Respiratory syncytial virus and human bronchial epithelium.
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.


METHODS
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] 1beta, 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-beta 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.
Tristram
D A
DA
Department of Pediatrics, State University of New York at Buffalo School of Medicine, Children's Hospital of Buffalo, 14222, USA. dtristra@acsu.buffalo.edu
Hicks
W
W
Hard
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R
D001980:Bronchi / Q000821:virology*; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism*; D004848:Epithelium / Q000821:virology; D006801:Humans; D008954:Models, Biological; D018357:Respiratory Syncytial Virus Infections / Q000378:metabolism*; D018113:Respiratory Syncytial Virus, Human / Q000302:isolation & purification / Q000472:pathogenicity*
D016207:Cytokines