Attachment of infectious influenza A viruses of various subtypes to live mammalian and avian cells as measured by flow cytometry
Attachment of infectious influenza A viruses of various subtypes to live mammalian and avian cells as measured by flow cytometry
复制标题
DOI：
10.1016/j.virusres.2007.07.007
复制
发表时间：
2007-11-01
期刊：
影响因子：
5
通讯作者：
Osterhaus, Albert D. M. E.
中科院分区：
文献类型：
--
作者：
Rimmelzwaan, Guus F.;Nieuwkoop, Nella J.;Osterhaus, Albert D. M. E.
目前，人们对流感病毒的细胞嗜性和宿主范围非常感兴趣，尤其是H5N1亚型流感病毒。我们希望开发一种方法，通过血凝素分子与活的哺乳动物和鸟类细胞的相互作用以及这些细胞的后续感染来研究传染性病毒的附着。为此，构建了不同亚型的流感病毒，它们要么携带绿色荧光蛋白(GFP)而不是神经氨酸酶蛋白，要么在感染细胞的细胞质中表达GFP。HA基因来源于流感病毒A/PR/8/34(H1N1)、A/荷兰/178/95(H3N2)和A/越南/1194/04(H5N1)。使用这些病毒对，可以区分病毒复制周期中的附着事件和附着后事件。一般来说，NeuAc(Alpha 2-3)Gal或NeuAc(Alpha 2-6)Gal受体在测试细胞上的表达与所研究的病毒的附着性相对应，这与预测的受体特异性有关。病毒附着并不总是预示着细胞的有效感染。(C)2007 Elsevier B.V.保留所有权利。
At present there is much interest in the cell tropism and host range of influenza viruses, especially those of the H5N1 subtype. We wished to develop a method that would enable investigation of attachment of infectious virus through the interaction of the hemagglutinin molecule and live mammalian and avian cells and the subsequent infection of these cells. To this end, influenza viruses of various HA subtypes were constructed that either carry the green fluorescent protein (GFP) instead of the neuraminidase protein, or that express GFP in the cytoplasm of infected cells. The HA genes were derived from influenza viruses A/PR/8/34 (H1N1), A/Netherlands/178/95 (H3N2) and A/Vietnam/1194/04 (H5N1). Using these pairs of viruses, attachment and post-attachment events in the virus replication cycle can be distinguished. In general, the expression of NeuAc(alpha 2-3)Gal or NeuAc(alpha 2-6)Gal receptors on the cells tested corresponded with the attachment of the viruses that were studied with respect to predicted receptor specificity. Virus attachment was not always predictive for efficient infection of the cells. (c) 2007 Elsevier B.V. All rights reserved.