Comparison of SARS-CoV-2 Evolution in Paediatric Primary Airway Epithelial Cell Cultures Compared with Vero-Derived Cell Lines.
Comparison of SARS-CoV-2 Evolution in Paediatric Primary Airway Epithelial Cell Cultures Compared with Vero-Derived Cell Lines.
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DOI：
10.3390/v14020325
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发表时间：
2022-02-05
期刊：
影响因子：
--
通讯作者：
On Behalf Of The Breathing Together Investigators
中科院分区：
文献类型：
--
作者：
Bamford CGG;Broadbent L;Aranday-Cortes E;McCabe M;McKenna J;Courtney DG;Touzelet O;Ali A;Roberts G;Lopez Campos G;Simpson D;McCaughey C;Fairley D;Mills K;Power UF;On Behalf Of The Breathing Together Investigators
SARS-CoV-2可以有效地感染儿童和成人，尽管发病率和死亡率与宿主年龄的增加和合并症的存在呈正相关。SARS-CoV-2继续适应人群，产生了几种具有新特性的关注变体（VOC），如Alpha和Delta。然而，驱动SARS-CoV-2适应性和儿科队列演变的因素仍然很少探索。在这里，我们提供的证据表明，病毒和宿主因素的合作，以塑造SARS-CoV-2基因型和表型的变化，在原代气道细胞培养来自儿童。通过病毒全基因组测序，我们探索了两种SARS-CoV-2的VOC前临床分离株在儿科分化良好的原代鼻上皮细胞（WD-PNEC）培养物中传代过程中以及在未修饰的Vero细胞系中随时间推移的遗传多样性变化。我们确定了一个一致的，丰富的遗传多样性在体外产生的，其中的变体可以迅速上升到近固定在两个通道。在分离株中，SARS-CoV-2的进化依赖于宿主细胞，与Vero（E6）细胞相比，儿科WD-PNEC显示出减少的多样性。然而，突变在菌株之间不共享。此外，在WD-PNEC上比较两种Vero生长的分离株，发现在Spike中多碱基切割位点（PBCS）的丢失导致了显著的生长衰减，而在Nsp 12（T293 I）、Spike（P812 R）和Orf 7a截短突变的菌株在WD-PNEC中仍然存活。总之，我们的工作表明，前VOC SARS-CoV-2有效地感染儿科呼吸道上皮细胞，其进化受到抑制相比，Vero（E6）细胞，类似于成人细胞的情况。我们强调了SARS-CoV-2的显著遗传可塑性，同时揭示了病毒和宿主细胞因子在塑造病毒进化和最终在人类呼吸道上皮中的适应性方面的合作的影响作用。
SARS-CoV-2 can efficiently infect both children and adults, albeit with morbidity and mortality positively associated with increasing host age and presence of co-morbidities. SARS-CoV-2 continues to adapt to the human population, resulting in several variants of concern (VOC) with novel properties, such as Alpha and Delta. However, factors driving SARS-CoV-2 fitness and evolution in paediatric cohorts remain poorly explored. Here, we provide evidence that both viral and host factors co-operate to shape SARS-CoV-2 genotypic and phenotypic change in primary airway cell cultures derived from children. Through viral whole-genome sequencing, we explored changes in genetic diversity over time of two pre-VOC clinical isolates of SARS-CoV-2 during passage in paediatric well-differentiated primary nasal epithelial cell (WD-PNEC) cultures and in parallel, in unmodified Vero-derived cell lines. We identified a consistent, rich genetic diversity arising in vitro, variants of which could rapidly rise to near fixation within two passages. Within isolates, SARS-CoV-2 evolution was dependent on host cells, with paediatric WD-PNECs showing a reduced diversity compared to Vero (E6) cells. However, mutations were not shared between strains. Furthermore, comparison of both Vero-grown isolates on WD-PNECs disclosed marked growth attenuation mapping to the loss of the polybasic cleavage site (PBCS) in Spike, while the strain with mutations in Nsp12 (T293I), Spike (P812R) and a truncation of Orf7a remained viable in WD-PNECs. Altogether, our work demonstrates that pre-VOC SARS-CoV-2 efficiently infects paediatric respiratory epithelial cells, and its evolution is restrained compared to Vero (E6) cells, similar to the case of adult cells. We highlight the significant genetic plasticity of SARS-CoV-2 while uncovering an influential role for collaboration between viral and host cell factors in shaping viral evolution and ultimately fitness in human respiratory epithelium.
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影响因子：
64.5
作者：
Hou, Yixuan J.;Okuda, Kenichi;Baric, Ralph S.
通讯作者：
Baric, Ralph S.
DOI：
10.1056/nejmoa2022926
发表时间：
2020-11-19
期刊：
The New England journal of medicine
影响因子：
--
作者：
RECOVERY Collaborative Group;Horby P;Mafham M;Linsell L;Bell JL;Staplin N;Emberson JR;Wiselka M;Ustianowski A;Elmahi E;Prudon B;Whitehouse T;Felton T;Williams J;Faccenda J;Underwood J;Baillie JK;Chappell LC;Faust SN;Jaki T;Jeffery K;Lim WS;Montgomery A;Rowan K;Tarning J;Watson JA;White NJ;Juszczak E;Haynes R;Landray MJ
通讯作者：
Landray MJ
DOI：
10.1164/rccm.201304-0750oc
发表时间：
2013-10-01
影响因子：
24.7
作者：
Guo-Parke, Hong;Canning, Paul;Power, Ultan F.
通讯作者：
Power, Ultan F.
DOI：
10.1099/jgv.0.001453
发表时间：
2020-09
期刊：
The Journal of general virology
影响因子：
--
作者：
Ogando NS;Dalebout TJ;Zevenhoven-Dobbe JC;Limpens RWAL;van der Meer Y;Caly L;Druce J;de Vries JJC;Kikkert M;Bárcena M;Sidorov I;Snijder EJ
通讯作者：
Snijder EJ
影响因子：
46.9
作者：
Loske, J.;Roehmel, J.;Lehmann, I
通讯作者：
Lehmann, I