Replication of the herpes simplex virus type 1 RL1 mutant 1716 in primary neuronal cell cultures--possible relevance to use as a viral vector.
The herpes simplex virus type 1 (HSV-1) RL1 deletion mutant 1716 has properties that make it a promising candidate as a viral vector for gene therapy in the human nervous system. These properties include its ability to spread along neural pathways and establish a latent infection in post-mitotic neurons, while retaining a non-virulent phenotype in vivo and an inability to cause a lytic infection in stationary or fully differentiated cells. In this study, we used viral replication assays and indirect immunofluorescence to investigate the ability of 1716 to bind to, enter, express genes and produce progeny virus in dissociated neuronal cell cultures prepared from rat hippocampal, medial septal and dorsal root ganglion (DRG) tissues and in primary rat astrocyte cultures. Both heterogeneous cultures and those that had been enriched for neurons were employed. Following both low and high multiplicities of virus infection, the behaviour of 1716 was compared with its wild-type parent HSV-1 strain 17 in these cultures. It was found that the growth of 1716 was significantly impaired compared to wild type HSV-1, with these differences being magnified at lower multiplicities of viral infection as well as in neuron-enriched cultures: this impairment is likely to be due to decreased replication, as immunofluorescence assays showed that 1716 bound to, entered and expressed genes in all neuronal cell types and astrocytes with similar efficiency to the wild type virus. This ability of 1716 to enter and express genes in different neuronal populations demonstrates its potential suitability as a viral vector.
Kennedy
P G
PG
Department of Neurology, South Glasgow University Hospitals NHS Trust, Institute of Neurological Sciences, Govan Road, G51 4TF, Glasgow, UK. pgkle@clinmed.gla.ac.uk
Gairns
J
J
MacLean
A R
AR
D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000378:metabolism / Q000821:virology; D018929:Cell Culture Techniques / Q000379:methods / Q000639:trends; D002478:Cells, Cultured / Q000166:cytology / Q000378:metabolism / Q000821:virology*; D005333:Fetus; D005455:Fluorescent Antibody Technique; D015316:Genetic Therapy / Q000379:methods* / Q000639:trends*; D005822:Genetic Vectors / Q000378:metabolism*; D018259:Herpesvirus 1, Human / Q000235:genetics*; D009154:Mutation / Q000235:genetics; D009474:Neurons / Q000166:cytology / Q000378:metabolism / Q000821:virology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D014779:Virus Replication / Q000235:genetics*