POSTER TITLE

Zoe Anderson

Abstract
Introduction
Herpes Simplex Virus (HSV) is widespread among the human population infecting

somewhere between 70-90% of people (Whitley). The virus is contracted on the body surface

where it infects and replicates in epithelial cells. From there, it spreads into sensory neurons.

This leads to lifelong infection due to its ability to establish latency in neurons of the host. Viral

DNA exists in the nucleus as a circular extra-chromosomal DNA. (Knipe et al. 2008). Latency

of the virus is defined as a lack of viral protein production as no protein is detected during this

time. The genes required for translation and transcription of lytic proteins are repressed in the

neuronal nucleus (Bloom et al. 2010). There is however production of micro-RNAs that have

been transcribed from latency-associated transcripts (LATS). Yet, HSV can periodically switch to

active viral replication in response to physiological signals (Fields Virology). The exact way in

which latency is maintained and how the virus enters the lytic cycle is still not fully understood.

It has been shown previously that binding of nerve growth factor (NGF) to the TrkA

receptor tyrosine kinase to activate neuronal phosphatidylinositol 3-kinase (PI3)/Akt signaling is

key for maintaining HSV latency (Camerena et al. 2010). Interruption of this cascade leads to

reactivation in neuronal cell cultures. More recent study has shown that local mTOR signaling, a

target of the PI3/Akt pathway, regulates latency. Quiescent cell cultures treated with mTOR

specific inhibitors were found to contain infectious virus. 4E-BP, a translational repressor, is one

of the main targets of mTOR signaling that controls the expression of viral genomes. 4E-BP is
inhibited by mTORC1 signaling. When this signaling is interrupted or 4E-BP is hyper-

phosphorylated, reactivation in cell cultures occurs. However, this is not true of p70 S6 kinase,

the other translational substrate of mTORC1 (Kobayashi et al. 2012).

The hybrid polar compound hexamethylene bisacetamide has often been used to increase

recovery of reactivated virus in explant and decrease the time for reactivation to occur.

(Bernstein et al 1987). The exact mechanism by which this happens is unknown but there are

various theories. In a study unrelated to HSV, HMBA was shown to inhibit activation of Akt

kinase in neurons (Dey et al.). This may prove to be the way HMBA is increasing recovered

virus during reactivation as we know this pathway is involved in controlling latency of the virus.

In this study, we show that PI3/Akt signaling is needed to control latency in explant and

HMBA disrupts this signaling. Unlike a quiescent neuronal cell culture, using an explant model

will allow us to see how host immunity affects the virus even in the presence of such inhibitors.

Swiss Webster mice were latently infected with 17syn+ HSV and the trigeminal ganglia were

removed and explanted. The explants were treated with a variety of inhibitors including HMBA

starting at the beginning of the PI3/Akt pathway becoming more specific. Reactivation was

induced and plaque assay .

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