BARCODED HERPES SIMPLEX VIRUS-1 RECOMBINANTS REVEAL VARIABILITY OF INFECTION IN SINGLE CELLS

Single cell variation during herpesviruses infection may contribute to different outcomes of infection. Here, we developed a method to assess the variation in the number of incoming viral genomes replicating (NOIVGR) in single cells, using fourteen barcoded, fluorescence-expressing, viral recombinants. This method provides the NOIVGR of the individual cell tested. Our results indicate the average NOIVGR per cell, as well as its variability among cells. In individual cells, this number correlates with the expressed fluorescence from these viruses. The relative fluorescence was found to correlate with viral gene expression. A multiple linear regression model was developed to predict the NOIVGR in a single cell according to the multiplicity of infection, fluorescence intensity and side-scattered levels. Our results link parameters of individual cells with the NOIVGR. We propose that the NOIVGR is dependent on cellular determinants and feeds back to the outcome of infection in that cell.