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Infect. Immun., 07 1997, 2648-2655, Vol 65, No. 7
K Oishi, B Sar, A Wada, Y Hidaka, S Matsumoto, H Amano, F Sonoda, S Kobayashi, T Hirayama, T Nagatake and K Matsushima
Persistent infection with Pseudomonas aeruginosa increases interleukin- 8
(IL-8) levels and causes dense neutrophil infiltrations in the airway of
patients with chronic airway diseases. To investigate the role of P.
aeruginosa infection in IL-8 production in the airway of these patients, we
examined whether cell lysates of P. aeruginosa could cause IL-8 production
from human bronchial epithelial cells. Diluted sonicated supernatants of P.
aeruginosa (SSPA) with a mucoid or nonmucoid phenotype stimulated human
bronchial epithelial (BET-1A) cells to produce IL-8. In this study, we have
purified a 59-kDa heat- stable protein with IL-8-inducing activity from the
SSPA by sequential ion-exchange chromatography. The N-terminal sequence of
this purified protein completely matched a sequence at the N-terminal part
of the mature protein of nitrite reductase from P. aeruginosa. In addition,
immunoblotting with a polyclonal immunoglobulin G (IgG) against recombinant
Pseudomonas nitrite reductase demonstrated a specific binding to the
purified protein. Furthermore, the immunoprecipitates of the SSPA with a
polyclonal IgG against recombinant nitrite reductase induced a
twofold-higher IL-8 production in the BET-1A cell culture than did the
immunoprecipitates of the SSPA with a control IgG. These lines of evidence
confirmed that Pseudomonas nitrite reductase was responsible for IL-8
production in the BET-1A cells. The purified nitrite reductase induced
maximal expression of IL-8 mRNA in the BET-1A cells at 1 to 3 h after
stimulation, and the IL-8 mRNA expression declined by 8 h after
stimulation. New protein translation was not required for nitrite
reductase-mediated IL-8 mRNA expression in the BET- 1A cells. Nitrite
reductase stimulated the BET-1A cells, as well as human alveolar
macrophages, pulmonary fibroblasts, and neutrophils, to produce IL-8. In
contrast, nitrite reductase induced significant levels of tumor necrosis
factor alpha and IL-1beta protein only in human alveolar macrophages. These
data support the notion that nitrite reductase from P. aeruginosa induces
the production of inflammatory cytokines by respiratory cells and may
contribute to the pathogenesis of chronic airway diseases and persistent P.
aeruginosa infection.
Copyright © 1997, American Society for Microbiology
Nitrite reductase from Pseudomonas aeruginosa induces inflammatory cytokines in cultured respiratory cells
Department of Internal Medicine, Institute of Tropical Medicine, Nagasaki University, Sakamoto, Japan. oishi-k@net.nagasaki-u.ac.jp
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