肌动蛋白水平比对照细胞高两到三倍。在用反义寡核苷酸处理的细胞中,OZ触发的超氧化物产生比对照细胞快三倍。此外,反义寡核苷酸增强了对OZ的吞噬作用。这些结果表明,丝切蛋白通过调节肌动蛋白丝动力学在控制吞噬细胞功能中发挥着重要作用。