Long-term expression in sensory neurons in tissue culture from herpes simplex virus type 1 (HSV-1) promoters in an HSV-1-derived vector.
Amplicons, defective herpes simplex virus type 1 (HSV-1) vectors, were constructed to use four HSV-1 promoters, from the immediate-early (IE) 1 IE 3, IE 4/5, and late glycoprotein C (gC) genes, to regulate expression of the Escherichia coli lacZ gene, encoding beta-galactosidase, and packaged into infectious particles. Infection of sensory neurons in vitro with amplicons containing the IE 1, IE 3, or IE 4/5 promoter resulted in stable long-term expression of beta-galactosidase from 2 to 10 weeks after gene transfer. The number of neurons expressing beta-galactosidase was not changed by treatments previously shown to produce reactivation of latent HSV-1. In addition, the latency-associated transcript was detected in many of the same neurons that expressed beta-galactosidase, indicating that the viral IE promoters in the amplicons can function in the same neurons that harbor latent virus. Delivery of beta-galactosidase protein directly into neurons by microinjection indicated that the half-life for histochemical detection of beta-galactosidase was between 24 and 48 h, suggesting that the persistence of beta-galactosidase histochemical staining cannot be explained by the stability of the reporter protein alone. In contrast to the IE promoters, the gC promoter of the late gene class did not support long-term expression of beta-galactosidase; instead, beta-galactosidase was detected in only a few neurons per culture at 2 weeks after infection, and superinfection with wild-type HSV-1 did not increase the level of expression from the gC promoter. These results suggest that the HSV-1 IE promoters in the amplicons are not subject to the promoter inactivation that occurs with many types of virus vectors and that the IE promoters in the context of the amplicon avoid the promoter inactivation observed from the same promoters in the HSV-1 genome during latency.
Smith
R L
RL
Department of Neurology and Pediatrics, University of Colorado Health Sciences Center, Denver, USA.
Geller
A I
AI
Escudero
K W
KW
Wilcox
C L
CL
D000818:Animals; D046508:Culture Techniques; D004279:DNA, Viral; D005727:Ganglia, Spinal / Q000166:cytology / Q000821:virology; D005784:Gene Amplification; D017781:Genes, Immediate-Early; D005822:Genetic Vectors; D018259:Herpesvirus 1, Human / Q000235:genetics* / Q000502:physiology; D009475:Neurons, Afferent / Q000821:virology*; D011401:Promoter Regions, Genetic; D051381:Rats; D017735:Virus Latency; D001616:beta-Galactosidase / Q000235:genetics
D004279:DNA, Viral; D001616:beta-Galactosidase