Schultze B, Zimmer G, Herrler G
Institut für Virologie, Philipps-Universität Marburg, Germany.
J Gen Virol. 1996 Oct;77 ( Pt 10):2507-14. doi: 10.1099/0022-1317-77-10-2507.
We have analysed the uptake of influenza C virus and bovine coronavirus (BCV) by a polarized epithelial cell line, Madin-Darby canine kidney (MDCK) cells. Both viruses use N-acetyl-9-O-acetylneuraminic acid as a receptor determinant for attachment to cells. Virus binding assays with immobilized proteins indicated that a glycoprotein of 40 kDa is the major surface protein containing the receptor determinant for the two viruses. MDCK cells grown on filters for permeable support were found to have differential sensitivity to infection by these viruses. Both viruses were able to initiate infection via the apical domain of the plasma membrane, but only influenza C virus also accomplished infection via the basolateral plasma membrane. The resistance of MDCK cells to BCV infection from the basal filter chamber was overcome when the cell polarity was abolished by maintaining the cells in calcium-free medium. This finding indicates that the resistance to basolateral infection by BCV is a property of the cell line and not due to a technical problem related to the use of filters. Our results indicate that two viruses which use the same receptor for attachment to cells may differ in their ability to enter polarized cells. The possible involvement of an accessory molecule in the entry of BCV is discussed.
我们分析了极化上皮细胞系——马-达二氏犬肾(MDCK)细胞对丙型流感病毒和牛冠状病毒(BCV)的摄取。两种病毒均利用N-乙酰-9-O-乙酰神经氨酸作为细胞附着的受体决定簇。用固定化蛋白进行的病毒结合试验表明,一种40 kDa的糖蛋白是这两种病毒含受体决定簇的主要表面蛋白。在可渗透支持物滤膜上生长的MDCK细胞对这两种病毒感染具有不同的敏感性。两种病毒都能够通过质膜的顶端结构域引发感染,但只有丙型流感病毒也能通过基底外侧质膜完成感染。当通过在无钙培养基中培养细胞来消除细胞极性时,MDCK细胞对来自基底滤室的BCV感染的抗性就被克服了。这一发现表明,对BCV基底外侧感染的抗性是细胞系的一种特性,而不是由于与使用滤膜相关的技术问题。我们的结果表明,两种利用相同受体附着于细胞的病毒,其进入极化细胞的能力可能不同。文中还讨论了一种辅助分子在BCV进入过程中的可能作用。