Hope Thomas, Becker Mark, Martin-Sancho Laura, Simons Lacy, McRaven Michael, Chanda Sumit, Hultquist Judd
Northwestern University.
Imperial College London.
Res Sq. 2023 Sep 8:rs.3.rs-3246773. doi: 10.21203/rs.3.rs-3246773/v1.
SARS-CoV-2 initiates infection in the conducting airways, which rely on mucocilliary clearance (MCC) to minimize pathogen penetration. However, it is unclear how MCC impacts SARS-CoV-2 spread after infection is established. To understand viral spread at this site, we performed live imaging of SARS-CoV-2 infected differentiated primary human bronchial epithelium cultures for up to 9 days. Fluorescent markers for cilia and mucus allowed longitudinal monitoring of MCC, ciliary motion, and infection. The number of infected cells peaked at 4 days post-infection in characteristic foci that followed mucus movement. Inhibition of MCC using physical and genetic perturbations limited foci. Later in infection, MCC was diminished despite relatively subtle ciliary function defects. Resumption of MCC and infection spread after mucus removal suggests that mucus secretion mediates this effect. We show that MCC facilitates SARS-CoV-2 spread early in infection while later decreases in MCC inhibit spread, suggesting a complex interplay between SARS-CoV-2 and MCC.
严重急性呼吸综合征冠状病毒2（SARS-CoV-2）在传导气道引发感染，传导气道依靠黏液纤毛清除（MCC）来尽量减少病原体的侵入。然而，尚不清楚在感染确立后MCC如何影响SARS-CoV-2的传播。为了解该部位的病毒传播情况，我们对感染SARS-CoV-2的分化原代人支气管上皮细胞培养物进行了长达9天的实时成像。用于纤毛和黏液的荧光标记物能够对MCC、纤毛运动和感染进行纵向监测。感染细胞数量在感染后第4天达到峰值，出现在跟随黏液移动的特征性病灶中。使用物理和基因干扰对MCC进行抑制会限制病灶。在感染后期，尽管纤毛功能存在相对细微的缺陷，但MCC仍有所减弱。去除黏液后MCC的恢复和感染传播表明黏液分泌介导了这种效应。我们发现，MCC在感染早期促进SARS-CoV-2传播，而后期MCC的减少则抑制传播，这表明SARS-CoV-2与MCC之间存在复杂的相互作用。
