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Herpes Simplex Virus-2 Reactivation from Viral Latency in Autonomic Ganglia

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[Technical Report, Doctoral Thesis]

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Herpes Simplex Virus-2 is a worldwide, major human pathogen that causes recurrent disease over the lifetime of its host. According to current dogma, HSV-2reactivation occurs from the sensory, sacral dorsal root ganglia. However, a growing body of evidence reports the virus latent in another branch of the peripheral nervous system, the autonomic ganglia. It is unclear whether spontaneous reactivation is occurring in these additional sites, potentially contributing to recurrent genital skin lesions and observed clinical symptoms like urinary retention or responsible for subclinical reactivations in the dermis. Due to the small size of virions and the rare spontaneous event of reactivation, a fluorescent herpes simplex virus is an invaluable tool for localizing virus in cells. Unfortunately, available fluorescent mutants lack the required sufficient genetic stability viior normal kinetics of replication and reactivation. We hypothesized that by using a brighter, non-dimerizing fluorescent protein, we could engineer a functional mutant virus to begin to address some of the key questions related to reactivation of HSV-2. We constructed a fluorescent HSV-2 strain by fusing the fluorescent mNeonGreen protein to the VP26 minor capsid protein. This mutant virus displayed normal replication and in vivo recurrence phenotypes, providing an improved tool for investigating reactivation from latency. In order to explore the full extent of the tropism of reactivation we used the guinea pig vaginal infection model that develops spontaneous recurrent genital skin lesions and signs of autonomic dysfunction in the form of urinary retention and constipation.

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  • Microbiology

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[A, Approved For Public Release]