The Baker Institute for Animal Health
Email: firstname.lastname@example.org; Phone: 607-255-5624
Sponsor: Research Grants Program in Animal Health
Grant Number: N/A
Title: The Air-Liquid Canine Corneal Organ Culture System: A New Tool to Study the Pathogenesis of Acute Alphaherpesvirus-Induced Ocular Disease
Project Amount: $ 47,985
Project Period: 09/01/13-08/31/15
DESCRIPTION: Herpesvirus infection is an important cause of disease in dogs, cats, humans and many other animal species. Infection occurs via mucosal surfaces of the respiratory and genital tracts or via epithelial surfaces such as the cornea of the eye. Herpes simplex virus (HSV)-induced keratitis is the leading infectious cause of visual impairment and blindness in humans. Similarly, in companion animals, it is increasingly recognized that alphaherpesviruses are an important cause of ocular disease. Despite the clinical importance of herpesvirus-induced ocular disease, the underlying pathophysiology of this disease remains poorly understood, in part because of the lack of good in vitro models in which to study the cellular and viral factors involved in acute corneal infection. Canine herpesvirus type 1 (CHV1) causes a variety of diseases in dogs including mortality in puppies and acute ocular disease in adult dogs. In order to be able to develop rationally-based vaccines and novel antivirals for acute ocular herpesvirus infection in dogs, a comprehensive understanding of the pathogenic mechanisms and the viral factors involved during CHV1 infection in the eye is urgently needed. Such studies are hampered, however, as no in vitro models are available that would allow us to study viral replication and virus-host interaction of CHV1 in corneal epithelial cells or tissues.
We hypothesize that an air-liquid canine corneal organ culture system will support CHV1 infection and will serve as a valuable virus-natural host model system in which to study the pathogenesis of acute alphaherpesvirus-induced ocular disease. To test this hypothesis, following specific aims are proposed. In Aim 1, we will establish and validate the air-liquid canine corneal organ culture system for studying acute CHV1-induced corneal infection. In Aim 2, we will develop the necessary tools to create CHV1 mutant viruses, deleted in genes reported to be ocular virulence determinants. In Aim 3, we will examine the replication kinetics of CHV1 and evaluate the CHV1-induced corneal damage in our newly developed air-liquid canine corneal organ culture system. Hereby, clinical isolates as well as the mutant viruses of CHV1 will be studied.
Our preliminary data show that we can maintain canine corneas in culture for at least one week using this air-liquid system and that this system is susceptible for CHV1. We expect that CHV1 will replicate in our air liquid canine corneal organ culture system and we predict that we will identify differences in replication kinetics and/or corneal damage between clinically different CHV1 field isolates as well as CHV1 mutant viruses.
The research in this proposal will advance our knowledge on the mechanisms of acute CHV1-induced ocular disease. Specifically, the project will investigate the replication kinetics and corneal damage of acute CHV1 in a unique air-liquid canine corneal culture system. By developing novel tools for herpesvirus research and using those to study viral replication in a physiologically relevant and natural host model, we contend that we have a powerful tool to functionally unravel herpesvirus pathogenesis in the eye. We are convinced that a better understanding of the mechanisms by which alphaherpesviruses induce acute ocular disease will be critical in the rational design of safer and more efficacious vaccines, and in the identification of antiviral drugs that can be safely applied to the corneal epithelium.