Infection of a human respiratory epithelial cell line with rhinovirus. Induction of cytokine release and modulation of susceptibility to infection by cytokine exposure.

Abstract

Rhinovirus infections cause over one third of all colds and are a contributing factor to exacerbations of asthma. To gain insights into the early biochemical events that occur in infected epithelial cells, we develop, for the first time, a model in which a pure respiratory epithelial cell population can be routinely infected by rhinovirus. Viral infection was confirmed by demonstrating that viral titers of supernatants and lysates from infected cell increased with time and by PCR. Infection by rhinovirus 14 was inhibited by homotypic antiserum and by antibodies to intercellular adhesion molecule-i (ICAM-1), the receptor for this virus. Susceptibility of epithelial cells to infection by rhinovirus 14 (but not rhinovirus 2, an ICAM-1 independent strain) can be increased by preexposure of cells to TNFa, whereas IFNy reduces susceptibility to infection by both rhinovirus strains. Rhinovirus infection per se does not markedly alter ICAM-1 expression on epithelial cells. Finally, we demonstrate that rhinovirus infection induced increased production of IL-8, IL-6, and GM-CSF from epithelial cells. Production of IL-8 correlated with viral replication during the first 24 h after infection. This model should provide useful insights into the pathogenesis of rhinovirus infections. (J. Clin. Invest. 1995. 96:549-557.)

Role of NF-κB in Cytokine Production Induced from Human Airway Epithelial Cells by Rhinovirus Infection

“…At a higher infectious dose (3 ϫ 10 4 TCID 50 ), partial inhibition was observed (470 -230 pg/ ml). These data are consistent with our earlier observation that very little virus is necessary to begin a productive infection (8).…”

“…HRV-16 was propagated in WI-38 cells as previously described (8). The HRV-16 stock generated in this manner was purified to remove ribosomes and soluble factors of WI-38 origin by centrifugation through sucrose, according to published methods (22).…”

“…Both the filtrate, which should contain the same concentration of most known cytokines as the original preparation, and the concentrated retentate were then compared with the original preparation for their ability to generate cytokines. Second, BEAS-2B cell cultures were preincubated with 20 g/ml of mouse-blocking mAb to human ICAM-1 (84H10; Coulter-Immunotech, Miami, FL), or with a class-matched control Ab, as previously described (8). Cells were then exposed to varying doses of HRV-16, and cytokine production was assessed.…”

“…Measurements of IL-8 were performed using a previously described ELISA sensitive to 30 pg/ml of cytokine (8). Levels of IL-6 were assayed using a commercial kit sensitive to 15 pg of IL-6/ml (Biosource International, Camarillo, CA), whereas GM-CSF was quantified using a commercial ELISA sensitive to 7.8 pg of GM-CSF/ml (R&D Systems, Minneapolis, MN).…”

“…These studies were undertaken to further explore the potential role of NF-B in rhinovirus-induced cytokine production from epithelial cells. Because of our previous data demonstrating that the BEAS-2B bronchial epithelial cell line responds to rhinovirus infection in a manner similar to primary epithelial cells with a rapid and robust generation of IL-8, IL-6, and GM-CSF (8,11), we chose to conduct our studies using this cell line. We used oligonucleotides corresponding to the NF-B binding sites in the promoters of the IL-6 and IL-8 genes, as well as the consensus sequence originally described from the Ig gene (20), to examine the time course of NF-B activation in cells infected with partially purified human rhinovirus (HRV) 3 -16, relative to the time course of cytokine mRNA induction.…”