Establishment of Latency in Human Immortalized Dorsal Root Ganglia with the Neurotropic 1lsyn+ Strain of Herpes Simplex Virus Type 1
Blanchett, Reid E.
MetadataShow full item record
Herpes simplex virus type 1 is present in approximately 80% of the American population and is the causative agent of herpetic keratitis, viral encephalitis, and labial lesions. An FISV-1 infection is characterized by its three-stage lifecycle: the initial lytic infection, the establishment of latency, and the reactivation of the virus. Though lytic infections are quite well understood, much less is known about HSV-1 latency and reactivation. This disparity is partially due to a lack of adequate models to study the processes. The current in vivo and in vitro systems that exist are either inefficient, or inapplicable to human molecular physiology. This study establishes a latency-model in human immortalized dorsal root ganglia with the neurotropic HSV-1 17syn+ strain. The hallmarks of in vitro latency were demonstrated with viral plaque assays for infectious virus release, and quantitative real time PCR for viral genome detection in cells. X-gal staining of neurons latently infected with a 17syn+-based LAT (latency associated transcript) promoter-Bgal reporter virus validated that viral genomes present in cells were functional and capable of LAT expression. By demonstrating two of the three hallmarks of experimental latency, the presented system fills the current gap in latency and reactivation models: a latency system with the potential to reactivate that is both cost effective and relevant to human molecular physiology.