|Knowles, Donald - Don|
Submitted to: Veterinary Microbiology
Publication Type: Peer reviewed journal
Publication Acceptance Date: 3/9/2010
Publication Date: 9/28/2010
Citation: Taus, N.S., Schneider, D.A., Oaks, J.L., Yan, H., Gailbreath, K.L., Knowles Jr, D.P., Li, H. 2010. Sheep (Ovis aries) airway epithelial cells support ovine herpesvirus 2 lytic replication in vivo. Veterinary Microbiology. 145(1-2):47-53. Interpretive Summary: Domestic sheep are the carriers of ovine herpesvirus 2 (OvHV-2), the virus that causes sheep-associated malignant catarrhal fever (MCF) in domestic cattle and American bison. We have previously shown that OvHV-2 first replicates in the lungs of experimentally infected sheep and that this initial lytic replication is well controlled by the immune system. Additionally, we have shown that the nasal cavities of naturally infected sheep are the sites of OvHV-2 lytic replication during intense virus shedding events. However, we did not identify the specific types of cells in which the virus was replicating. In this study we describe the development of an antibody that detects OvHV-2 protein. We used this antibody in combination with other antibodies that detect various cellular proteins to determine that epithelial cells in the lungs and nasal cavities of sheep are the specific cells that supported OvHV-2 lytic replication. This information will guide the next step in developing a cell culture system to grow OvHV-2.
Technical Abstract: In this study we describe the development of a monospecific, polyclonal rabbit antiserum directed against the ovine herpesvirus 2 (OvHV-2) major capsid protein and its use to detect lytically infected cells in domestic sheep (Ovis aries). Immunofluorescent labeling using monoclonal antibodies directed against cellular proteins and the OvHV-2 capsid-specific antiserum identified infected alveolar epithelial cells in formalin-fixed lung sections of experimentally infected sheep seven days post inoculation. The same technique was used to detect infected epithelial cells in cytospin preparations made from nasal swabs collected from naturally infected sheep during virus shedding episodes. This is the first reported identification of specific cell types that support OvHV-2 lytic infection in vivo and this information will be used to design experiments to test whether various epithelial cells can be used to propagate OvHV-2 in vitro.