Consensus and variations in cell line specificity among human metapneumovirus strains
Consensus and variations in cell line specificity among human metapneumovirus strains
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DOI：
10.1371/journal.pone.0215822
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发表时间：
2019-04-23
期刊：
影响因子：
3.7
通讯作者：
Takeda, Makoto
中科院分区：
文献类型：
--
作者：
Nao, Naganori;Sato, Ko;Takeda, Makoto
人偏肺病毒（Human metapneumovirus，HMPV）是引起人类急性呼吸道感染的重要病原体，但由于HMPV只能在有限的细胞系中复制，致细胞病变（cytopathic effect，CPE）轻微，直到2001年才被发现。为了促进对HMPV的研究，一些研究小组已经产生了表达绿色荧光蛋白（GFP）的重组HMPV株（HMPVGFP）。然而，越来越多的证据使对HMPV细胞系特异性的理解变得复杂，因为它似乎在HMPV毒株之间存在显著差异。此外，最近已经检测到在G基因中具有180个核苷酸重复的独特A2 b进化枝HMPV毒株（HMPV A2 b（180 nt-dup）毒株）。在这项研究中，我们重新评估和比较了HMPV临床分离株的细胞系特异性，包括新的HMPV A2 b（180 nt-dup）株，和6个重组HMPV GFP株，包括新产生的重组HMPV A2 b（180 nt-dup）株，MG 0256-EGFP。我们的数据表明，VeroE 6和LLC-MK2细胞通常显示出对任何临床分离株和重组HMPVGFP菌株的最高感染性。其他人源细胞系（BEAS-2B、A549、HEK 293、MNT-1和HeLa细胞）显示出一定水平的HMPV感染性，但这些显著低于VeroE 6和LLC-MK2细胞的感染性。此外，这些次优细胞系的感染性在HMPV毒株之间差异很大。这些变异与HMPV基因型、用于分离和繁殖的细胞系、特定基因组突变或G基因中的核苷酸重复无直接关系。因此，次优细胞系中的这些变异可能是特定HMPV毒株固有的。
Human metapneumovirus (HMPV) has been a notable etiological agent of acute respiratory infection in humans, but it was not discovered until 2001, because HMPV replicates only in a limited number of cell lines and the cytopathic effect (CPE) is often mild. To promote the study of HMPV, several groups have generated green fluorescent protein (GFP)-expressing recombinant HMPV strains (HMPVGFP). However, the growing evidence has complicated the understanding of cell line specificity of HMPV, because it seems to vary notably among HMPV strains. In addition, unique A2b clade HMPV strains with a 180-nucleotide duplication in the G gene (HMPV A2b(180nt-dup) strains) have recently been detected. In this study, we re-evaluated and compared the cell line specificity of clinical isolates of HMPV strains, including the novel HMPV A2b(180nt-dup) strains, and six recombinant HMPVGFP strains, including the newly generated recombinant HMPV A2b(180nt-dup) strain, MG0256-EGFP. Our data demonstrate that VeroE6 and LLC-MK2 cells generally showed the highest infectivity with any clinical isolates and recombinant HMPVGFP strains. Other human-derived cell lines (BEAS-2B, A549, HEK293, MNT-1, and HeLa cells) showed certain levels of infectivity with HMPV, but these were significantly lower than those of VeroE6 and LLC-MK2 cells. Also, the infectivity in these suboptimal cell lines varied greatly among HMPV strains. The variations were not directly related to HMPV genotypes, cell lines used for isolation and propagation, specific genome mutations, or nucleotide duplications in the G gene. Thus, these variations in suboptimal cell lines are likely intrinsic to particular HMPV strains.