Spatiotemporal analysis of host cell modification during herpes simplex virus 1 replication

Abstract

Herpesviruses are large and complex viruses, and have a long evolutionary history with their host species. One poorly understood, but important, area in virus-host interaction is the extensive remodelling of intracellular organelles and morphology that occurs during active viral replication. We have constructed a recombinant reporter virus that allowed us to classify four distinct stages in the infection cycle of herpes simplex virus 1. Single-cell analysis of the impact of the viral load on replication dynamics demonstrates the utility of this timestamping method for tracking the infection cycle. High-resolution microscopy analysis of cellular structures in live and fixed cells in concert with our dual-fluorescent reporter virus have enabled us to generate a detailed overview over the spatial and temporal organisation of the cytoskeleton and organelles during viral replication. This provides the first systems-level analysis of the morphological changes involved in viral replication.Scherer et al.2 / 24

Single-cell Herpes Simplex Virus type-1 infection of neurons using drop-based microfluidics reveals heterogeneous replication kinetics

“…The dose dependence on viral gene expression and productive replication aligns with other work that observed dosedependent HSV-1 replication in human foreskin fibroblasts (40). Similarly, the effects of lower inoculating dose leading to slower kinetics of expression and low rates of productive replication have been observed during HSV-1 infection of other non-neuronal cells (33,40). Notably, we were able to observe single-cell infection of neurons for longer observation periods.…”

“…Importantly, all RFP positive neurons were detectably YFP positive before 6 hpi, suggesting that early viral transcription is more likely to elicit productive viral replication. The dose dependence on viral gene expression and productive replication aligns with other work that observed dosedependent HSV-1 replication in human foreskin fibroblasts (40). Similarly, the effects of lower inoculating dose leading to slower kinetics of expression and low rates of productive replication have been observed during HSV-1 infection of other non-neuronal cells (33,40).…”

Combining sample expansion and light sheet microscopy for the volumetric imaging of virus-infected cells with super-resolution

“…Expansion microscopy is a technique that has not yet been widely explored in virus research [ 40 ]. However, we show that it provides unprecedented detail on the interaction and localisation of virus particles with subcellular organelles.…”