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J. Biol. Chem., Vol. 277, Issue 33, 30227-30235, August 16, 2002
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From the Department of Medicine, Division of Pulmonary and Critical
Care Medicine, The Johns Hopkins University,
Baltimore, Maryland 21224
Sphingosine 1-phosphate (S1P), a potent bioactive
sphingolipid, has been implicated in many critical cellular events,
including a regulatory role in the pathogenesis of airway inflammation. We investigated the participation of S1P as an inflammatory mediator by
assessing interleukin-8 (IL-8) secretion and phospholipase D (PLD)
activation in human bronchial epithelial cells (Beas-2B). S1P1, S1P3, S1P4,
S1P5, and weak S1P2 receptors were detected in
Beas-2B and primary human bronchial epithelial cells. S1P stimulated a
rapid activation of PLD, which was nearly abolished by pertussis toxin
(PTX) treatment, consistent with S1P receptor/Gi protein coupling. S1P also markedly induced Beas-2B secretion of IL-8, a
powerful neutrophil chemoattractant and activator, in a PTX-sensitive manner. This S1P-mediated response was dependent on transcription as
indicated by a strong induction of IL-8 promoter-mediated luciferase activity in transfected Beas-2B cells and a complete inhibition by
actinomycin D. Beas-2B exposure to 1-butanol, which converts the
PLD-generated phosphatidic acid (PA) to phosphatidylbutanol by a
transphosphatidylation reaction, significantly attenuated the
S1P-induced IL-8 secretion, indicating the involvement of PLD-derived
PA in the signaling pathway. Inhibition of
12-O-tetradecanoyl-phorbol-13-acetate-stimulated IL-8
production by 1-butanol further strengthened this observation. Blocking
protein kinase C and Rho kinase also attenuated S1P-induced IL-8
secretion. Our data suggest that PLD-derived PA, protein kinase C, and
Rho are important signaling components in S1P-mediated IL-8 secretion
by human bronchial epithelial cells.
To whom correspondence should be addressed: The Johns Hopkins
Asthma and Allergy Center, Division of Pulmonary and Critical Care
Medicine, 5501 Hopkins Bayview Circle, Baltimore, MD 21224. Tel.:
410-550-7748; Fax: 410-550-2612; E-mail: vnatara1@jhmi.edu.
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