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Debra A. Tristram, MD; Wesley Hicks, Jr, MD; Robert Hard, PhD

Arch Otolaryngol Head Neck Surg. 1998;124:777-783.

Background  A suitable model for respiratory syncytial virus (RSV) infection has yet to be developed.

Objective  To describe an in vitro model of human respiratory epithelium in primary cell culture linked with a computer microscope interface that allows evaluation and imaging of living RSV-infected respiratory epithelium.

Design  A descriptive, controlled study. Human bronchial cells were obtained from surgical samples by elastase dissociation and replated on collagen gel membranes. After 7 to 10 days, cells were brought to air interface. Baseline sampling of cell fluid for cytokine production by enzyme-linked immunosorbent assay (interleukin [IL] 1, IL-6, IL-8, and RANTES) and leukotriene C4 by radioimmunoassay was taken before treatment with RSV (n=30) or HEp-2 (human laryngeal carcinoma cells) control (n=25). Sampling was done at 4, 24, 72, and 120 hours thereafter. The infectious process was monitored with a microscope (Zeiss UEM, Carl Zeiss Inc, Thornwood, NY) equipped with a camera (Newvicon, Dage Corporation, Stamford, Conn). Images were either digitized using a computer (Macintosh Quadra 950, Apple Computers Inc, Cupertino, Calif) equipped with a digitizing board (Perceptics Corporation, Knoxville, Tenn) or were recorded on an SVHS videotape using a videocassette recorder (JVC, Elmwood Park, NJ).

Results  Respiratory syncytial virus induced profound effects on the ciliated cells: ciliostasis, clumping, and loss of cilia from live cells and sloughing of cells. Significant differences in the release of IL-6, IL-8, and RANTES (P<.03 for each cytokine) were noted in RSV-infected bronchial cultures by 24 hours with a peak at 72 hours. The IL- and leukotriene C4 were not altered by RSV infection in bronchial cells.

Conclusions  This model closely mirrors human RSV disease and affords a unique opportunity to study interepithelial cell interactions, cytokine responses from cells of different donors, and ciliary activity of live cells undergoing RSV infection.

From the Departments of Pediatrics (Dr Tristram), Surgery (Dr Hicks), and Anatomy (Dr Hard), State University of New York at Buffalo School of Medicine, Children's Hospital of Buffalo (Dr Tristram), and Roswell Park Cancer Institute (Dr Hicks), Buffalo, NY.

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