Lewis P, Fu Y, Lentz T L
Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06520-8002, USA.
Exp Neurol. 1998 Sep;153(1):65-73. doi: 10.1006/exnr.1998.6879.
Early events in rabies virus entry into cultured IMR-32 human neuroblastoma cells were investigated. After adsorption of rabies virus to the cell surface in the cold and warming to 37 degrees C in the presence of tracers for early endosomes, rabies virus and tracers were localized by immunofluorescence microscopy. After 5 min, rabies virus colocalized with Lucifer Yellow, Texas Red-dextran, rhodamine-wheat germ agglutinin, and transferrin receptor in puncta in the cell body, neurites, and nerve terminals. Rabies virus did not colocalize with lysosomal glycoprotein. An acidotropic probe revealed that some of the virus-containing puncta were acidified. Rabies virus also colocalized with synapsin I, a synaptic vesicle marker, in swellings along processes, indicating some virus enters nerve terminals. Electron microscopy revealed the presence of rabies virus within irregular membrane compartments located near the cell surface in the cell body and neurites. The membrane of the virus particle was often continuous with that of the vacuole. It is concluded that rabies virus enters IMR-32 neuroblastoma cells by adsorptive endocytosis and that, shortly after entry, rabies virus is located within and fuses with acidic endosomes.
研究了狂犬病毒进入培养的IMR-32人神经母细胞瘤细胞的早期事件。在低温条件下狂犬病毒吸附到细胞表面并在存在早期内体示踪剂的情况下升温至37摄氏度后，通过免疫荧光显微镜对狂犬病毒和示踪剂进行定位。5分钟后，狂犬病毒与荧光素黄、德克萨斯红葡聚糖、罗丹明-小麦胚凝集素和转铁蛋白受体在细胞体、神经突和神经末梢的点状结构中共定位。狂犬病毒与溶酶体糖蛋白不共定位。一种嗜酸性探针显示一些含有病毒的点状结构被酸化。狂犬病毒还与突触小泡蛋白I（一种突触小泡标记物）在沿突起的肿胀部位共定位，表明一些病毒进入神经末梢。电子显微镜显示在细胞体和神经突中靠近细胞表面的不规则膜性区室中存在狂犬病毒。病毒颗粒的膜通常与液泡的膜连续。得出的结论是，狂犬病毒通过吸附性内吞作用进入IMR-32神经母细胞瘤细胞，并且在进入后不久，狂犬病毒位于酸性内体中并与之融合。
