Latency associated promoter transgene expression in the central nervous system after stereotaxic delivery of replication-defective HSV-1-based vectors.
The herpes simplex virus type 1 (HSV-1) latency associated promoter (LAP) has been shown to sustain long-term reporter gene expression within sensory neurones. Its activity within the CNS is, however, less well understood. In this study we characterise the activity of the LAP after stereotaxic delivery of recombinant HSV-1-based vectors to the brain. Two classes of vectors were utilised in these studies: (1) a replication-defective vector lacking the glycoprotein H and thymidine kinase genes, designated CS1, and (2) a virus mutant severely impaired for immediate-early (IE) gene expression which lacks functional VP16, ICP4 and ICP0 genes, designated in1388. Both vectors contain the LacZ gene under the control of the LAP. Following delivery of either vector to the striatum, beta-gal expression was detected within anatomically related CNS regions distal to the site of injection. At these sites the number of beta-gal-positive cells increased with time and remained stable up to 4 weeks p.i. beta-Gal expression could not be detected at the site of injection after delivery of CS1 but beta-gal expression within neurones located at this site was observed after delivery of in1388, indicating reduced toxicity of this severely disabled virus. Transgene expression decreased dramatically with both vectors at later time-points (>4 weeks after delivery), but PCR analysis demonstrated that viral genomes were stably maintained for up to 180 days following delivery, indicating that the loss of beta-gal-positive neurones was not likely to be due to a loss of vector-transduced cells. Moreover, after delivery of an equivalent virus to the rat striatum in situ hybridisation analysis showed a similar decrease in the number of neurones expressing the endogenous LATs with time. These data indicate that although the HSV-1 LAP can drive the expression of foreign genes in a variety of CNS neurones, in these cells there is a slow down-regulation of the viral promoter which eventually results in the loss of detectable transgene expression.
Scarpini
C G
CG
Division of Virology, Department of Pathology, University of Cambridge, Cambridge, UK.
May
J
J
Lachmann
R H
RH
Preston
C M
CM
Dunnett
S B
SB
Torres
E M
EM
Efstathiou
S
S
D000818:Animals; D001921:Brain / Q000201:enzymology*; D005260:Female; D015870:Gene Expression; D005822:Genetic Vectors / Q000008:administration & dosage*; D018259:Herpesvirus 1, Human / Q000235:genetics*; D007267:Injections; D007276:Injections, Intraventricular; D007763:Lac Operon; D009474:Neurons / Q000201:enzymology; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D051381:Rats; D017207:Rats, Sprague-Dawley; D013238:Stereotaxic Techniques; D013997:Time Factors; D019076:Transgenes; D017735:Virus Latency / Q000235:genetics*; D014779:Virus Replication; D001616:beta-Galactosidase / Q000032:analysis
D001616:beta-Galactosidase