title: Use of a feline respiratory epithelial cell culture system grown at the air-liquid interface to characterize the innate immune response following feline herpesvirus 1 infection.
abstract: Infection with feline herpesvirus-1 (FHV-1) accounts for 50% of viral upper respiratory diseases in domestic cats and is a significant cause of ocular diseases. Despite the clinical significance and high prevalence of FHV-1 infection, currently available vaccines cannot completely protect cats from infection and lifelong latency. FHV-1 infects via the mucous membranes and replicates in respiratory epithelial cells, but very little is known about the early innate immunity at this site. To address questions about immunity to FHV-1, feline respiratory epithelial cells cultured at air-liquid interface (ALI-FRECs) were established by collecting respiratory tracts from 6 healthy cats after euthanasia. Cells were isolated, cultured and characterized histologically and immunologically before infection with FHV-1. The expression of Toll-like receptors (TLRs), cytokine and chemokine responses were measured by real time PCR. ALI-FRECs morphologically resembled the natural airways of cats with multilayered columnar epithelial cells and cilia. Immunological properties of the natural airways were maintained in ALI-FRECs, as evidenced by the expression of TLRs, cytokines, chemokines, interferons, beta-defensins, and other regulatory genes. Furthermore, ALI-FRECs were able to support infection and replication of FHV-1, as well as modulate transcriptional regulation of various immune genes in response to infection. IL-1β and TNFα were increased in ALI-FRECs by 24hpi, whereas expression levels of IFN-α and TLR9 were not increased until 36hpi. In contrast, TLR3, GM-CSF and TGF-1β expression was down-regulated at 36hpi. The data presented show the development of a system ideal for investigating the molecular pathogenesis and immunity of FHV-1 or other respiratory pathogens.
authors.affiliation: Department of Pathobiology and Diagnostic Investigation, College of Veterinary Medicine, 784 Wilson Road, East Lansing, MI 48824, USA. Electronic address: rknelli@msu.edu.
authors.forename: Rahul K
authors.initials: RK
authors.lastname: Nelli
authors.affiliation: Diagnostic Center for Population and Animal Health, 4125 Beaumont Road, Building 0215, Lansing, MI 48910, USA. Electronic address: maes@dcpah.msu.edu.
authors.forename: Roger
authors.initials: R
authors.lastname: Maes
authors.affiliation: Diagnostic Center for Population and Animal Health, 4125 Beaumont Road, Building 0215, Lansing, MI 48910, USA. Electronic address: kiupel@dcpah.msu.edu.
authors.forename: Matti
authors.initials: M
authors.lastname: Kiupel
authors.affiliation: Department of Pathobiology and Diagnostic Investigation, College of Veterinary Medicine, 784 Wilson Road, East Lansing, MI 48824, USA. Electronic address: husseygi@msu.edu.
authors.forename: Gisela Soboll
authors.initials: GS
authors.lastname: Hussey
mesh_terms: D000818:Animals; D002371:Cat Diseases / Q000276:immunology* / Q000378:metabolism / Q000821:virology*; D002415:Cats; D002460:Cell Line; D002478:Cells, Cultured; D018925:Chemokines / Q000235:genetics / Q000378:metabolism; D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D004847:Epithelial Cells; D015870:Gene Expression; D007113:Immunity, Innate / Q000235:genetics; D018836:Inflammation Mediators / Q000378:metabolism; D051192:Receptors, Pattern Recognition / Q000235:genetics / Q000378:metabolism; D051193:Toll-Like Receptors / Q000235:genetics / Q000378:metabolism; D018140:Varicellovirus / Q000502:physiology*
chemical_list: D018925:Chemokines; D016207:Cytokines; D018836:Inflammation Mediators; D051192:Receptors, Pattern Recognition; D051193:Toll-Like Receptors
keywords: Air–liquid interface cell culture; Cytokines; Feline herpesvirus-1; Innate immunity; Respiratory epithelium; Toll-like receptors