Transfer and expression of the lacZ gene in rat brain neurons mediated by herpes simplex virus mutants

Three mutants of herpes simplex virus type 1 (HSV-1) were used to deliver and express the Escherichia coli lacZ gene in cells of the rat central nervous system. Because the lacZ gene was inserted in place of the genes encoding one of the immediate-early viral proteins ICP0 or ICP4 or the early viral protein thymidine kinase, these mutants were compromised or defective in their ability to replicate. All mutant vectors exhibited reduced pathogenesis in animals as compared to the wild type HSV-1 strain KOS. In all cases lacZ was under the control of immediate-early or early viral promoters that are active in the early phase of infection. Expression of beta-galactosidase was observed in cortical neurons following stereotactic inoculation of mutant viruses into adult rat brains; distinct patterns of expression were observed with each mutant vector. Injection of the ICP0 mutant in the frontal cortex and caudate nucleus resulted in beta-galactosidase expression in a substantial number of cells around the inoculation site and at some distance from it for 14 days, with maximum expression after 3 days. The ICP0 vector appeared to have reached the ipsilateral and contralateral cingulate cortex by retrograde transport. Following inoculations of the ICP4 and thymidine kinase vectors into the same brain regions, only a few cells in areas immediately adjacent to the injection track expressed beta-galactosidase and they did so for only a few days. These herpes virus-derived vectors provide a means for the in situ delivery and expression of specific genes in neurons in the central nervous system with little adverse effect on animals.