Enhanced expression of an α2,6-linked sialic acid on MDCK cells improves isolation of human influenza viruses and evaluation of their sensitivity to a neuraminidase inhibitor
Enhanced expression of an α2,6-linked sialic acid on MDCK cells improves isolation of human influenza viruses and evaluation of their sensitivity to a neuraminidase inhibitor
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DOI：
10.1128/jcm.43.8.4139-4146.2005
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发表时间：
2005-08-01
影响因子：
9.4
通讯作者：
Kawaoka, Y
中科院分区：
文献类型：
--
作者：
Hatakeyama, S;Sakai-Tagawa, Y;Kawaoka, Y
神经氨酸酶（NA）抑制剂治疗流感病毒感染的广泛使用要求密切监测耐药变异。培养细胞不能提供一种可靠的方法来评价人流感病毒分离株对NA抑制剂的敏感性。也就是说，这种病毒在细胞系中的生长（例如，Madin-Darby犬肾[MDCK]细胞）不受这些药物的抑制，即使它们的唾液酸酶活性对药物敏感。Matrosovich等人（J. Virol. 77：8418-8425，2003）显示，基于对有限数量的病毒的研究，过表达人β-半乳糖苷α 2，6-唾液酸转移酶I（ST 6 Ga 1 I）基因的MDCK细胞系具有评估人流感病毒分离株对NA抑制剂的敏感性的潜力。在此，我们询问流感病毒的临床分离株是否对表达ST 6 Gal I基因的MDCK细胞系中的NA抑制剂（奥司他韦）普遍敏感。病毒对奥司他韦的敏感性与病毒唾液酸酶对化合物的敏感性相关，证明了这种修饰的细胞系用于检测NA受体耐药病毒的潜在效用。此外，在ST 6 Gal I过表达的细胞中，人流感病毒的生长比MDCK细胞中高2个对数。我们得出结论，人ST 6 Ga 1 I表达MDCK细胞系不仅可用于评估它们对NA抑制剂的敏感性，而且可用于从临床样品中分离流感病毒。
The extensive use of neuraminidase (NA) inhibitors to treat influenza virus infections mandates close monitoring for resistant variants. Cultured cells do not provide a reliable means of evaluating the susceptibility of human influenza virus isolates to NA inhibitors. That is, the growth of such viruses in cell lines (e.g., Madin-Darby canine kidney [MDCK] cells) is not inhibited by these drugs, even though their sialidase activity is drug-sensitive. Matrosovich et al. (J. Virol. 77:8418-8425, 2003) showed that an MDCK cell line overexpressing the human beta-galactoside alpha 2,6-sialyltransferase I (ST6Ga1 I) gene has the potential to assess the sensitivity of human influenza virus isolates to NA inhibitors, based on studies with a limited number of viruses. Here, we asked whether clinical isolates of influenza virus are universally sensitive to an NA inhibitor (oseltamivir) in an MDCK cell line expressing the ST6Gal I gene. The sensitivity of viruses to oseltamivir correlated with the sensitivity of viral sialidase to the compound, demonstrating the potential utility of this modified cell line for detecting NA inhibitor-resistant viruses. Moreover, in ST6Gal I-overexpressing cells, the growth of human influenza viruses was up to 2 logs higher than in MDCK cells. We conclude that the human ST6Ga1 I-expressing MDCK cell line is useful not only for evaluating their sensitivity to NA inhibitors, but also for isolation of influenza viruses from clinical samples.