Herpes simplex virus latency and reactivation in isolated rat sensory neurons

Abstract

An in vitro herpes simplex virus type 1 (HSV-1) latency model has been established using neurons isolated from dissociated rat fetus sensory ganglia as the host cell. Rat fetal neuron cells were pretreated for 24 hr at 37 degrees with (E)-5-(2-bromovinyl)-2'-deoxyuridine and human leukocyte interferon, infected with HSV-1 (approximately 2.5 plaque-forming units/cell), and treated for 7 days with the same inhibitor combination. Infectious HSV-1 became undetectable 3 days postinfection and remained undetectable during the remainder of the inhibitor treatment. After removal of inhibitors on day 7, infectious virus remained undetectable for 2-7 days; subsequently, virus replication ensued and neuronal cells were destroyed. Incubation of inhibitor-treated, infected neuron cells at 40.5 degrees after removal of inhibitors resulted in extension of the latent period to at least 15 days. HSV-1 was reactivated from latently infected neurons by reducing the incubation temperature from 40.5 to 37 degrees and virus-specific cytopathology was observed in neurons within 96 hr after reducing temperature. This in vitro model system will provide the first system to analyze, in a primary cell type of neuronal origin, the state of the HSV genome during establishment and maintenance of the latent state and during virus reactivation.