Single-Cell Virology: On-Chip, Quantitative Characterization of the Dynamics of Virus Spread from One Single Cell to Another

• Published in: Viruses Vol. 16; no. 11; p. 1659
• Main Authors: Liu, Wu, Wilke, Claus O., Arnold, Jamie J., Sotoudegan, Mohamad S., Cameron, Craig E.
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 01.11.2024; MDPI
• Subjects: Analysis; Autophagy; Cell culture; Cell Line; Distribution; Enteroviruses; Host-virus relationships; Humans; Infections; Kinases; Lab-On-A-Chip Devices; Microfluidics; non-lytic spread; Poliovirus; Poliovirus - physiology; Proteins; secretory autophagy; Single-Cell Analysis - methods; single-cell virology; Uncoating; Unicellular organisms; viral infection dynamics; Viral infections; viral spread dynamics; Virology; Virology - methods; Virus Internalization; Virus Replication; Viruses; United States; viral spread dynamics; enteroviruses; non-lytic spread; viral infection dynamics; single-cell virology; secretory autophagy
• ISSN: 1999-4915; 1999-4915
• DOI: 10.3390/v16111659

Virus spread at the single-cell level is largely uncharacterized. We have designed and constructed a microfluidic device in which each nanowell contains a single, infected cell (donor) and a single, uninfected cell (recipient). Using a GFP-expressing poliovirus as our model, we observed both lytic and non-lytic spread. Donor cells supporting lytic spread established infection earlier than those supporting non-lytic spread. However, non-lytic spread established infections in recipient cells substantially faster than lytic spread and yielded higher rates of genome replication. While lytic spread was sensitive to the presence of capsid entry/uncoating inhibitors, non-lytic spread was not. Consistent with emerging models for non-lytic spread of enteroviruses using autophagy, reduction in LC3 levels in cells impaired non-lytic spread and elevated the fraction of virus in donor cells spreading lytically. The ability to distinguish lytic and non-lytic spread unambiguously will enable discovery of viral and host factors and host pathways used for non-lytic spread of enteroviruses and other viruses as well.