Ex vivo model of herpes simplex virus type I dendritic and geographic keratitis using a corneal active storage machine

Abstract

Background Herpetic keratitis (HK) models using whole human corneas are essential for studying virushost relationships, because of high species specificity and the role of interactions between corneal cell populations that cell culture cannot reproduce. Nevertheless, the two current corneal storage methods (hypothermia and organ culture (OC)) do not preserve corneas in good physiological condition, as they are characterized by epithelial abrasion, stromal oedema, and excessive endothelial mortality. Methods To rehabilitate human corneas intended for scientific use, we used an active storage machine (ASM) that restores two physiological parameters that are essential for corneal homeostasis: intraocular pressure and storage medium renewal (21mmHg and 2.6 μL/min, respectively). ASM storage regenerates a normal multilayer epithelium in 2 weeks. We infected six pairs of corneas unsuitable for graft by inoculating the epithelium with herpes simplex virus type 1 (HSV-1), and compared each ASM-stored cornea with the other cornea stored in the same medium using the conventional OC method. Results Only corneas in the ASM developed a dendritic (n = 3) or geographic (n = 2) epithelial ulcer reproducing typical HSV-1-induced clinical lesions. Corneas in OC showed only extensive desquamations. None of the uninfected controls showed epithelial damage. Histology,

Herpes simplex keratitis: A brief clinical overview

“…The quest for better diagnosis, prevention, and management of HSK was uppermost in the minds of sampled participants in a paper by Liu et al [ 461 ]. Novel corneal active storage mediums enable better study and research of ex-vivo disease patterns in herpetic keratitis[ 462 ]. In the future, target extraocular (maxillary) vaccination to inhibit ocular herpes simplex reactivation may improve the epidemiology of the disease[ 463 ].…”

Design and validation of a custom-made system to measure transepithelial electrical impedance in human corneas preserved in active storage machine

“…Most interestingly, the infection of human corneas stored in ASM by herpes simplex virus 1 lead to the development of characteristic dendritic lesions and cytopathogenic effect highly comparable to the lesions developed by patients. These lesions were not described in existing ex-vivo models, and were directly related to the presence of a mature multilayer epithelium ( Courrier et al, 2020 ). Taken together, these results support the pertinence of human corneas storage in ASM to develop alternative physiopathological models.…”

Cluster of Symptomatic Graft-to-Host Transmission of Herpes Simplex Virus Type 1 in an Endothelial Keratoplasty Setting

“…Second, HSV-1 replication in donor corneas during the culture period at the eye bank can occur without cellular changes in the graft. 37,38 Thus, microscopic examination performed before delivery by the eye bank may miss an ongoing HSV-1 infection in the graft. Therefore, we propose to perform HSV-1 PCRs on donor cornea culture media before its scheduled release for transplantation, and if positive, reject the donor graft.…”