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There is an intense infiltrate of CD4+ (blue) and CD8+ (orange) T cells and mixed populations of DCs (purple), macrophages (teal) and NK cells (olive). Vaccine and Infectious Diseases Division, Fred Hutchinson Cancer Research Center, Seattle, Washington, USA. HSV is continually and slowly released due to concurrent reactivation of HSV within multiple ganglia. The virological, clinical, and histopathological manifestations of acute and experimentally reactivated infections of eyes and trigeminal ganglia have been studied following intranasal infection of rabbits with herpes simplex virus type 1 (strain KOS-63). References1.Arao Y, Hatano A, Yamada M, Uno F, Nii S (1991) Reactivatable latency of three avirulent strains of herpes simplex virus type 1 after intranasal inoculation of mice. JavaScript is currently disabled, this site works much better if you enable JavaScript in your browser. Schematic diagram of the ROSA26R transgene and Cre expression cassette, and herpes simplex virus type 1 (HSV-1) Cre-expressing recombinant viruses. This journal is a member of and subscribes to the principles of the Committee on Publication Ethics. All animals shed virus in nasal secretions, but only three shed virus in tear film during the first 12 days of infection.

Periodically, epithelial cells become infected, leading to rapid viral spread, multiple infected cells, microulcer or ulcer formation (where high numbers of keratinocytes are lysed and virions are shed) and dense CD4+ and CD8+ T cell infiltration. No animal developed clinical or histological evidence of corneal or retinal ocular disease at any time after infection.
The location and function of macrophages, B cells and neutralizing antibodies have not been fully characterized. After containment of HSV at a single site, T cells persist at that location for several months in an immunosurveillance capacity. KOS-63 established trigeminal ganglionic latency; viral RNA, restricted to neuronal nuclei, was detected by in situ hybridization, and virus was recovered from co-cultivation cultures of nervous tissue, but not from cell-free homogenates. The delivery of the DNA recombinase Cre results in the reciprocal recombination between the loxP sites and the excision of the neo cassette, leaving behind lacZ under the control of the ubiquitous and constitutive ROSA promoter. HSV-2 reactivates at other sites where CD4+ and CD8+ lymphocyte density is lower, allowing for a genital tract shedding episode, which initiates in a different region.
Reactivation of latent trigeminal ganglionic infection was attempted by intravenous administration of cyclophosphamide, followed by dexamethasone 24 h later. Injection of the drugs failed to reactivate KOS-63 latency; no animal shed virus in nasal or ocular secretions, and no animal developed gross or microscopic corneal lesions.

The HSV-1 genome is depicted at the top, and insertions in the nonessential TK and UL43 loci are shown below. In addition, viral antigens were not detected by immunofluorescence microscopy in ganglia from rabbits subjected to the drug protocol, and virus was only recovered from ganglia by in vitro co-cultivation reactivation techniques. The HCMV MIEP is represented by the striped box, the SV40 polyadenylation and termination sequences by the shaded box, and the Cre coding region by the open box. The failure of KOS-63 to reactivate was not due to an inherent failure of populate and infect the ganglion, because the virus did not reactivate from ganglia that contained many latently infected cells. These studies demonstrate that, although KOS-63 is neuroinvasive and capable of establishing latency, it is virtually non-virulent for the eye, and cannot be reactivated by a systemic immunosuppressive trigger known to reactivate other HSV-1 strains. At the indicated times, duplicate cultures were assayed for virus content by standard PFU assays.

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