Influenza A virus infection of human middle ear cells in vitro

Craig A. Buchman, Nevis Fregien

Research output: Contribution to journalArticle

10 Scopus citations

Abstract

Hypothesis: Human-derived normal middle ear mucosal cells can be harvested and cultured and will support influenza A virus (INF A) infection. Study Design: Protocols for the collection and in vitro culture of middle ear mucosal cells were developed and used to investigate the effects of INF A infection as it relates to the pathogenesis of otitis media. Materials and Methods: Middle ear mucosa was harvested during surgeries that opened the normal middle ear. Middle ear mucosal cells were plated and grown in collagen-coated dishes. Cells were characterized before and after INF A exposure using phase-contrast and immunofluorescence microscopy as well as reverse transcriptase-polymerase chain reaction (RT-PCR) for cytokeratin 18 gene expression and INF A. Results: Primary cultures of human middle ear epithelial cells were established. Prolonged growth of middle ear cells yielded a second cell type that failed to stain for cytokeratin on immunofluorescence but continued to produce positive RT-PCR results on cytokeratin 18 analysis. After INF A exposure, cytological changes and immunofluorescence staining showed cellular infection. RT-PCR analysis using INF A-specific primers showed positive results for up to 72 hours after viral exposure. Conclusions: Primary cultures of human middle ear mucosal cells have been established. Two distinctly different cell culture systems have been developed: 1) middle ear epithelial cells and 2) either dedifferentiated epithelial cells or fibroblasts. Exposure of both cell types to INF A demonstrates that each can support cellular infection and viral replication. These models should be useful for studies of the pathogenesis of virus-mediated otitis media.

Original languageEnglish (US)
Pages (from-to)1739-1744
Number of pages6
JournalLaryngoscope
Volume110
Issue number10
DOIs
StatePublished - 2000

Keywords

  • Cell culture
  • Influenza virus
  • Middle ear epithelium
  • Otitis media

ASJC Scopus subject areas

  • Otorhinolaryngology

Fingerprint Dive into the research topics of 'Influenza A virus infection of human middle ear cells in vitro'. Together they form a unique fingerprint.

  • Cite this