INFECTION AND REPLICATION OF HERPES-SIMPLEX VIRUS TYPE-1 IN AN ORGANOTYPIC EPITHELIAL CULTURE SYSTEM

We have used the organotypic culture system as a model to study the in itial infectious process and spread of herpes simplex virus type 1 (HS V-1) in fully stratified and differentiated human epithelial tissue. T he growth kinetics of HSV-1 were determined in organotypic tissues of human epidermal or ectocervical origin. Concurrently, we followed the spread of HSV-1 by immunostaining thin sections of infected organotypi c tissue. After HSV-1 was applied to the top cornified epithelial laye r, virus penetrated to the basal layer of replicating epithelium and g rew to high titers. The virus was limited in its spread in that not al l cells within the tissue had demonstrable infection. A ribonucleotide reductase mutant, ICP6 Delta, could infect and replicate in basal lay ers of the organotypic tissues. However, we found that spread was limi ted in, and to, the basal cell layer. Peak ICP6 Delta titers were 100- fold less than in cultures infected with wild-type HSV-1. Studies of H SV mutants should allow us to further define the role of specific vira l genes which are associated with infection and spread in a tissue cul ture system that mimics the initial portal of entry for certain HSV in fections. (C) 1997 academic Press.