Generation and characterization of recombinant influenza A (H1N1) viruses harboring amantadine resistance mutations
Generation and characterization of recombinant influenza A (H1N1) viruses harboring amantadine resistance mutations
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DOI：
10.1128/aac.49.2.556-559.2005
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发表时间：
2005-02-01
影响因子：
4.9
通讯作者：
Boivin, G
中科院分区：
文献类型：
--
作者：
Abed, Y;Goyette, N;Boivin, G
由于基质-（M）-2蛋白跨膜结构域内第26、27、30、31或34位的单氨基酸取代，已显示甲型流感病毒对金刚烷胺的耐药性的出现在治疗期间迅速发生。在这项研究中，反向遗传学被用来产生和表征重组甲型流感（H1N1）病毒携带L26 F，V27 A，A30 T，S31 N，G34 E，和V27 A/S31 N突变的M2基因。在空斑减少试验中，所有突变均赋予金刚烷胺抗性，药物浓度导致空斑数量减少50%（IC（50）），比野生型（WT）高154- 3，300倍。M2突变体在体外的噬斑大小和复制动力学实验的基础上，其复制能力没有损害。此外，在实验感染的小鼠中，所有突变体的毒性至少与WT相同，其中具有双重V27 A/S31 N突变的重组体获得的死亡率最高。这些发现可以帮助解释在临床环境中抗病毒压力期间，金刚烷胺耐药流感病毒的频繁出现和传播。
The emergence of resistance to amantadine in influenza A viruses has been shown to occur rapidly during treatment as a result of single-amino-acid substitutions at position 26, 27, 30, 31, or 34 within the transmembrane domain of the matrix-(M)-2 protein. In this study, reverse genetics was used to generate and characterize recombinant influenza A (H1N1) viruses harboring L26F, V27A, A30T, S31N, G34E, and V27A/S31N mutations in the M2 gene. In plaque reduction assays, all mutations conferred amantadine resistance, with drug concentrations resulting in reduction of plaque number by 50% (IC(50)s) 154- to 3,300-fold higher than those seen for the wild type (WT). M2 mutants had no impairment in their replicative capacities in vitro on the basis of plaque size and replication kinetics experiments. In addition, all mutants were at least as virulent as the WT in experimentally infected mice, with the highest mortality rate being obtained with the recombinant harboring a double V27A/S31N mutation. These findings could help explain the frequent emergence and transmission of amantadine-resistant influenza viruses during antiviral pressure in the clinical setting.