Evaluation of conditions and mechanisms of alphaherpesvirus superinfection exclusion

Abstract

Superinfection exclusion (SIE) is a natural phenomenon where one virus prevents subsequent entry of another virus. Studies have shown its impact on viral replication and reproduction of many different viruses, where exclusion targets specific parts of the virus' lifecycle. Alphaherpesviruses, specifically pseudorabies virus (PRV) and Herpes Simplex virus type 1 (HSV-1), have demonstrated at least two forms of SIE. However, the form of SIE that occurs early in viral replication remains of interest due to both its timing and unknown mechanisms of regulation. Research on this topic will stimulate development of vaccines in global health by providing targets for disruption of alphaherpesvirus infection and information on unknown aspects of alphaherpesvirus infection. We have developed two models for assessing early SIE for alphaherpesviruses: a fluorescent reporter model to quantify virions within the cell. Both models contributed to data that determined our early SIE is influenced by multiplicity of infection (MOI) for both the primary and secondary virions. Imaging flow cytometry was utilized in conjunction with fluorescent microscopy as a possible means to study early SIE in large population samples. Subsequent data from my experiments indicates that cellular factors like cellular receptors, clathrin, and actin arrangement had minimal influence on early SIE of alphaherpesviruses. However, the results from capsid trafficking model in combination with experiments involving heparin and bortezomib indicate that early SIE differs in PRV and HSV-1. Data indicates that PRV SIE inhibits the step of viral entry/fusion, while HSV-1 SIE disrupts in a post-entry manner. Data obtained from this dissertation indicates that early SIE influences alphaherpesviruses differently and presents possible means to study antiviral techniques and methods.