Changes in Corneal Innervation after HSV-1 Latency Established with Different Reactivation Phenotypes

Abstract

ABSTRACTPurpose of the study: We used a rabbit model infected with high phenotypic reactivators (HPRs) as well as recombinant HSV-1 (herpes simplex virus 1) with deletions to study their effect on corneal innervations after latency was established.Materials and methods: Corneas from noninfected New Zealand white rabbits were used to obtain the entire map of corneal innervation. Others were inoculated with the HSV-1 strains McKrae, 17Syn+, or recombinant mutants with glycoprotein K (gK) deletion, or with infected early protein 0 (ICP0) deletion. The animals were euthanized at 124 to 125 days postinfection and the corneas were immunostained with a mouse monoclonal anti-βIII tubulin antibody. Images were acquired with a fluorescence microscope and corneal sub-basal nerve density was calculated on the basis of the whole mount images. Differences between the HSV-infected eyes, and comparison with normal control, were analyzed.Results: In the noninfected rabbit, the stroma was densely innervated in the central area and as a consequence the sub-basal epithelial nerve bundles were shorter, and no vortex was found. The HSV-infected corneas showed nerve damage in both epithelial and stromal nerves. Corneas infected with ICP0 and gK deletion mutants showed mild to moderate damage, while those infected with 17Syn+ and McKrae strains were seriously damaged. In the eyes infected with ICP0 and gK deletion, there were reduced numbers of sub-basal nerve bundles, but most of the corneas retained a normal stromal network. Corneas infected with 17 Syn+ and McKrae displayed destroyed nerve structures and formation of a scar tissue in the central cornea, in which only a few nerve fibers could be detected.Conclusion: HSV-1 primary corneal infection seriously damages the corneal nerves, persisting for more than 4 months. Reduction of axonal transport (by gK deletion) or virus replication (by ICP0 deletion) significantly attenuated the nerve damage induced by the virus.