Infection of cultured human tracheal epithelial cells by human parainfluenza virus types 2 and 3.
Despite growing information of the effects of human respiratory virus infection on airway physiology, little information is available on the mechanisms of pathology and pathophysiology in these infections. The human respiratory pathogens, parainfluenza virus types 2 and 3 (hPIV2, hPIV3, respectively), clinically cause laryngotracheobronchitis (infection of the large proximal airways). In order to examine the pathobiology of these viruses in airway cells of human origin, we exposed primary cultures of human tracheal epithelial cells. Primary cultures of human tracheal epithelial cells were readily infected by these agents: cells exposed to hPIV2 and hPIV3 expressed viral antigens (demonstrated by indirect immunofluorescence assay), produced infectious virus, and demonstrated cytopathic effects (including early syncytium formation). Peak viral titers of 2 x 10(7) plaque-forming units per milliliter were obtained, similar to titers from permissive CV-1 cells. Trypan blue staining and direct cell counts demonstrated no difference in the viability of the control and infected cells until the infected cells began to detach from the culture substrate. However, infected cells release significantly more LDH than control cells by 48 h following infection at a multiplicity of infection of 1 virus/target cell. This system provides a model for studying the effects of infection of the human tracheal epithelium by human respiratory viral pathogens without confounding interactions with other cell and tissue types.
Stark
J M
JM
Department of Pediatrics, Case Western Reserve, University School of Medicine, Cleveland, OH 44106.
Huang
Y T
YT
Carl
J
J
Davis
P B
PB
D000328:Adult; D002452:Cell Count; D002470:Cell Survival; D002478:Cells, Cultured; D004847:Epithelial Cells; D004848:Epithelium / Q000382:microbiology; D005455:Fluorescent Antibody Technique; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D010223:Parainfluenza Virus 2, Human / Q000472:pathogenicity / Q000502:physiology*; D010224:Parainfluenza Virus 3, Human / Q000472:pathogenicity / Q000502:physiology*; D013223:Statistics as Topic; D014132:Trachea / Q000382:microbiology*; D014343:Trypan Blue; D014776:Virus Cultivation
D007770:L-Lactate Dehydrogenase; D014343:Trypan Blue