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Viruses 2018, 10(5), 220; https://doi.org/10.3390/v10050220

A New Approach to Assessing HSV-1 Recombination during Intercellular Spread

Department of Microbiology & Immunology, Montana State University, Bozeman, MT 59717, USA
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Received: 9 March 2018 / Revised: 17 April 2018 / Accepted: 20 April 2018 / Published: 25 April 2018
(This article belongs to the Special Issue Viral Recombination: Ecology, Evolution and Pathogenesis)
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Abstract

The neuroinvasive Herpes simplex virus type 1 (HSV-1) utilizes intergenomic recombination in order to diversify viral populations. Research efforts to assess HSV-1 recombination are often complicated by the use of attenuating mutations, which differentiate viral progeny but unduly influence the replication and spread. In this work, we generated viruses with markers that allowed for classification of viral progeny with limited attenuation of viral replication. We isolated viruses, harboring either a cyan (C) or yellow (Y) fluorescent protein (FP) expression cassette inserted in two different locations within the viral genome, in order to visually quantify the recombinant progeny based on plaque fluorescence. We found that the FP marked genomes had a limited negative affect on the viral replication and production of progeny virions. A co-infection of the two viruses resulted in recombinant progeny that was dependent on the multiplicity of infection and independent of the time post infection, at a rate that was similar to previous reports. The sequential passage of mixed viral populations revealed a limited change in the distribution of the parental and recombinant progeny. Interestingly, the neuroinvasive spread within neuronal cultures and an in vivo mouse model, revealed large, random shifts in the parental and recombinant distributions in viral populations. In conclusion, our approach highlights the utility of FP expressing viruses in order to provide new insights into mechanisms of HSV-1 recombination. View Full-Text
Keywords: alphaherpesvirus; HSV-1; recombination; fluorescent protein; cell–cell spread; neuroinvasion; neuron culture; intravitreal injection alphaherpesvirus; HSV-1; recombination; fluorescent protein; cell–cell spread; neuroinvasion; neuron culture; intravitreal injection
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

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Law, G.A.; Herr, A.E.; Cwick, J.P.; Taylor, M.P. A New Approach to Assessing HSV-1 Recombination during Intercellular Spread. Viruses 2018, 10, 220.

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