Fuller S, von Bonsdorff C H, Simons K
Cell. 1984 Aug;38(1):65-77. doi: 10.1016/0092-8674(84)90527-0.
We have used Madin-Darby canine kidney (MDCK) cells grown on nitrocellulose filters to study the polarity of virus infection and maturation. The cells form epithelia-like monolayers, which display high (greater than 1000 omega cm2) electrical resistance and a cuboidal morphology. Vesicular stomatitis virus (VSV) was found to infect the monolayer at least 100 times more efficiently when applied through the filter to the basolateral surface than when applied to the apical surface. The avian influenza, fowl plague virus (FPV), infected the monolayer through either the apical or basolateral surface. The polarity of virus budding was evaluated by harvesting virus from the two sides of the monolayer. More than 99% of released influenza hemagglutinin titre was found on the apical side of the filter, while more than 98% of budded VSV was found on the basal side. This polarity of budding was retained through 10 hr of viral infection, as was the polarity of surface expression of viral envelope proteins revealed by immunofluorescence. The strong preference of VSV for basolateral maturation is paralleled by an equally strong preference for infection through the basolateral membrane of this polar epithelial cell.
我们利用在硝酸纤维素滤膜上生长的犬肾传代细胞(MDCK)来研究病毒感染和成熟的极性。这些细胞形成上皮样单层,具有高(大于1000Ω·cm²)电阻和立方形形态。发现水泡性口炎病毒(VSV)通过滤膜施加于基底外侧表面时感染单层细胞的效率比施加于顶端表面时至少高100倍。禽流感病毒、鸡瘟病毒(FPV)可通过顶端或基底外侧表面感染单层细胞。通过从单层细胞的两侧收获病毒来评估病毒出芽的极性。在滤膜顶端一侧发现超过99%的释放型流感血凝素滴度,而在基底一侧发现超过98%的出芽VSV。这种出芽极性在病毒感染10小时内得以保持,通过免疫荧光显示的病毒包膜蛋白表面表达极性也是如此。VSV对基底外侧成熟的强烈偏好与对通过这种极性上皮细胞的基底外侧膜进行感染的同样强烈偏好相平行。