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, Jojoba 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 Catheters and Sheaths - Intermittent enteroviruses and other viruses as well.