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Department of Medicine, University of Iowa College of Medicine and Veterans Administration Medical Center, Iowa City, IA 52242
Respiratory syncytial virus (RSV) is an important respiratory
pathogen that preferentially infects epithelial cells in the airway and
causes a local inflammatory response. Very little is known about the
second messenger pathways involved in this response. To characterize
some of the acute response pathways involved in RSV infection, we used
cultured human epithelial cells (A549) and optimal tissue
culture-infective doses (TCID50) of RSV. We have previously
shown that RSV-induced IL-8 release is linked to activation of the
extracellular signal-related kinase (ERK) mitogen-activated protein
kinase pathway. In this study, we evaluated the upstream events
involved in ERK activation by RSV. RSV activated ERK at two time
points, an early time point consistent with viral binding and a later
sustained activation consistent with viral replication. We next
evaluated the role of protein kinase C (PKC) isoforms in RSV-induced
ERK kinase activity. We found that A549 cells contain the
Ca2+-dependent isoforms 
and 
1, and the
Ca2+-independent isoforms 
, 
, 
, µ, 
, and 
.
Western analysis showed that RSV caused no change in the amounts of
these isoforms. However, kinase activity assays demonstrated activation
of isoform within 10 min of infection, followed by a sustained
activation of isoforms 1, , , and µ 24–48 h postinfection.
A cell-permeable peptide inhibitor specific for the isoform
decreased early ERK kinase activation by RSV. Down-regulation of the
other PKC isoforms with PMA blocked the late sustained activation of
ERK by RSV. These studies suggest that RSV activates multiple PKC
isoforms with subsequent downstream activation of ERK
kinase.
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