Partial rescue of herpes simplex virus neurovirulence with a 3.2 kb cloned DNA fragment.

A herpes simplex virus (HSV) intertypic recombinant (RE6) has been previously shown to be non-neurovirulent following direct intracranial inoculation of mice. An in vitro recombination/in vivo selection strategy was employed to further characterize the gene or genes responsible for the avirulence of this mutant. It was found that RE6 could be converted to a lethal virus by incorporation of wild-type HSV-1 sequences mapping between 0.698 and 0.721 map units. Restriction endonuclease and Southern blot analysis revealed that viral recombinants which incorporated at least part of the cloned 17syn+ sequences were selected by passage in mouse brains in vivo. The recombinants generated with this fragment were at least 50-fold more neuroviurlent than RE6, as determined by the plaque forming unit to lethal dose 50% assay. Further, they displayed a significant increase in ability to replicate in mouse brain tissue following intracranial inoculation. However, these recombinants did not display a replication advantage over RE6 in cultured mouse cells at 38.5 degrees C. Thus, the defect present within this region of the RE6 genome specifically affects replication in the nervous system. In the accompanying paper we analyze the RNA transcription and DNA sequence in this portion of the RE6 and parental strain genomes.