de Matos A P, Carvalho Z G
Pathology Department, Curry Cabral Hospital, Lisbon, Portugal.
Biol Cell. 1993;78(3):229-34. doi: 10.1016/0248-4900(93)90134-z.
The role of microtubules in intracellular transport of African swine fever virus (ASFV) and virus-induced inclusions was studied by immunofluorescence using anti-ASFV and anti-tubulin antibodies, by electron microscopy of infected Vero cells and by in vitro binding of virions to purified microtubules. MTC, a reversible colchicine analogue, was used to depolymerize microtubules. In cells treated with MTC multiple large inclusions containing ASFV antigens and particles were observed in the cytoplasm. Removal of the drug lead to migration and fusion of the inclusions at a perinuclear location. To study the effect of microtubule repolymerization on virus particle distribution, the particles were counted in thin sections of MTC treated cells and at different times after removal of the drug. In cells treated with MTC 6.8% and 3.6% of the virus particles were found respectively in the cytoplasm and at the cell membrane while 38% of the particles were located around the virosome. With reversal of the drug effect the number of virus particles around the virosomes progressively decreased to 10% at 2 h while the number of particles in the cytoplasm and at the cell membrane increased. At 2 h after removal of the drug 33.5% of the particles were found budding from the cell membrane. Virus particles were found closely associated with microtubules in cytoskeletons obtained by Triton X-100 extraction of taxol treated cells. The association of virus particles with microtubules was also observed in vitro using purified microtubules and virus particles.(ABSTRACT TRUNCATED AT 250 WORDS)
通过使用抗非洲猪瘟病毒（ASFV）抗体和抗微管蛋白抗体进行免疫荧光检测、对感染的非洲绿猴肾（Vero）细胞进行电子显微镜观察以及使病毒粒子与纯化的微管进行体外结合，研究了微管在ASFV细胞内运输及病毒诱导包涵体形成中的作用。使用秋水仙碱类似物MTC使微管解聚。在用MTC处理的细胞中，在细胞质中观察到多个含有ASFV抗原和粒子的大包涵体。去除药物后，包涵体迁移并在核周位置融合。为研究微管重新聚合对病毒粒子分布的影响，对MTC处理的细胞薄片以及去除药物后的不同时间点的病毒粒子进行计数。在用MTC处理的细胞中，分别有6.8%和3.6%的病毒粒子位于细胞质和细胞膜，而38%的粒子位于病毒体周围。随着药物作用的逆转，病毒体周围的病毒粒子数量在2小时时逐渐减少至10%，而细胞质和细胞膜中的粒子数量增加。去除药物2小时后，发现33.5%的粒子从细胞膜出芽。在通过用Triton X-100提取紫杉醇处理的细胞获得的细胞骨架中，发现病毒粒子与微管紧密相关。使用纯化的微管和病毒粒子在体外也观察到了病毒粒子与微管的结合。（摘要截短于250字）
