Ocular Reactivation of MCMV after Immunosuppression of Latently Infected BALB/c Mice

Abstract

The purpose of this study was to identify the site(s) of MCMV latency and reactivation in the eye. Three months after supraciliary inoculation of 5 x 10(2) PFU of MCMV, BALB/c mice underwent immunosuppression with methylprednisolone and antibodies specific for CD4 T cells, CD8 T cells, and NK cells or with methylprednisolone alone. Control mice were infected but did not receive the immunosuppressants. After 2 or 3 weeks of immunosuppression, the mice were killed. Replicating virus and viral antigen were detected in the injected eyes, peripheral blood leukocytes (PBLs), and extraocular tissues by plaque assay and by staining for early antigen (EA) and beta-galactosidase (beta-gal), respectively. In latently infected, nonimmunosuppressed control mice, replicating-virus-and viral-antigen-positive cells were not detected in the injected eyes or extraocular tissues. After immunosuppression with methylprednisolone and antibodies, EA and beta-gal were detected, and replicating virus was recovered from the injected eye and from several extraocular sites, including liver, lungs, salivary glands, and kidneys. No virus was recovered from PBLs. beta-Gal- or EA-positive cells were observed in the RPE of most mice, and a few virus-infected cells were also observed in the nuclear layers and ganglion cells. Microscopic changes, including retinal folding and detachment, photoreceptor atrophy, macrophage infiltration, and a few EA-positive cytomegalic cells, were observed in the injected eye of immunosuppressed mice. After immunosuppression, MCMV reactivates in the injected eye and extraocular tissues, and RPE cells are the initial site of MCMV ocular reactivation in the eye. The timing of virus recovery from all sites suggests that MCMV observed in the injected eye is from in situ reactivation of virus and not from spread of virus from extraocular sites via infected PBLs.