The characteristics and antigenic properties of recently emerged subclade 3C.3a and 3C.2a human influenza A(H3N2) viruses passaged in MDCK cells.
The characteristics and antigenic properties of recently emerged subclade 3C.3a and 3C.2a human influenza A(H3N2) viruses passaged in MDCK cells.
复制标题
DOI：
10.1111/irv.12447
复制
发表时间：
2017-05
影响因子：
4.4
通讯作者：
McCauley JW
中科院分区：
文献类型：
--
作者：
Lin Y;Wharton SA;Whittaker L;Dai M;Ermetal B;Lo J;Pontoriero A;Baumeister E;Daniels RS;McCauley JW
在2014-2015年北半球流感季节，两种新的甲型H3N2型流感病毒变得突出。这些病毒的HA糖蛋白显示出以前与受体结合特性变化相关的序列变化。为了解决这些变化如何影响病毒传播，在传统的MDCK细胞和MDCK-SIAT1细胞中分离和繁殖病毒，并在每一代中对病毒的人类受体表达增强的细胞进行分析。病毒在常规MDCK细胞和MDCK-SIAT1细胞中传代时进行基因序列分析。通过使用豚鼠、火鸡和人类的红细胞进行血凝试验，检测了与病毒传播相关的受体识别的变化。微中和试验用于确定传代获得性氨基酸替代和多态如何影响病毒抗原性。病毒感染MDCK-SIAT1细胞的效率高于常规MDCK细胞。与MDCK-SIAT1细胞相比，3C.2a和3C.3a亚类病毒在传代过程中显示出更大的序列变化，在HA和NA糖蛋白中都可以看到氨基酸替换。然而，在MDCK-SIAT1细胞中，低稀释度的病毒传代显示继续传代的传染性降低。目前的H3N2病毒应该在MDCK-SIAT1细胞系中培养，以保持病毒的忠实复制，并在适当的感染多样性下保持传染性。
Two new subclades of influenza A(H3N2) viruses became prominent during the 2014‐2015 Northern Hemisphere influenza season. The HA glycoproteins of these viruses showed sequence changes previously associated with alterations in receptor‐binding properties. To address how these changes influence virus propagation, viruses were isolated and propagated in conventional MDCK cells and MDCK‐SIAT1 cells, cells with enhanced expression of the human receptor for the virus, and analysed at each passage. Gene sequence analysis was undertaken as virus was passaged in conventional MDCK cells and MDCK‐SIAT1 cells. Alterations in receptor recognition associated with passage of virus were examined by haemagglutination assays using red blood cells from guinea pigs, turkeys and humans. Microneutralisation assays were performed to determine how passage‐acquired amino acid substitutions and polymorphisms affected virus antigenicity. Viruses were able to infect MDCK‐SIAT1 cells more efficiently than conventional MDCK cells. Viruses of both the 3C.2a and 3C.3a subclades showed greater sequence change on passage in conventional MDCK cells than in MDCK‐SIAT1 cells, with amino acid substitutions being seen in both HA and NA glycoproteins. However, virus passage in MDCK‐SIAT1 cells at low inoculum dilutions showed reducing infectivity on continued passage. Current H3N2 viruses should be cultured in the MDCK‐SIAT1 cell line to maintain faithful replication of the virus, and at an appropriate multiplicity of infection to retain infectivity.